The Magazine from Carl Zeiss in Memory of Ernst Abbe

Home , Anton Dohrn, Apochromat, Carl Zeiss, Carl Zeiss AG, Ernst Abbe, Siegfried Czapski

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In Memory of Ernst Abbe Inhalt_01_Editorial_E.qxd 15.08.2005 9:07 Uhr Seite 2

Contents

Editorial Formulas for Success. . . ❚ Dieter Brocksch 3

In Memory of Ernst Abbe Ernst Abbe 4 Microscope Lenses 8 Numerical Aperture, Immersion and Useful Magnification ❚ Rainer Danz 12 Highlights from the History of Immersion Objectives 16 From the History of Microscopy: Abbe’s Diffraction Experiments ❚ Heinz Gundlach 18 The Science of Light 24 Stazione Zoologica Anton Dohrn, Naples, Italy 26 Felix Anton Dohrn 29 The Hall of Frescoes ❚ Christiane Groeben 30 Bella Napoli 31

From Users The Zebra Fish as a Model Organism for Developmental Biology 32 SPIM – A New Microscope Procedure 34 The Scourge of Back Pain – Treatment Methods and Innovations 38 ZEISS in the Center for Book Preservation ❚ Manfred Schindler 42

Across the Globe Carl Zeiss Archive Aids Ghanaian Project ❚ Peter Gluchi 46

Prizes and Awards 100 Years of Brock & Michelsen 48 Award for NaT Working Project 49

Product Report Digital Pathology: MIRAX SCAN 50 UHRTEM 50 Superlux™ Eye Xenon Illumination 50 Carl Zeiss Optics in Nokia Mobile Phones 51

Masthead 51

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Editorial

Formulas for Success...

Formulas describe the functions and processes of what It clearly and concisely describes the resolution of optical happens in the world and our lives. It is often the small, instruments using the visible spectrum of light and con- insignificant formulas in particular that play a decisive tributed to the improvement of optical devices. role in what we know and in the functionality of modern instruments and examination methods.

for the large...

The fiftieth anniversary of the death of Albert Einstein (1879-1955) also marks the centennial of his theory of relativity; a theory that revolutionized perceptions, made the processes of life more understandable and began to explain the dimensions of time and space. The short for- Scientist mula, E=m·c2, expresses the infinite complexity of our Entrepreneur world. Einstein had contact with Zeiss throughout the Social Reformer course of his scientific activities. In 1925 he wrote to the company Anschütz in Kiel about producing a gyrocom- The same year also saw the passing of another German pass: “The difficulties of manufacturing are so great – microscope manufacturer with connections to Abbe and accuracies of 10-4 have to be achieved – that Zeiss is Zeiss: Rudolf Winkel (1827-1905). During Abbe’s times, currently the only company capable of meeting the re- good microscopes were also built at Winkel’s workshop quirements.“ which was founded in Göttingen in 1857. Abbe visited Winkel’s workshop while he was a student in Göttingen. His visit in 1894 led to closer cooperation. In 1911, Zeiss became Winkel’s chief partner. In October 1957, the firm R. Winkel GmbH became part of the Carl Zeiss Foundation. The same year that Abbe died, Robert Koch (1843- 1919) received the Nobel Prize for Medicine for his examinations and discoveries while researching tuberculosis. In 1878, Robert Koch used the Abbe oil immersion system for the first time and was impressed by the “quantum leap“ made by the “Carl Zeiss Optical Workshop using Professor Abbe’s ingenious advice.“ In 1904, Carl Zeiss management presented Robert Koch with the 1000th 1/12 objective lens for homogenous oil immersion. Many articles in this issue are dedicated to Ernst Abbe ... and small things in life. and his times. We reflect on what Ernst Abbe meant to Carl Zeiss and what he did for optics, and we take a spe- 2005 also marks the 100th anniversary of the death of cial look at developments that were and continue to be Ernst Abbes (1840-1905). Numerous events throughout significantly influenced by Ernst Abbe and his scientific 2005 honor his many great achievements. His extensive results. This is emphasized by the image on the cover examinations within the scope of his activities at Carl pages: an historical tribute to the more than 150 years of Zeiss’ optical workshop also resulted in the formula for optical development with a focus on microscopy. the resolution of a microscope: d = 2n sin July 2005

Dr. Dieter Brocksch

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In Memory of Ernst Abbe

1857

Education and (1857-1859) and Göttingen (1859- cine and Science, in which he gave a early years 1861). Abbe completed his studies by total of 45 lectures in the period to obtaining his doctorate in Göttingen 1895. From 1866, he was a freelance Ernst Abbe was born in Eisenach on on the subject “Experiential substan- scientist with the court and university January 23, 1840, as the son of tiation of the theorem of equivalence mechanic Carl Zeiss in Jena. In 1870 master spinner and subsequent fac- between heat and mechanical ener- Abbe formulated the famous sine tory attendant Georg Adam Abbe gy“. He subsequently worked for two condition subsequently named after and his wife Christina. He attended years as a teacher at the Physics him, a condition that must be met by elementary school from 1846 to Association in Frankfurt/Main. any spherically corrected lens if it is 1850, after which he was a student also to be free from coma in at the local high school in Eisenach. Family and the neighborhood of the lens axis in He finished his school education in science microscope image formation. In the 1857 and graduated with above- same year he became an extraor- average grades. In the period to Abbe’s mother died early in his life: dinary professor at the University of 1861 he studied mathematics and July 14, 1857. During his studies, his Jena. On September 24, 1871, he physics at the Universities of Jena father married for the second time, to married Elise Snell, the daughter of lec- the widow Eva Margarethe Liebetrau turer Prof. Snell, a mathematics and on November 11, 1859. After his physics lecturer at the University of time in Frankfurt, Abbe joined the Jena. Three years later, the first child, In 2005, the book titled “Ernst Mathematical Association in Jena in his daughter Paula, was born. His fa- Abbe – Scientist, Entrepreneur 1863. In the same year, he obtained ther died on August 18, 1874. Abbe and Social Reformer” was his post-doctoral qualification as a was director of Jena Observatory published by Bussert & lecturer with his paper on “the laws from 1877 to 1900. On May 1, 1878, Stadeler, Jena Quedlinburg in the distribution of errors in obser- he was appointed as an honorary to mark the 100th anniversary vation series“. He taught as a private member of the London Royal Micro- of Ernst Abbe’s death. mathematics and physics lecturer at scopical Society. In June of the same the University of Jena. year, he became an ordinary hon- [ISBN 3-932906-57-8] In 1863 Abbe also became a mem- orary professor at the University of ber of the Jena Association of Medi- Jena. He was awarded the title of Dr.

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1863 1870 1875 1880

med. h. c. by the University of Halle allowed the implementation of high for Otto Schott in Jena. The following in 1883 and the title Dr. jur. h. c. by quality standards. The corporate or- year saw the signing of the new part- the University of Jena in 1886. In ganization was successfully focused nership agreement in which Abbe 1900 Abbe became a corresponding on growth by the clear allocation of became an active partner together member of the Imperial Austrian responsibilities for scientific, technical with Carl and Roderich Zeiss. Academy of Science in Vienna. In and commercial staff. In 1884 the Glastechnische Labo- 1901 he was appointed as an hon- From 1872, all ZEISS microscopes ratorium Schott & Gen. (later to be- orary member of the Saxon Academy were built in line with Abbe’s calcula- come Jenaer Glaswerk Schott & of Science and of the Academy of tions. Three years later, in 1875, Gen.) was founded by Otto Schott, Science in Göttingen. Abbe became a dormant partner in Ernst Abbe, Carl Zeiss and Roderich the Optical Workshop of Carl Zeiss. Zeiss. Abbe as an Abbe pledged not to increase his ac- In search of calcium fluoride for entrepreneur ademic activity beyond the current optical applications, Abbe traveled to measure and not to accept a profes- Oltscherenalp in Switzerland for the The process of integrating science sorship in Jena or elsewhere. One first time in 1886. After the death of into industry already started in the year later, he traveled to London to Carl Zeiss on December 3, 1888, 1860s. In addition to Carl Zeiss, attend the international exposition of Abbe became the sole owner of the Siemens and Bayer were also pio- scientific instruments on behalf of Zeiss works in 1889. At the same neers of this development. By hiring the Prussian Department of Educa- time, he discontinued his teaching scientific staff, Ernst Abbe was a tion. In 1878, due to his obligations activities at the University of Jena. decisive driving force behind this at Zeiss, he turned down the offer of From 1890 onwards, Abbe expanded process at Zeiss: the integration of a post as professor in Berlin instigat- the product spectrum on an ongoing R&D into the company was an impor- ed by Hermann von Helmholtz. He basis: measuring instruments (1890), tant step toward technology leader- also declined the offer of an ordinary camera lenses (1890), binoculars ship. The training of capable employ- professorship in Jena. (1894), astronomical instruments ees and successors also played a He initially came into contact with (1897) and photogrammetric instru- significant role in the entrepreneurial Dr. Otto Schott in 1879. Their collab- ments (1901). As a result, the num- and commercial areas. Competent oration began one year later. In 1882 ber of employees rose to over 2000 staff and constant quality control a private glass laboratory was set up by 1905.

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special

Global Player

As many as 100 years ago, Carl Zeiss was already what would now be called a global player: the first sales branches were founded in London (1894), Vienna (1902) and St. Petersburg (1903). Today, the company has 15 production facilities in Germany, USA, Hungary, Switzerland, Mexico, Belarus and China as well as 35 sales organizations and 100 agencies across the globe. 1888 1901

Abbe’s persistence in his endeavors As a reformer, Abbe was far ahead of the constitution also reflected Abbe’s to also provide other manufacturers his times with his socio-political social commitment. For example, a with new types of optical glass was ideas. In 1889, in order to safeguard council was set up to represent the of great help to the German optical the existence of the enterprises Carl interests of employees. Although this industry. He was skeptical about the Zeiss and SCHOTT irrespective of per- could not be seen as codetermination patenting of products, which he sonal ownership interests, Abbe set in the modern sense of the term, it saw as an obstruction to scientific up the Carl Zeiss Foundation which did entitle the representatives to progress in general. Not until compet- he made the sole owner of the Zeiss voice their opinion in all matters con- itive pressure made it unavoidable did works and partial owner of the cerning the enterprise. Paid vacation, the patenting of camera lenses and SCHOTT works in 1891. In the same profit sharing, a documented entitle- binoculars begin. However, his early year, Abbe transferred his industrial ment to pension payments, contin- pioneering work remained accessible assets to the Carl Zeiss Foundation ued pay in the event of illness and, for general use. With the aid of With the appropriate compensation, from 1900, the eight-hour working Abbe’s comparator principle, instru- Roderich Zeiss also transferred his day were further social milestones. ments for the highly accurate meas- shares to the Foundation, making it This all made the Foundation enter- urement of workpieces were pro- the sole owner of the firm Carl Zeiss prises Carl Zeiss and SCHOTT fore- duced. These were important aids for and partial owner (sole owner from runners of modern social legislation. instrument construction in Germany. 1919) of Jena Glaswerk Schott & Tolerance was central to Ernst Abbe retired in 1903. To mark this Gen. Until 1903, Abbe was the au- Abbe’s basic philosophy of life. Al- occasion, a torchlight procession took thorized representative and one of though he was certainly not a social place with 1500 employees of the the three directors of the Carl Zeiss democrat, it was important to him Foundation companies. Two years Foundation. The corporate statute of that this political party was able to later, Ernst Abbe died on January 14, the Foundation came into force in evolve and develop freely. He was 1905, after a long, serious illness. 1896. The first supplementary statute also vehemently against racism, a followed just four years later. phenomenon which was already Abbe the social With his corporate statute of prevalent during his times. He en- reformer and the 1896, Abbe gave the enterprise a sured that no-one at Carl Zeiss suf- Carl Zeiss Foundation unique constitution. In addition to its fered in any way due to their origin, exceptionally progressive stipulations religion or political affiliation. This Many companies introduced their concerning corporate management attitude was reflected in the fact social policy in the late 19th century. and legally anchored labor relations, that two of his closest management

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1905

special colleagues, Siegfried Czapski and Products for Rudolf Straubel, were Jewish citizens. Science

Promoter of science Shortly after the development of and research the new microscopes, decisive breakthroughs were made in the Abbe always attached great impor- field of bacteriology. In 1904 tance to the promotion not only of Robert Koch wrote: “I owe a large science and research, but also of cul- proportion of the success I have ture. As a private citizen, Abbe sup- achieved for science to your excel- ported the university with anony- lent microscopes”. In the decades mous donations. The university and before the First World War medical the city of Jena were both sponsored research in Germany had a world by the Carl Zeiss Foundation after its standing that was paralleled only creation. As early as 1886, the pro- by the reputation enjoyed by motion of science and research be- ZEISS instruments. Emil Behring in gan with the secret “endowment the field of serology or Paul Ehrlich fund for scientific purposes“. in the field of chemotherapy are only two examples of many. Need- less to say, their success was not attributable to their instruments alone, but the microscopes did play an important role. The firm Carl Zeiss also created products for the field of chemistry, some of which were customized solutions: the gas interferometer for Fritz Haber, for example.

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Microscope Objectives

Figs 1-3: Early immerson objectives.

1 2 3

The quality displayed by early duced a new lens combination – it The Wollaston prism of the DIC microscope objectives was usually consisted of two plano-convex lenses microscopy technique is named very modest. The images were with a stop in the middle. Sir David after him. Wollaston discovered slightly blurred. Producing a mi- Brewster’s (1781-1868) idea of man- the elements palladium and croscope objective required a lot ufacturing objective lenses from dia- rhodium and was the first scien- of hard, intricate work until well monds was implemented in 1824 by tist to report about the dark into the second half of the 19th Andrew Pritchard. Brewster recom- lines in the solar spectrum. century. The standard complex mended using oil immersion to se- His way of viewing the geo- trial and error process needed to cure achromatism in 1813. In 1816, metric arrangement of atoms construct the optical systems was Joseph von Fraunhofer (1770-1841) led him to crystallography and extremely time consuming and produced the first achromatic lens William Hyde Wollaston (1766-1828), to the invention of the modern thus expensive. that could be used in microscopy. In English physicist, chemist goniometer for the angular 1823, Paris based physicist Selligue and philospher. measurement of crystal sur- combined up to four achromatic faces. In 1806 he invented the cemented elements into one objec- Camera lucida, an optical aid tive – this was the breakthrough in for perspective drawing that Sometime around 1810, Joseph Jack- the manufacture of achromatic mi- allows the observation of an son Lister (1786-1869) referred to croscope objectives with high resolu- object on a drawing surface. the connection between the angular tion. In its basic structure, the aperture of the objective and the In an age of increasing mecha- Camera lucida is a glass prism attainable resolution for the first nization and the beginning of indus- ao with two reflecting surfaces time. Two years later, William Hyde trial manufacture, Carl Zeiss quickly inclined at an agle of 135° that Wollaston (1766-1828) improved the recognized the link between theory o generates the image of a scene optics of the simple microscope: and practice, between science and at right angles to the eye of using Wollaston doublets, he intro- production required to effectively the observer.

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Fig. 4: d d Positive lens element. f S1 S2 f f f Fig.5: Diagram showing image generation.

Fig. 6: R 2 Negative lens element. R R R1 1 2 Fig. 7: 4 5 6 Chromatic aberration.

Fig. 8: Spherical aberration. manufacture powerful instruments which are required for efficient proof new types of glass. This led Abbe that would be able to stand up to duction of objectives with consistent- to bring young glass maker Otto the pressures of competition. Zeiss ly high quality, finally enabled Abbe Schott to Jena in 1882. Two years looked hard for a solution. The initial to produce objectives on a scientific later, Abbe and Zeiss became part- attempts to compute microscope ob- basis. He expanded the division of ners of the newly founded Glas- jectives between 1850 and 1854 by labor and specialization of employees technische Laboratorium Schott & himself and his friend, mathematician that Zeiss had started in the 1850s. Genossen. The new Schott glass ma- Friedrich Wilhelm Barfuss, however, Measuring machines and testing in- terials enabled Abbe to construct the did not return any noteworthy re- struments such as a thickness gages, apochromatic objective – the most sults. refractometers, spectrometers and powerful microscope objectives of the In 1866, Carl Zeiss approached a apertometer later entered volume late 19th century – in 1886. young, private tutor, Ernst Abbe, and production. requested help in developing im- In his early works, Abbe already proved microscope objectives. Over recognized that microscope objec- the next few years, Abbe developed tives would only be able to achieve the new theory of microscope image their full performance with the help formation which is based on wave optics (theory of diffraction) and was published in 1873. During this time, the sine condition for imaging, which defines the resolution limits of a microscope, was also formulated. Using the new theory, Abbe calculated several new microscope objectives. The construction of measuring devices 7 8

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Fig. 9: Natural (left) and artificial fluorite.

Reason for Abbe’s …The mineral emerged from trip to Switzerland oblivion again in the summer of 1886. When visiting mineral inspec- 16 February 1889, lecture by Dr. tor B. Wappler in Freiberg (Saxony) Edmund von Fellenberg during a during his search for water-clear fluo- meeting of the Natural Research So- rite, Dr. Abbe, professor of physics at ciety in Bern, Switzerland: About the the University of Jena, had seen Fluorite of Oltschenalp and its Techni- pieces of this material which Wappler cal Utilization. “An historical, scientif- had received from me in exchange ic memorandum for later times”. for minerals from Saxony many years …In our Alps calcium fluoride or before. 9 fluorite is no rarity and is quite fre- …Wappler indicated that he had quently found in the area of pro- received the pieces from me and togine (gneiss granite), the various correctly stated that they had been types of gneiss and crystalline slate found in the lower Haslithal in the and sometimes with excellent color- canton of Bern in Switzerland. Act- ing and interesting crystal shapes… ing upon this information, Professor In 1830, on a scree slope at the Abbe traveled immediately to foot of the Oltschikopf (2235 m) on Switzerland to visit me. He showed the Oltscheren mountain pasture, me a piece of transparent fluorite some Alpine enthusiasts discovered and asked me whether I could tell fragments of a shining, spathic him where this mineral could be mineral of outstanding transparency found in Switzerland. that they naturally thought to be rock crystal.… Source: Mitt. Naturf. Ges. Bern (1889) p.202-219

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Calcium fluoride or

fluorite (CaF2)

Fluorite is a mineral of the halogeni- des class. It is not only a popular gem, but also an important raw material for the production of hydroflu- oric acid, fluorine and fluxing agents (e.g. for the manufacture of alumi- num) and for the etching of glass. Clear crystals are used as lens elements for optical instruments. Nowa- days, artificially produced fluorite is used in optics. The name fluorite co- Fig. 10: mes from the Latin word fluere (to The Alpine house in Bielen (Bühlen) on the Oltscheren flow) and resulted from the use of Alps in 1889. Ernst Abbe can the mineral as a fluxing agent in the be easily recognized in the extraction of metals. Fluorite displays enlarged section produced by Trinkler in 1930 using the the colors purple, blue, green, yellow, best Carl Zeiss copying colorless, brown, pink, black and red- lenses available at that time. dish orange. At the end of the 19th century Ernst Abbe was the first person to use natural fluorite crystals in the construction of microscope objective lenses in order to enhance chromatic correction.

Oltscheren mountain pasture (1623 m)

Hut no. 86 with the inscription “B 1873 H*: edifice on a solid foundation, with a kitchen-cum-living room since 1996; saddle roof with …, three-room living area facing NE on the first floor (two rooms in the front, behind them a now unused kitchen and behind this a store and barn. Beside these a milking installa- tion with 5 stands. The hut belonged to the Zeiss Works in Jena during the period when the company was mining for fluorite.

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Numerical Aperture, Immersion and Use

Fig. 1: Causal connection Illustration of the invariance between numerical of the numerical aperture with regard to refraction at a aperture and plano-parallel glass plate resolution (e.g. a coverslip).

n1 = 1.518 , The key parameter of a microscope is 1 = 40° , known to be its ability to resolve n2 = 1.0 , minute object details, and not its = 77.4° , 2 magnification. To define the resolv- n1 sin 1 = n2 sin 2 ing power and its reciprocal value, the limit of resolution, Ernst Abbe Fig. 2: Large-aperture dry objective: coined the term numerical aperture the sine condition has been (apertura [lat.] = opening, numerical 2 3 met (explained in the text). aperture = dimensionless aperture). Fig. 3: The numerical aperture is the prod- Same objective type: the sine uct of the refractive indexOR and the Readers interested in mathematics object points (the “weights” on the condition has not been met; sine of half the angular aperture in might be surprised to see that Abbe dumbbell-shaped object 2∆y in off-axis image points display min pronounced coma the object space, and has one deci- used the sine instead of the tangent Fig. 4) must measure at least (explained in the text). sive advantage over the sole use of of half the angular aperture, as re- the parameter “angular aperture = quired by Gaussian image formation, (Figures 2 and 3 by courtesy of M. Matthä, Göttingen, 2 ”: its behavior is not changed by for example. The latter, however, is (3a) Germany). refraction at plano-parallel surfaces only concerned with very narrow ray 1.22 · (e. g. coverslips). pencils, i. e. the sines and tangents sin2 = min d Snell’s law of sines of the angular apertures are inter- changeable. After initial failures, since only then can the diffraction Abbe quickly realized that a very maximum of the one object point (1) specific condition must be met for coincide with the diffraction mini- microscopic imaging, namely the sine mum of the other, and hence both sin 1 n2 condition: if surface-elements are to object points can only just be seen = sin 2 n1 be imaged without error by means of separately (diffraction at a circular widely opened ray pencils, the ratio pinhole or lens edge; the number allows easy proof of this invariance between the numerical apertures on 1.22 is connected with the zero (Fig. 1): the object and image sides must point of the Bessel function). equal the lateral magnification, and thus must be constant. If this condi- In the microscope, however, the ∆ (2) tion has been met and the spherical resolution limit 2 ymin (longitudinal aberration1) (aberratio [Lat.] = going dimension) and not the angular reso- 1) (R.W. Pohl aptly n1 · sin 1 = n2 · sin 2 = ... = ni sin i astray) corrected, the image is descri- lution is of prime interest. R. W. Pohl described spherical bed as “aplanatic” ( – provides an amazingly simple deriva- aberration as “…poor combination [Gr.] = not going astray); off-axis tion of the microscopic resolution of axially symmetrical object points are imaged without co- limit from the sine condition: light bundles with a ma. Fig. 2 shows the 3-dimensional, In accordance with Fig. 4, formula wide opening...”) almost error-free diffraction image of (3a) can also be written as follows: an illuminated off-axis point (star test) that was captured with an objective meeting the sine condition. If (3b) the sine condition is not met, howev- 2 2 ∆ er, the image cannot be aplanatic, 2 y’min 1.22 · n2 = 1.0 (air) = 2 1 but displays pronounced coma (Fig. 3). b d Coverslip, There is a fundamental relationship According to Fig. 4, the numerical n1 = 1.518 between the sine condition and the aperture on the image side can be Object(point) of OP refractive index n resolving power: the angular separa- represented as follows: O 1 tion 2 min of two separately visible

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ful Magnification

(4) What should be done, however, if d a very specific wavelength or white 2 2’ n · sin ’= Lens diameter d BR 2b light must be used and if, with nOR = 1.0, the maximum value of 0.95, for 2 2’ (In general, the image space is filled example, has already been allocated with air, i.e. nBR = 1.0) to the dry aperture? In such cases, immersion objectives are used, i. e. 2 2’ When the sine condition is taken into objectives whose front lens immerses account, (immergere [Lat.]) into a liquid, the ab optical data of which has been in- 4 cluded in the objective’s computa- (5) tion. In the special case of homogeneous immersion, the refractive in- Objective front lens ∆ (e. g. BK 7, nOR · sin 2 y’ dices of the immersion liquid n2 and = ∆ = const. n3 = 1.518) nBR · sin ’2y the front lens n3 have been matched 2) Homogeneous for the centroid wavelength in such immersion, the smallest, still resolvable distance a way that the rays emitted by an ob- n2 = 1.518 (oil) n2 = 1.0 (air) between two object points, i.e. the ject point OP (Fig. 5) pass the immer- 2 Coverslip, ∆ n1 = 1.518 resolution limit 2 ymin, is sion film without being refracted and can thus be absorbed by the front OP Object(point) of refractive index nO ∆ ∆ nBR · sin ’ lens of the objective. In this case, the 2 ymin = 2 y’ Dry objective Immersion objective nOR · sin numerical aperture has been in- 1 2 5 creased by the factor n2, i. e. the 1.22 · b · · d wavelength and therefore the resolu- Fig. 4: 2∆y = min 2 · b · d · n · sin tion limit has decreased to 1/n . This The resolving power of the OR 2 microscope (according to means that the numerical aperture of R.W. Pohl, 1941) a dry objective and an immersion ob- 2∆y object, 2∆y’ image, (6) jective differs by the factor n , pro- 2 angular aperture 2 on the object side, vided the objectives can absorb rays 2’ angular aperture ∆ 0.61 · of the same angular aperture. The on the image side, 2 ymin = 2 resolvable angle size nOR · sin standard numerical apertures of im- on the object side, mersion objectives are 1.25 (water 2’ resolvable angle size The reciprocal value of (6) is de- immersion), 1.30 glycerin immersion) on the image side. scribed as resolving power, which and 1.40 (oil or homogeneous im- Fig. 5: should have as high a value as possi- mersion). The values correspond to Influence of the immersion ble. half the angular apertures = 56°, medium on the numerical 59° and 68°; for dry objectives, the aperture of the objective. 2 = Angular aperture The benefits numerical apertures would be re- of the objective of immersion duced to 0.83, 0.86 and 0.93. (numerical aperture = n2 • y sin ) 1 = Limit angle of The fundamental resolution formula Another advantage of immersion total reflection (6) valid for objects which are not objectives over dry objectives is their (= arcsin [n2/n1] self-luminous states that the resolu- considerable reduction or even entire 41°) reached; grazing light exit. tion limit depends on two factors, elimination of interfering reflected 2 = Total reflection namely the wavelength and the nu- light produced at the front surfaces merical aperture of the objective. If, of the coverslip and the front lens of therefore, the resolving power is to the objective. be increased or the resolution limit minimized accordingly, shorter wave- For the sake of completeness, we lengths and a larger numerical aper- would also like to mention the im- ture must be selected. mersion technique used to determine the refractive index of isolated solid bodies. The object to be measured is

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embedded in an immersion liquid, Useful (9a) the refractive index of which approxi- magnification -4 mates that of the object. Using a V · 5.5 · 10 [mm] -4 u = 5.8 · 10 heating and cooling stage, the tem- To enable the human eye to see two 500 · nAObj perature is then varied until the re- image points separately, an angular fractive index of the liquid is identical distance 2 of between 2 and 4 arc and to that of the object. The fact – de- minutes, or rived from the Dulong-Petit law – that the temperature dependence of (9b) the refractive index of solids is signifi- (7) -4 cantly lower than that of liquids is Vo · 5.5 · 10 [mm] -4 -4 ≤ ≤ -4 = 11.6 · 10 very important here. The refractive 5.8 · 10 2 11.6 · 10 500 · nAObj index can be determined using either the “Becke line” or, if high accuracies must exist between these points, or are required, by interferometric means; according to Ernst Abbe. If the lower please see the relevant literature. limit of the overall magnification of the microscope is Vu and the upper (9c) limit Vo, where the overall magnifica- ≈ tion of the microscope VM equals the Vu 500 nAObj quotient of the apparent visual range of 250 mm and the overall focal and length of the microscope fM, the cal- culation of Vu and Vo is easy: (9d)

≈ (8a) Vo 1000 nAObj

2∆y 2∆y [mm] V -4 Therefore, the performance of the = u = 5.8 · 10 (= 2’) f 250 microscope is meaningfully utilized 2) Due to the matching M refractive indices of the only if the selected total magnifica- front lens, the immer- tion is no less than 500x and no sion liquid and the (8b) more than 1000x the numerical aper- cover slip, the micro- scopist first tends not ture of the objective to make any difference 2∆y 2∆y [mm] V -4 Our forefathers rightfully termed = o = 11.6 · 10 (= 4’) between coverslip- f 250 magnifications > 1000 nA as corrected (e. g. HI M Obj 100x/1,40 ∞/0.17) “empty magnifications” because still and non-corrected with = 550 nm = 5.5 · 10-4 mm smaller object details can no longer ∞ (e. g. HI 100x/1.40 /0) be expected to be resolved, which immersion objectives. This is certainly possible and will result in ineffective over-magnifi- in monochromatic light cation. ∆ (centroid wavelength); 2 ymin = = in polychromatic light, 2 sin 2 nAObj however, immersion Rainer Danz, Carl Zeiss AG, liquid and coverslip finally result in Göttingen Plant usually display different [email protected] dispersions, i. e. the refractive indices depend on the wavelength to a greater or less extent. This effect becomes apparent in the microscope image with chromatic and spherical aberration. Therefore: always carefully note the correction state of the objective!

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Numerical details Aperture Resolving The numerical aperture indicates the objective). The formula for the nu- power resolving power of an objective. Put merical aperture shows that, in addi- more simply, the resolving power of tion to the angular aperture of the The resolving power of an an objective depends on how much objective, the refractive index of the objective is the ability to light of a specimen structure reaches medium between the coverslip and show two object details the objective. This amount of light the objective is also used for the separately from each other depends on what is called the angu- computation. in the microscope image. lar aperture of the objective. The The numerical aperture of the larger the angular aperture, the bet- In the case of dry objectives, the objective directly determines ter an objective will resolve the delight rays are refracted away from the the resolving power: the tails of a specimen. However, it is not perpendicular on entering the air be- higher the numerical aper- the angular aperture which is speci- tween the coverslip and the objective ture, the better the resolving fied on the objective, but the numeri- in accordance with the refraction power. cal aperture. law. Therefore, strongly inclined light rays no longer reach the objective The theoretically possible The numerical aperture is defined and do not contribute to the resolu- resolution in light microscopy by the formula A = n * sin (A: nu- tion. With oil immersion objectives, is approx. 0.20 µm. The merical aperture, n: refractive index immersion oil is inserted between the resolving power of an objec- of the medium between the coverslip coverslip and the objective: even tive is defined by the formula and the front lens of the objective, strongly inclined light rays reach the sin : half the aperture angle of the objective. d = 2 x A

d: distance between two Front lens of the objective Front lens of the objective image points : wavelength of light A: numerical aperture of the objective

Air Immersion oil Refractive index Cover slip of various media Dry objective Oil immersion objective

Refractive index n of the media through which the light ray passes in the beam path of a microscope.

Air n = 1.000 Objective with small Objective Glass n = 1.513 angular with large Oil n = 1.516 aperture angular aperture

Object plane

2 2

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Highlights from the History of Immersion

Robert Hooke (1635-1703) was the this problem during Brewster’s time. first to discuss the technique of Because microscope slides were very immersion: “that if you would have a expensive at the time, microscopists microscope with one single refrac- in the 19th century did not yet accept tion, and consequently capable of oil immersion. Amici gave up on oil the greatest clearness and brightness, immersion and converted to water spread a little of the fluid to be ex- immersion. A short time later in details amined on a glass plate, bring this 1853, he built the first water immer- under one of the globules, and then sion objective and presented it in Immersion oil move it gently upward till the fluid Paris in 1855. touches and adheres to the globule.” In 1858, Robert Tolles (1822- In the beginning, natural cedar His “Lectures and Collections” from 1883) built his first water immersion oil was used. Gradual thicken- 1678, published in his book “Micro- objective which had two exchange- ing leads to alteration of the scopium” in the same year, thus able front lenses: one for working refractive index over time. marked the beginning of oil immer- under dry conditions, the other for Exposed to air, it turns to resin sion objectives. water immersion. and becomes solid. Sir David Brewster (1781-1868) Approximately 15 years later in Nowadays, synthetic immersion proposed the immersion of the ob- 1873, he constructed his famous oil with a constant refractive jective in 1812. Around 1840, Gio- 1/10 objective for homogenous im- index is used. It does not harden vanni Battista Amici (1786-1868) pro- mersion. Edmund Hartnack (1826- in air and can therefore be duced the first immersion objectives 1891), who in 1859 presented his stored longer. that were used with anis oil and had first water immersion objective, also the same refractive index as glass. added a correction ring for the first However, this type of immersion was time. Hartnack sold around 400 not yet used to increase the aperture, models over the next five years. By but more to correct chromatic aber- 1860, many German microscope ration. Amici had already recognized manufacturers offered water immer-

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details

Glycerin

1,2,3 propanetriol – is the most simple tertiary alcohol. The Greek word glykerós means “sweet”. The viscous, hygroscopic, sweet- tasting liquid boils at 290 °C and freezes at 18 °C. Glycerin can be mixed with water and lower-order alcohols. A mixture of water and glycerin is used in microscopy for immersion. It is mainly used in UV microscopy as glycerin transmits UV light.

Objectives

sion lenses, including Bruno Hasert in were offered in the Zeiss catalog at “On New Methods for Improving Eisenach, Kellner in Wetzlar, G&S the time, all with an aperture of Spherical Correction” in the “Royal Merz in Munich and Hugo Schroder 180°. They had different working dis- Microscopical Society” magazine. In in Hamburg. Hartnack’s immersion tances, but also a numerical aperture this article, Abbe described the optics lenses, however, were considered the of 1.0. Objective no. 3 was equipped he used in his 1873 experiments. He best. with a correction ring. also added that homogenous immer- At the “Exposition Universelle” in In August 1873, Robert Tolles built sion systems make it possible to 1867 in Paris, Ernst Grundlach (1834- a three-lens objective for homoge- achieve an aperture at the limits of 1908) presented his new glycerin im- nous immersion in balsam with a nu- the optical materials used and avail- mersion lens, claiming that he devel- merical aperture of 1.25. It was the able at the time. Robert Koch was oped the lens because he wanted to first homogenous immersion system one of the first to utilize the Abbe oil use an immersion medium with a for microscopes to be recognized at immersion objective and the Abbe higher refractive index than water. the time. In the same month, he pro- condenser system for research pur- In 1871, Tolles once again pre- duced his first objective for glycerin poses. In 1904, Carl Zeiss manu- sented something new: he used immersion with a numerical aperture factured its 10,000th objective for Canada balsam as an immersion of 1.27. homogenous oil immersion. medium for homogenous immersion. In August 1877, Carl Zeiss began His discovery that Canada balsam has building Abbe’s oil immersion objec- the same refractive index as the tives which later became known as crown glass that was standard at “homogenous” immersion. The de- the time remained unused until Ernst sign of the Zeiss oil immersion objec- Abbe discovered a suitable liquid in tives was influenced by the work of 1877. The Zeiss Optical Works in Je- J. W. Stephenson, which Abbe em- na also produced initial water immer- phasized in a lecture to the Jena Soci- sion objectives in 1871. In 1872, Carl ety for Medicine and Natural Science Zeiss introduced the Abbe water im- in 1879. mersion objective. Three objectives In 1879, Ernst Abbe published his

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From the History of Microscopy: Abbe’s

The history of microscopy started Until 1866 – the year when the co- Abbe’s key theory around 1590 with Dutch spectacle operation between Carl Zeiss and on microscope image makers. The first “simple” micro- Ernst Abbe began – microscopes, and formation scopes date back to Antony von the microscope objectives in particu- Leeuwenhoek (1632-1723) and lar, were made by trial and error, In the end, it was Carl Zeiss who rec- his contemporaries. Leeuwenhoek resulting not only in some micro- ognized the importance of a solid built microscopes with a single, scopes with outstanding optical per- theoretical basis and who initiated small lens displaying magnifica- formance, but also in some with less and also financed Abbe’s research. tions of up to 270x. This enabled desirable features. Carl Zeiss (1816- The major breakthrough for the him to discover protozoa (single- 1905) and Ernst Abbe were aware building of microscopes came with celled organisms) as early as that optimum and – above all – the theory of microscope image 1683. The “combined” micro- consistent performance would only formation from Ernst Abbe (1840- scopes are attributed to the Eng- be possible on a sound theoretical 1905). After countless calculations lishman Robert Hooke (1635- basis. The first calculations were and experiments, Abbe realized that 1703), who is known to have made of the geometric beam path. it is the diffraction image in the used these for the first time. To improve correction, Abbe used back focal plane of the objective “Combined” microscopes consist lower apertures than those of the that is decisive for image formation. of an objective lens and an eye- objectives made by Zeiss until then. In 1873, he wrote: “No microscope piece. Hooke already recognized The results were not really satisfacto- permits components (or the features the importance of microscope illu- ry: fine specimen structures remained of an existing structure) to be seen mination at that time. However, blurred, and their resolution was less separately if these are so close to the aberrations of both micro- good than that obtained with the each other that even the first light scope types impaired precise old objectives with a wider angular bundle created by diffraction can observation. Nevertheless, they aperture. no longer enter the objective simul- formed the base of the pioneer- taneously with the non-diffracted ing microscopic discoveries in the light cone.” 19th and 20th centuries.

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Diffraction Trials

This is also the core of Abbe’s theory of microscope image formation. His theory, based on the wave charac- teristics of light, shows that the maximum resolution is determined by half the wavelength of the light used, divided by half the numerical aperture. Abbe’s theory of image formation and resolution limits was rejected by many biologists and mainly by microscopists in England, who were too Microscope stand VII. Microscope stand I. biased by the old microscopy technique of using strongly magnifying eyepieces, a method known as empty magnification. Finally, Abbe was able to prove his theory with a system of experiments he first demonstrated using the ZEISS Microscope VII. Robert Koch used the same Microscope VII for his discovery of the tuberculosis bacterium.

Cover page of brochure on Abbe’s illumination apparatus. the diffraction apparatus.

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1 3

2 4

Diffraction experiments

Even today, diffraction experiments still play a major role in training courses in microscopy, and make a major contribution to the understanding of modern microscopy 6 techniques. Abbe created almost 60 experiments to prove his theory of image formation. A diffraction plate with various objects engraved in the form of lines and dots is used as a specimen. The condenser is removed so that the light source lies at infinity and can be made to adopt a point- shaped structure by closing the luminous-field diaphragm (Fig. 1). Re- 5 7 moval of the eyepiece or the use of an auxiliary microscope permits viewing of the images produced in the back focal plane of the objective.

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Fig. 10: Abbe’s diffraction apparatus b.

9 10

Single slit Line grid with is twice as distant from the 0th 16 µm grating maximum as in the 16 µm grid. With a single slit (Fig. 2), a light strip constant (Fig. 3) appears in the back focal Point grid plane of the objective, which is per- In the produced diffraction image of pendicular to the optical axis and the 16 µm line grid (Fig. 5), the im- In the diffraction image of the point displays the point-shaped light source age of the light source, also called grid (Fig. 8), the respective primary in the center. This light strip is pro- the zeroth maximum, and the first diffraction spectrum (Fig. 9) can be duced by the diffracted light waves and second order secondary maxima seen in the back focal plane of the at the edges of the slit. are clearly separated from each other objective. From this, Abbe concluded and imaged much more sharply (Fig. that each specimen forms a specific Double slit 6) than is the case in Fig. 4. Further- diffraction image of the light source. more, it is also evident that blue light The double slit (Fig. 2) enables obser- is less diffracted than red light. Influencing the vation of the interference phenome- diffraction image non: the image of the light source is Line grid with located in the center, while bright 8 µm grating If an intermediate component with a and dark sections extend to the right constant slit is inserted between the objective and left in a regular sequence (Fig. and the nosepiece (Fig. 10), in which 4). The bright sections display charac- In the diffraction image of the 16 µm various stops can be inserted, parts teristic color fringes (spectral colors). line grid (Fig. 5), the 1st secondary of the diffraction image can be Abbe designated this image in the maximum is twice as distant from the faded out. For more precise orienta- back focal plane of the objective as 0th maximum, and the 2nd secondary tion, the intermediate component the diffraction spectrum. maximum is no longer visible at all can be rotated through 360 degrees. (Fig. 7). From this, Abbe concluded This means that the primary diffrac- that the closer the structures – or tion image is changed by artificial lines in this case – the more the light means. waves are diffracted, explaining why the 1st maximum in the 8 µm grid

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14 11 12

Point grid In a further experiment, Abbe then concentrate on the correction of demonstrated why the resolution spherical and chromatic aberration. From the diffraction image of the formula is He realized that this requires the point grid (Fig. 9), the 0th and the development of special glass materi- first secondary maxima in the hori- als. His collaboration with the glass d= 2 x n x sin = zontal axis are faded out via the slit. 2nx sin maker Otto Schott (1851-1935) start- In the intermediate image, a line grid ed in 1879, and the first Apochromat then appears in the perpendicular numerical aperture. objectives featuring high color fidelity direction (Fig. 11), and at an angle of were already launched in 1886. 45° when the slit is oriented diago- Abbe pushed the point-shaped light In 1893, August Köhler (1866- nally (Fig. 12). A particularly remark- source to the edge (known as 1948) developed his illumination able feature of the artificially pro- oblique illumination): therefore, the technique with separate control of duced 45° grating is that the lines are 1st secondary maximum can be twice luminous-field diaphragm and con- closer to each other than in other im- as distant as in straight illumination, denser aperture on the basis of ages. This is bound to be the case be- i. e. the distance d between two Abbe’s results. Köhler also developed cause the secondary maxima in the points or lines can be twice as small. the microscope with ultraviolet light, 45° diffraction image are further On this basis, Abbe had already de- which was primarily built to increase away from the 0th maximum than in veloped the illumination apparatus the resolution by a factor of 2 relative the perpendicular or horizontal dif- with focusing condenser in 1872 (Fig. to green light. Finally, the phase fraction images. 14). In today’s microscopy, we use uni- contrast technique from the Dutch With these experiments, Abbe fi- lateral oblique illumination to achieve physicist Frits Zernike also is attribut- nally showed that the image in the full resolution with the maximum able to manipulation in the back microscope is created in the space condenser aperture. focal plane of the objective. between the primary diffraction im- The theory of microscope image With his theory and experiments, age (back focal plane of the objec- formation and its practical implemen- Ernst Abbe decisively shaped the de- tive) and the intermediate image tation defined the resolution limits velopment of microscopy in the 19th plane through interference of dif- and thus enabled the scientific con- and 20th centuries. Numerous Nobel fracted light waves. struction of microscopes. Abbe could prizes are directly or indirectly con-

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nected with microscopy. Abbe himself was twice nominated for the Nobel prize. Pioneering examinations in cell and molecular biology would not have been possible without modern light microscopy. Metallogra- phy would probably not have achieved its current status without microscopy, and semiconductor technology would probably not even exist at all.

Heinz Gundlach, Heidenheim

Image plane

Image of the specimen

Back focal plane

Diffraction image of the specimen Specimen plane

Specimen 13

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The Science of Light

The importance of light is empha- in the 6th century BC: here the em- the object has on the eye by means sized in the First Book of Moses in phasis was on explaining the effect of the medium between them (“the the Bible: “In the beginning, God the visible object had on the eye. The transparent”). created the heaven and the earth. various schools developed ideas that In addition to the process of see- And the earth was without form, differed from each other to a greater ing in itself, the Greeks also studied and void; and darkness was upon or lesser extent and were generally the laws of geometric optics. It seems the face of the deep. And the rather imprecise. that Plato (424-347 BC) was aware Spirit of God moved upon the The predominant theory in An- of the law of reflection and he de- face of the waters. And God said, cient Greece was the extramission scribed the reflection of concave and Let there be light: and there was theory, which can presumably be cylindrical mirrors. The mention of light. And God saw the light, that traced back to Pythagoras (570/560- oars bending in water indicates that it was good: and God divided the 480 BC) and was later supported the phenomenon of refraction was light from the darkness.” Without in particular by Euclid (around 300 also familiar. The playwright Aristo- light life is impossible. Light plays BC) and Ptolemaeus (around 100- phanes (445-385 BC) described the an important role in our lives. 160 AD). The extramission theory effect of burning glasses (glass lenses And the question of the “nature assumed that we are able to see as or glass globes filled with water). of light” is one that we have a result of hot rays that emanate Ptolemaeus, who summarized the always endeavored to answer. from our eyes towards an object. The entire optical knowledge of the an- resistance these rays meet with when cient world and systematically exam- they reach the cold object causes ined the refraction of light, is perhaps them to be sent back, enabling the the most important optical scientist Besides mechanics, optics would information they have gathered to of that time. seem to be the oldest field in which reach the eye. The ability of many an- During the Middle Ages, Chris- scientific work has been conducted. imals to see at night was put forward tianity was not particularly open to The Babylonians, on the basis of their in support of this theory. science. It was the Arabs who not experience, were already applying Aristotle (384-322 BC), in particu- only collected and translated the an- the law of the rectilinear propagation lar, had a different view. He believed cient writings but also made their of light around 5000 BC in the use of that light is not something physical own scientific contributions. The astronomical instruments. that moves between the object and most important Arab scientist was There is evidence of scientific the eye, but rather that the process Abu Ali Al-Hasan Ibn Al-Haitham study in the field of optics in Greece of seeing is the result of the effect (965-1040), also known as Alhazen,

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who wrote more than 200 works on regarded as the founder of modern optics, astronomy and mathematics. optics. He succeeded in correctly ex- In Huygens’ famous principle, His principal work, the “Book of Op- plaining how the camera obscura every point on a forward- tics”, contained descriptions and ex- and the eye worked, including the moving wavefront is seen as planations regarding light and vision. lens and the retina. Thomas Harriott a source of new waves. He used From the 12th century, the main (1560-1621) was supposedly the first this principle to develop the scientific focus shifted geographically person to discover the law of refrac- wave theory of light. With from the East back to the West of tion. a new method (1655) for the the known world. Initially, however, In 1637, Rene Descartes (1596- grinding and polishing of lens- only the works of Alhazen, Ptole- 1650) derived a theory relating to the es, Huygens achieved an im- maeus and Euclid were translated Law of Refraction in his work “La provement in optical perform- and summarized. There is evidence Dioptrique”. He was one of the first ance. This enabled him to dis- Christian Huygens that Roger Bacon (1214-1294), a Do- to attempt to explain all optical laws cover the moon of Saturn and (1629-1695) minican monk, studied the camera and phenomena on the basis of the to make the first exact descrip- Dutch astronomer, obscura, which he recommended for mechanical properties of the light tion of the rings of Saturn. mathematician, physicist and clockmaker. observing solar eclipses. Bacon also source and the transparent medium. To observe the night sky, Huy- predicted the development of eye- The work of Johannes Marcus Marci gens developed the pendulum glasses and the telescope. Eyeglasses de Kronland (1595-1667) and clock with an exact time meas- were apparently invented in Italy at Francesco Maria Grimaldi (1618- ure. In 1656 he invented the the end of the 13th century. 1663) gradually brought us closer to Huygens telescope. He devel- At first, however, it was not the wave theory of light, which was oped theories about centrifugal known how they functioned exactly, then supported emphatically by force in circular motion. These Birefringent calcite. since people did not know how the Robert Hooke (1635-1703). At the helped the English physicist eye saw, nor how lenses worked. same time as Hooke, the Jesuit priest Sir Isaac Newton to formulate Giovanni Battista della Porta (1535- Ignace Gaston Pardies championed the law of gravitation. In 1615) compared the eye with a cam- the theory of the wave nature of 1678 Huygens discovered the era obscura. Father Franciscus Mau- light. Nevertheless, it is Christiaan polarization of light through rolycus (1494-1575) recognized that Huygens (1629-1695), with the Huy- birefringence in calcite. defective vision was caused by the gens’ principle that was named after incorrect curvature of the lens. him, who is regarded as the true Johannes Kepler (1571-1630) is founder of the wave theory of light.

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Stazione Zoologica Anton Dohrn, Naples,

Fig. 1: Group of researchers at the Stazione Zoologica at 1 the end of the 19th century.

Since its inception, the zoological Naturalists as well as philosophers wealth of life forms, particularly sim- station has been devoted to basic have always shared a fascination for ple forms, caused many scientists to research in biology for all its in- life under the sea. This curiosity, sci- see the oceans as a source of biologi- ter-disciplinary activities – from entific observation and legends cal models, experimental objects and evolution and molecular biology passed down through the years, led as a metaphor for fundamental bio- to ecophysiology. The station cel- Pliny the Elder (23-79 A. D.) to pen logical problems such as the organi- ebrated its 125th anniversary in Naturalis Historia. It attracted atten- zational plan of life, embryogenesis, 1998. Named after founder, finan- tion to sea life, an unending source general physiology, evolution and cier and first director, Anton of wondrous (Mirabilia) and fantastic phylogeny. Dohrn, the center took 18 months monsters, a place of mystery and Johannes Müller (1801-1858), con- to complete, finishing in Septem- a constant source of life and beauty. sidered the founder of physiology ber 1873. Located in Naples, Italy, To the German science scene in the and theoretical biology, raised inter- the station served as an example 19th century, which was marked by est in marine organisms for the un- for many other marine biological men such as Alexander von Hum- derstanding of fundamental biologi- centers and institutes. This boldt, the ocean was a place of ele- cal problems. The concept of marine renowned list includes Woods mentary life forms and a symbol of biology research published by Müller Hole in the USA and Misaki in the endless search for knowledge. was a means of explaining the funda- Japan, as well as the Kaiser Wil- Marine life became the focus of mental biological concept. The sta- helm Institutes which later be- interest of all those who recognized tion’s first boat carried his name as came the Max Planck Institutes. A the philosophy of nature. Appearing an indication of its purpose. It was novelty when the station was in the “protoplasmic theory of life”, used to gather marine plants and founded, its internationality was initial forms of cell theory looked for animals. fostered and ensured by influen- “elementary forms of matter”, the tial scientists at the time, Charles “primordial mud” teeming with one- First marine Darwin and Rudolf Virchow, celled organisms, in the depths of the institutes among others. oceans. The new generation of biologists from the 1850s and 1860s was Zoologist and parasitologist P. M. van well versed in “natural philosophy” Beneden from Leuven, Netherlands and in Darwinism. They viewed the founded the first marine laboratory in ocean as a source of knowledge and Ostende 1843. Similar initiatives in as an experiment of life. In the sec- North America emphasized the need ond half of the 19th century, the for such research institutes: Louis

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Italy

Agassiz founded the Anderson 1875 and Anton Dohrn signed the The station’s School of Natural History in 1873; contract with the City of Naples. technical setup Johns Hopkins University the Chesa- peake Zoological Laboratory in 1878. Anton Dohrn’s The station has always provided sci- The Woods Hole Oceanographic In- idea of independence entists with state-of-the-art research stitution opened in 1892. equipment which was acquired either However, these were all university In order to increase the international- as a gift or at a very low price. The and institute field stations. They can ity of the center, and to ensure the latest developments from Carl Zeiss also be split up into two categories: economic – and therefore political – were used, tested and presented to centers such as Naples were dedicat- independence and freedom of re- the guest scientists. Ernst Abbe, one ed to research and practical training, search, Anton Dohrn introduced a of Anton Dohrn’s few close friends while French and American institutes series of innovative measures to made it possible for the station to were used primarily for teaching. finance projects. The system for purchase Zeiss microscopes and other renting work and research space optical instruments at cut-rate prices. Deciding on Naples deserves to be mentioned first. For Station employees not only assisted an annual fee, partners such as uni- in the optimization of the instru- During his stay at the zoological cen- versities, governments, private insti- ments, but also aided in distribution ter in Messina, Anton Dohrn realized tutions, and even individuals were to the international science commu- that a permanent laboratory struc- permitted to send a scientist to nity. Employees and guest scientists ture was required to study ocean life. Naples for one year. Everything need- constantly improved the scientific This is where his dream of a “full ed for unlimited research was avail- methods, techniques and instruments house” for ocean research started: able. Scientists could even use the provided to the station. instant and permanent availability of knowledge of station employees. instruments, laboratory stations, lab- Everyone was free to pursue their oratory services, chemicals, books projects and ideas. The fast and free Gaius Plinius Secundus, better and much more. exchange of ideas, methods, tech- known as Pliny the Elder The diversity of life in the Gulf of niques and instruments, as well as (Latin: Plinius maior), an Naples, the size of the city and its in- the contact between scientists from ancient author and scientist, ternational reputation contributed to different cultures, was decisive for is best known for his scientific Dohrn’s decision to locate his center the success of the system. In 1890, work Naturalis Historia. He in Naples. With a mixture of imagina- for example, 15 countries rented 36 was born around 23 A. D. in tion, willpower, diplomatic skill, luck tables for one year. More than 2,200 Novum Comum and died on and some friendly help from other scientists from Europe and America August 24, 79, in Stabiae. The scientists, artists and musicians, he had worked at the center by the time Naturalis Historia (occasionally was able to remove any doubts, un- of Dohrn’s death in 1909. Historia Naturalis) comprises certainties and misunderstandings, Internationality increased as a re- a comprehensive encyclopedia and convince the city to donate a sult of scientific publications. These of natural sciences and re- piece of land directly beside the sea: included Notes from the Zoological search. It is the oldest known a piece of land at one of the most Station in Naples (1879-1915), which complete systematic encyclo- beautiful places in Naples – Park Ville later became the Pubblicazioni della pedia. The Naturalis Historia Reale. He promised to finance Stazione Zoologica di Napoli (1924- consists of 37 books with a the zoological center himself. Dohrn 1978) and has appeared as Marine total of 2,493 chapters. The knew exactly what he wanted. The Ecology since 1980. The Zoological bibliography cites almost 500 cornerstone was laid in March 1872 Annual Report (1880-1915) also ap- authors, including 100 primary and construction of the station was peared for a short time. The Fauna and almost 400 secondary completed in September 1873. He and Flora of the Gulf of Naples sources. and his father paid for two-thirds of monograph is still considered an the construction costs. The remainder outstanding work today. was financed by loans from friends. Although the center had just opened its doors, the first scientists headed to Naples in September 1873. The of- ficial inauguration was on April 14,

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Well-known visitors debated by physiologists and histolo- The Stazione gists at the time, the relationship be- Zoologica organism The zoological station has never had tween ganglion cells and nerve its own research project. The struc- fibers, the nature of nerve impulses To this day, many complementary ture of the center always reflected and brain functions at a cellular level facts contribute to the uniqueness of the interests of its guests. According later became a traditional field of in- the station: the high level of scientific to statements by Theodor Boveri, terest. A visit by Robert Koch to activity, the active and constant ex- Dohrn always had an eye open for Naples in 1887 convinced Dohrn of change of information with the inter- the importance of the different scien- the necessity of establishing a bacte- national scientific community, the tific aspects and their interaction in riological laboratory. Developmental flexible organizational structure and the overall scheme of things. This biology examinations on sea urchins the associated independence of was the only way to attract the best were first conducted by Theodor political and academic institutions, scientists of the time to Naples. Boveri in 1889 and later by Otto the incomparable library, the avail- Nobel Prize winner Fridtjof Nansen Warburg. ability of state-of-the-art instruments, was accepted by Dohrn in 1886 al- the cultural atmosphere and the though he did not have a financial Art and culture in creative dialogue between different agreement with Norway. Nansen a scientific facility cultures. Tradition and innovation “found” a new field of interest: hotly have merged right from the begin- The significance of the zoological ning of the station and enabled the station extends far beyond pure sci- Stazione Zoologica “organism” to Fig. 2: entific aspects. It is also known for survive until now. The zoological Delivery note/invoice its humanitarian values and its cul- station has been officially called for Microscope Stand IVa No. 29057 from April 20, tural climate. For the majority of the “Stazione Zoologica Anton Dohrn” 1898. many guest scientists, the “Naples since 1982 and has almost 300 experiment” was an impressive mix- employees. Fig. 3: Theodor Boveri (1862-1915): ture of new research, human experi- the development of doubly ence, acquiring new methods and (di-sperm) fertilized sea the exchange of ideas and cultural urchin eggs. differences. The zoological station is the only institution in the world where science, art and music were integral elements of a unique project, the complementary halves of a 2 dream, from the beginning. The ar- chitectural design and the technical- scientific equipment are a perfect match. Art and music were an essential element of cultural life in the 19th century. Dohrn wrote to E. B Wilson in 1900: “Phylogeny is a sub- tle thing. It requires not only the an- alytical powers of the researcher, but also the constructive imagination of the artist. Both must balance each other out, otherwise it does not suc- ceed.”

www.szn.it 3

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Felix Anton Dohrn

Anton Dohrn founded the zoo- Dohrn studied in Königsberg, Bonn logical station in Naples in 1870. and Jena under Rudolf Virchow, Ernst For decades it was the leading Haeckel and Carl Gegenbaur. Follo- international center for marine wing his studies in medicine and zoo- research. logy, he became interested in the theories of Darwin. In 1870, he foun- Anton Dohrn was a zoologist and ded the zoological station for the one of the first outstanding resear- research of marine fauna in Naples, chers of phylogeny. He came from a Italy, one of the first marine research well-to-do family in the German city centers. He also studied the phyloge- Felix Anton Dohrn of Stettin. In his early childhood, he ny of arthropods based on embryolo- learned that art and science coexist gical and comparable anatomical da- Born December 29, 1840, and interact: his father Carl August ta. Building on his insights, he was in Stettin, Died September 26, 1909, (1806-1892) corresponded with ar- the first to suggest that vertebrates in Munich tists, poets and scientists such as evolved from annelids. Furthermore, Alexander von Humboldt and Felix he described the “principle of the Mendelsohn. His children were requi- change of functions”. “…not only that Abbe provid- red to “know their Goethe”, be Dohrn earned his doctorate in ed new instruments at knowledgeable about music and 1865 in Breslau. He qualified as a a very low price, or even share his passion for science. lecturer in 1868 in Jena and taught donated them, he also re- zoology until 1870. From 1874 until ceived suggestions from there, his death, he was director of the ma- the control of the practice, rine research station in Naples, Italy the critical thoughts of experi- (Stazione Zoologica di Napoli). In ence. The station was also 1874 he married 16 year old Marie a place where foreign re- de Baranowska. searchers were introduced to Despite his nationality and cultural and acquired Zeiss equipment, origins, Anton Dohrn received con- and then announced the siderable support from the British splendor of superior German natural science tradition during the workmanship in their home realization of his dream of founding countries. The station was a marine biology laboratory. At the almost a type of ideal export annual meeting of the British society warehouse and Dohrn was in 1870 in Liverpool, a committee delighted to write to Abbe was founded with the intention of that Balfour acquired this Anton Dohrn, 1898, promoting the foundation of zoologi- apparatus and he wants one drawing by cal stations in different regions of the for everyone…” Johannes Martini. world. In fact, it was this committee that increased the fame of Dohrn’s Theodor Heuss was friends zoological station in Naples in the En- with Dohrn’s son Boguslav glish-speaking world with regularly who, on the 100th birthday of published reports and articles in Na- the great natural scientist in ture. 1940, requested that Heuss write a comprehensive summary of the life work of Anton Dohrn.

From “Anton Dohrn: A Life for Science” by Theodor Heuss (1940)

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Fig. 1: View of the hall of frescoes at the Stazione Zoologica Anton Dohrn.

Fig. 2: Desk in front of the east wall of the hall of frescoes with the “La Pergola” fresco.

1 The Hall of Frescoes

An unexpected treasure is hidden Two German artists, Hans von his meeting with friends of Anton above the aquarium and between Marées, and sculptor and architect Dohrn, with the world of the an- the labs, technical equipment and Adolf von Hildebrand, decorated it tiques and the sunny life on the offices at the Zoological Station with a fresco cycle that plays a Mediterranean. in Naples: the Hall of Frescos, a unique role in the history of 19th Hilderbrand studied in Nuremburg room Anton Dohrn dedicated to century art. and Munich. In 1867 he accompa- music and entertainment. Hans von Marées (1837-1887) nied his teacher, von Zumbusch, to was one of the most influential Ger- Rome. From 1872 to 1897, he lived man artists in the second half of the in Florence and focused his work on 19th century. He developed an ideal- the sculpture of the Italian Renais- istic style of painting with clarity of sance. With his marked tectonic form focused on people. As a painter talent, he created fountains and he was attracted to the world of the monuments. antiques. As an art theoretician, he worked together with Adolf von Hildebrand (1847-1921), the leading sculptor of his time, on the theory of pure visibility. Following his studies and training in Berlin (1853-1855) and Munich (from 1857), and travels to Italy, Spain and France (1864-1866), Marées completed the frescoes in Naples which represent a high point Christiane Groeben, [email protected] www.szn.it 2 of his work. They are the result of

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Bella Napoli

During his trip to Italy in 1786, urbs around three million. It is situat- details Johann Wolfgang von Goethe ed halfway between Mount Vesuvius also made a stop in Naples: and another volcanic region, the See Naples “Everyone is on the street, sitting Campi Flegrei (Phlegraean Fields) on and die in the sun, as long as it shines. the Gulf of Naples. The Neapolitans believe they are Science quickly found a home in “See Naples and die,” is an living in paradise,” he wrote in the city: in 1224, Friedrich II von Ho- expression often used when February 1787. henstaufen founded the University of someone is completely mesmer- Naples. For some, it is the loudest, ized by the beauty of something Naples is well known, beloved and most polluted and chaotic city; for they have seen. popular and attracts many visitors. It others it is the most beautiful and It is based on the Italian saying 1 is a city with its own character, a city lively. Five and six story apartment “Vedi Napoli e poi muori”. of enchantment, entrapping every- buildings existed as early as the 16th In Italian it has a funny double Fig. 1: one in its spell with the beauty of the century. It was the largest city in Eu- meaning. It is a play on words Menu from 1907 with the sea, the magic of history, the extra- rope and space was at a premium. with the name of a city, Stazione Zoologica. ordinary architecture and its friendly Scholars came to the city at all times “Muori”, that is located just people. to revel in its artistic splendor. beyond Naples which can only Fig. 2: The Stazione Zoologica The city was founded sometime in Naples is considered the birthplace be seen upon leaving, and the around 1873. the 8th century B. C., most likely by of pizza. The recipe for a margherita verb form “muori”, meaning to inhabitants of the Greek Cumae has remained unchanged since the die. Enjoying a favorable climate, colony. In the 17th century, Naples 16th century: topped with only Naples is a special place: the boasted 300,000 citizens, making it tomato sauce, mozzarella and basil, Italians consider it a piece of the second largest European city the pizzaiolo places the culinary heaven on earth, while the after London. Today, Naples (Greek: delight into the wood-burning brick Germans and French saw it as nea polis: New City, Italian: Napoli) is oven. Three minutes later, it is ready the center of sorcery and black the third largest city in Italy after to eat and the next one takes its magic until the 19th century. Rome and Milan, and is the largest place. city in southern Italy. It is the capital of the Campania region. The city currently has approximately one million inhabitants; together with the sub-

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From Users

The Zebra Fish as a Model Organism for

Fig. 1: The zebra fish (danio rerio) is 3-day-old zebra fish very easy to breed, requiring only in red and green fluorescence: antibody three days to develop from an labeled axon populations egg into a free-swimming larva. and GFP motor neurons As the zebra fish remains trans- NeoLumar S 1.5x 150x magnification. parent throughout its entire developmental period, it is an ideal organism for examinations of ver- Specimen: tebrate organ development under Prof. M. Bastmeyer, the microscope. Examinations on Dr.M.Marx,Friedrich Schiller University zebra fish models lead to a better Jena, Germany. understanding of organ development of “man the vertebrate” Photo: Dr. M. Zölffel, Carl Zeiss. and his diseases.

In cancer research For retinal diseases The First Stereo- microscope The zebra fish has already replaced Degenerative changes to the retina Came from Jena the well-established model organism are genetic diseases in people that – the mouse – in cancer research: the cause photosensitive receptor cells It all started at the Weimar Courtyard mouse has a longer developmental to die. This is one of the most com- in Jena in 1892. Under the leadership cycle and the ontogenesis stages mon causes of blindness in humans. of Ernst Abbe and developmental are less translucent than in the Zebra fish suffer from similar genetic biologist Ernst Haeckel, this was zebra fish. Until now, mice were used eye diseases. The development of the regular meeting point for that develop cancer cells (e. g. the fish eye and the activation of the science employees from the universi- leukemia) caused by genetic muta- nerve fibers in the zebra fish’s eye ty and the Zeiss Works. At one of tions. These cells are transfected are very similar to the human eye. these gatherings, American zoologist with GFP using molecular biology The short development period of Horatio S. Greenough expressed his techniques. The extremely powerful the zebra fish permits observation of wish for a ”binocular microscope SteREO Lumar.V12 fluorescence these degenerative retinal processes that renders true 3D images.” stereo microscope enables scientists with the SteREO Discovery.V12 and Carl Zeiss set to work on fulfilling to optimally view and research the Lumar.V12 high-resolution stereo- this wish and constructed the first progression of the disease. microscopes as if in slow motion, industrially manufactured stereo- thus enabling better research into microscope at the end of 1897 – the the cause of blindness and possible Greenough double microscope. treatment methods. The eyesight of zebra fish larva is examined using a special test. The stereomicroscope makes it possible to examine the developing eyes of blind as well as normally sighted fish and also compare them to each other.

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r Developmental Biology

2 3

Fig. 2: Sketch of Greenough’s idea for a binocular microscope that renders true 3D images.

Fig. 3: The Greenough double microscope from Carl Zeiss. 4 Fig. 4: It had two tubes tilted towards each marine biology: the Greenough stere- Dissecting microscope other at a convergence angle of 14 omicroscope enabled exact research following the design of Paul Mayer. degrees with objective lenses at the into the lifecycle of many inverte- lower ends. Carl Zeiss ensured that brates (e. g. polyps, bristle worms, Fig. 5: the axes on the two lenses were in snails) for the first time. It also con- SteREO Lumar.V12. one plane, i.e. they actually intersect- tributed considerably to the most im- ed. Porro erecting prisms were used portant discoveries in developmental between the lenses and the eye- biology and genetics of the early 20th pieces. These prisms ensure that im- century (Wilhelm Roux, Hans Spe- ages are upright and unreversed, i. e. mann, Thomas Hunt Morgan). the images can be viewed as they Today, the SteREO Lumar. V12 is are in reality. This was also a demand setting new standards for the fluores- from Greenough and the guarantee cence microscope examination of of a true orthoscopic impression complex issues related to develop- when looking through the stereo- mental genetics in biological and microscope, or dissecting microscope clinical research. 5 as it was called back then. The invention of the stereomicroscope at Carl Zeiss was an essential contribution to the rapid upswing in the still young developmental and www.zeiss.de/micro

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The new high-tech microscope procedure, SPIM (Selective Plane Illumination Microscopy), permits fascinating insights into living organisms and makes it possible to observe processes – even those in deep-lying tissue layers. The new development has its roots in the theta microscope from the 1990s which was designed for examinations of large specimens with high 3D resolution. The fundamental light microscopy principle is fluorescence detection at an angle of 90° relative to the illumination axis. SPIM now unites the technology of the targeted illuminated plane in the specimen with the theta principle, thus permitting optical cutting.

SPIM – A New Microscope Procedure

In the SPIM procedure, the specimen is no longer positioned on the microscope slide as usual, but in a liquid- filled specimen chamber which allows it to remain viable during the measurement. Rotating the specimen changes the illumination and detection axes relative to the specimen, permitting better detection of previously hidden or covered areas. Com- plex development processes such as formation of the eyes and brains of fish embryos or other specimens can be observed and documented.

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Jan Huisken, Ernst Stelzer (front) European Molecular and Jim Swoger, Biology Laboratory (EMBL). European Molecular Biology Laboratory (EMBL).

Fig. 1: Medaka fish.

Fig. 2: Pictures of Medaka fish embryos, head region, different perspectives. The last (or fifth) picture 2 in a series shows the fusion of the data sets.

Fig. 3: 180° 90° 3D display of the picture series from Fig. 2. Display in various exposure angles. The center image shows a section through the fusion of the data set.

Fusion

270° 0° 3

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In the SPIM method, which is based nated and observed at a time unlike searchers to delve deeper into the on the theta principle, the specimen in conventional or confocal micro- tissue. The entire process is very fast is illuminated from the side and not scopes in which the entire specimen and the image information generated from above through the objective is exposed for each plane. For exam- can be pieced together using appro- lens as before. With the traditional ple, if 100 planes have to be record- priate software to form a high-reso- configuration, researchers obtained ed, the radiation exposure to the lution 3D image. It is the perfect excellent resolution in the microscope specimen is reduced to 1% of what complement to confocal and multi- slide, but the resolution perpendicu- was previously required. This advan- photon 3D imaging systems. lar to the slide is worse. With the tage can be used to significantly in- SPIM procedure, an extremely thin crease the period of observation. The “light sheet” is generated in the sectional images can be recorded specimen so that an optical sectional from several sides by moving or image is created. A special feature of rotating the specimen. This makes www.embl.de/ExternalInfo/stelzer SPIM is that only one plane is illumi- hidden regions visible, allowing re- www.zeiss.de/micro

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pole yolk cells

somatic cells

y A x 20m 5

SPIM Principle: Camera A beam path and optical components.

pole cells

Tube lens

y B z 6 Detection

Filter

Lightguide Collimator Cylindrical lens Objective Light sheet

from laser Specimen Illumination

Fig. 4: Three-dimensional image rendition of the picture series from Fig. 5: left 180°, right fusion.

Fig. 5: Drosophila embryo, pole cells, x-y plane.

Fig. 6: 7 Drosophila embryo, pole cells, y-z plane.

Fig. 7: Picture series of Medaka fish embryos from various perspectives. (a) (b) (c) (d) 200 m Fig. 8: Drosophila larva: (a) Traditional image (b) Theta illumination of a single plane (c) Image stack (d) Image stack rotated 180° around the vertical axis.

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The Scourge of Back Pain – Treatment Methods and Innovations

Fig. 1: Back pain is among the most com- Dr. Mayer, what developments preserve the mobility and dynamics The ergonomic design of mon health complaints in indus- and changes have you noticed in of the spine, particularly with symp- the surgical microscope allows the surgeon to work trialized countries. More than 30 recent years concerning spine toms of wear. in an extremely comfortable million people in Germany alone diseases? position over longer suffer from chronic or occasional The range of spine diseases has Considering the cost pressures periods. back pain – tense muscles in changed so that now primarily older facing the healthcare sector, what the neck and shoulder regions, people have to undergo surgery on opportunities and innovations are pinched nerves or dislocated ver- the spine. There are many different particularly important from your tebra. types of disease that occur mainly in point of view? Over the long term, spinal pain old age, such as a narrowing of the Any procedure that is considered can considerably affect the quali- spinal canal, degenerative scoliosis minimally invasive is particularly im- ty of life, and not just the elderly and constriction of the nerve canals. portant, i. e. all microsurgical and en- suffer. Signs of wear on the spinal Increasing life expectancy has result- doscopic procedures. In fact, we have column can already be seen from ed in many more patients with symp- been using minimally invasive meth- the age of 30 and onwards. The toms of wear on the spine who ods in Germany for 15 years. Howev- rise in the number of patients require treatment and surgery than er, it was only after healthcare reform with spinal pain and the simulta- in the past. that the positive effects had their full neous cost pressures in healthcare impact. Patients are released earlier are leading to numerous innova- What is the significance of this from the hospital, i. e. the more gen- tions in treatment methods. rise in the number of patients for tle the operation, the shorter the stay Carl Zeiss recently discussed spinal surgery and what are the and the faster the patient can be this with Dr. H. Michael Mayer, consequences? rehabilitated. one of the leading spine surgeons As a result of this development, and medical director at the Or- spinal surgery is becoming more and What is required for minimally thozentrum in Munich. more important, leading to a con- invasive surgery? stant increase in the surgical possibili- There can be no minimally invasive ties. With new techniques, e. g. artifi- surgery without visualization systems. cial discs or minimally invasive sur- Minimally invasive surgery is only gery, it is now possible to intervene possible if optical aids are available, at a much earlier stage with innova- e. g. a surgical microscope. Through tive methods that are designed to tiny incisions, these instruments pro-

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facts

Spinal column

Combined with a complex muscle system the S-shaped spinal column not only provides the body a support beam to walk upright, but also a high degree of mobility and elasticity. The entire spinal column consists of seven cervical, twelve thoracic, five lumbar, five sacral vertebrae and the coccyx con- sisting of three or four vertebrae. The vertebras are separated by discs which work as a buffer, thus enabling mobility of the back. The spinal column also contains a spinal canal in which the very sensitive spinal cord is located. This connects the brain with the body’s organs (peripheral nervous system).

Minimally invasive surgery

Unlike open surgery, minimally invasive techniques avoid using a large incision. Surgical instruments, e. g. a surgical micro- Fig. 2: scope, require a small incision The OPMI® Vario/NC33 of around 2 cm. The new was developed exclusively for minimally invasive techniques deliver a range of procedures on the spine. advantages, including reduced pain, smaller scars and cost savings resulting from shorter hospital stays.

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vide us with the necessary light and very widespread and there is a very magnification – even in the depths of high need for training. the thoracic and lumbar vertebrae. That is what is required. Without it, What can be done so that more you cannot perform minimally inva- doctors, clinics and patients can sive surgery. benefit? It is really just a lot of hard leg- What are the advantages of mini- work. On the one hand, surgeons mally invasive surgery to patients? have to convince other surgeons of We have recorded lower perioper- the benefits of microsurgery and ative morbidity with microsurgery, minimally invasive surgery. This works i. e. as a result of the minimally inva- best in the OR where they can see up sive access, there is less pain and close how this type of surgery works. blood loss, as well as shorter waiting On the other hand, there must be times, hospital stays and rehabilita- courses available, preferably cadaver tion periods. The techniques and in- workshops, in which the participants struments permit surgeons to work can actually practice these techniques much more accurately and safely and see for themselves how advanta- with fewer complications than with geous they really are. I don’t know the naked eye. anyone that has looked through a microscope, or has operated with a Who else benefits from this type microscope, and then later returned of spinal surgery? to traditional surgery. Everyone involved benefits, first and foremost the patient, surgeon How do you think spinal surgery and the assistant. This procedure al- will develop? lows the surgeon to work more safe- Spine surgery is a relatively crisis- ly. Assistants profit because they gen- proof, specialized field which will erally see what the surgeon sees and continue to grow. Within the surgical can learn more easily through onsite disciplines, it is one of the most dy- teaching. Even the entire OR team namically growing specialized fields. benefits, including the nurses, as This can be seen indirectly in the everyone is able to see exactly what growth rates of the medical tech- is happening. Most of all, when you nology industry, the number of spine project the image onto a monitor or implants sold per year and the new the wall everyone is able to see ex- procedures being developed. Further actly what the doctor is doing. There growth is expected, primarily because is really no one who does not benefit more and more surgeons are looking one way or another. to sub-specialize. If you look at the number of knee specialists and hip How common is this type of oper- specialists around the world, then ation? look at the number of spinal column Unfortunately, it is not as common specialists, you can see that there is as we would like. It goes without a significant gap. saying that microsurgery is much more common at neurosurgery cen- What do you envision for the ters than in orthopedic or spine future? trauma centers. Going from the It goes without saying that sur- number of signups for the courses geons want still more flexibility. The that we conduct here in cooperation image of the surgical incision should with Zeiss and the feedback from be visible regardless of the position the many international guests at our of the surgeon’s head or the position hospital, it is obvious that it is not of the monitor.

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It is possible to virtually project facts digitized images using head-mounted displays, in eyeglasses or even in Intervertebral disc space (augmented reality). Augment- ed reality uses certain technology to The intervertebral discs work as a buffer between each create a virtual, 3D image in space. vertebra and maintain their spacing. The plasticity of This technology is currently used pri- the discs enables them to reduce the stress that affects marily in advertising films, but it is the back caused by movement and strong forces. With theoretically possible to use it for increasing age, the fluidity of the disc deteriorates, surgery. thus reducing its elasticity. Nowadays, it is possible to For example, you have a patient implant artificial discs when severe signs of wear ap- and look at the body’s interior pear. Biocompatible disc prostheses implanted between through a certain opening. If you the vertebra help to restore the natural anatomy of could project what you see inside to the body while preserving the mobility of the back. the surface of the body, it would be as if you were working with a normal open wound. You work in the direction you are looking and the interior of the body is projected on the patient’s skin or onto a monitor. This is, of course, something for the future that I think would increase comfort for the surgeon as well as the accept- Constriction ance of minimally invasive surgery. It of the spinal canal would be an ideal surgical technique. Spinal canal stenosis is a narrowing of the spinal canal. Dr. Mayer, thank you very much Various diseases can appear depending on where for an interesting and informative the stenosis is. Most of them are associated with severe interview. pain, organic malfunctions and numbness in the extremities.

Spinal canal stenosis can also be congenital. It can also result from bone diseases, injuries or degenerative changes, through tumors on rare occasions. Discs degenerate considerably faster as the result of improper posture and biomechanical stress. Discs and tissue narrow the spinal canal. Spinal fluid builds up causing painful irritation of the spinal cord.

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ZEISS in the Center for Book Preservation

Books stacked pile-high, plans ence in the area of paper restoration half of the treasures – autographs and files as far as the eye can and combines this knowledge with and books – from the burning see! Diffusing the air with the research into and the development of Unesco world-heritage site. Around delicate mixture of smells – old new methods for preserving large 50,000 volumes from the library, leather, linen, bookbinding glue, quantities of books efficiently and ra- passed from hand to hand, were res- paper, printer’s ink and patina – tionally, a service for which there is cued unscathed in this way. A further that usually greets experts when considerable demand worldwide. 30,000 escaped the inferno, dam- they enter historic libraries and The work of the ZFB is known in- aged to a greater or lesser extent. archives. However, we are not ternationally among experts in this inside one of those venerable old field. In general, however, the center First deep-frozen, buildings; we are actually in an carries out its work and services then dried extremely modern building north largely outside the public domain. and saved of Leipzig, the “Center for Book This changed suddenly in the fall of Preservation” (ZFB). 2004 when the ZFB contributed, us- The latter, already singed by the fire ing methods that it had developed, and soaked by the water used to The ZFB originated from the to saving one of the most valuable extinguish it, were given an initial two central archives, the Deutsche and historically irreplaceable collec- emergency home at the ZFB. Here Bücherei and the Deutsche Biblio- tions of books to have been pre- they were sorted and classified ac- thek, which were amalgamated in served in Germany. What happened? cording to the extent of the damage: Leipzig after the reunification of Ger- On the night of September 2, from Group One, virtually intact, to many. Since 1998, it has offered, as 2004, a devastating fire destroyed Group Six, almost completely de- an independent institution, compre- large parts of the historic Herzogin stroyed. The treatment began with hensive services for the expert preser- Anna Amalia Library building in temporary storage in large cold vation of the valuable collections of Weimar. Residents from that part chambers. Here, wrapped in muslin books held in libraries, archives and of the town, employees and several or fleece, and at a temperature of museums. The center is able to draw hundred spontaneous helpers formed minus 20 degrees Celsius, each soak- on what may well be unique experi- a human chain to save more than ing wet book was transformed within

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a short time into a solid block of ice. ventional sense once it has turned to stalled in rows, ZFB employees turned This prevented any further distortion ice. It ensures that the ice escapes as the pages one at a time and, using and stopped the spread of mold a gas, that is to say, in a dry form. paintbrushes and fine brushes, care- spores; what is more, it allowed A quantity of books weighing up fully removed the mixture of dust the center to gain valuable time. Al- to a tonne is locked in a low-pressure originating from the fire ashes and though the institute worked in three chamber, the internal temperature of lime plaster that the water used to shifts, the careful and expert damage the cooling pipes in the condenser extinguish the fire had washed from limitation process needed time more is lowered to minus 196 degrees the shelves, ceilings and walls of the than anything else. Who would ex- Celsius and the air pressure, which is burning rooms into the books. With pect to be faced with tens of thou- normally around 1,000 millibars, is this after-care process, the ZFB’s res- sands of books, the survival of which reduced to below 6 millibars. Rather cue operation and task were com- hung in the balance from one hour than melting, under these conditions plete. to the next? the ice begins to “evaporate”. In this The patients treated in this way In the second stage of the consistency it can simply be removed have now arrived back in Weimar, process, the books were freeze-dried by suction. Normal air pressure is where the experts and restorers of – a method for which the ZFB has de- restored in the chamber and the the Anna-Amalia library are faced veloped its own system for extracting temperature is gradually raised to with the difficult decision of which the moisture from the books. If they plus 20 degrees Celsius. Depending further rehabilitation measures to were simply left to dry out naturally, on the number and format of the carry out and with what priority. the inks, colors and glues would run. books stored, the treatment process One thing is for sure: it will take The pages would stick together and is often finished after only 3 days or many years and require considerable become distorted and brittle. Addi- fewer. The books are thoroughly financial support before the unique, tional, even more harmful problems dried out. historic, cultural heritage that this would be added to the existing dam- The last stage of the treatment collection represents is open once age. Instead of this, freeze-drying process is the manual removal of again to academics and the public. prevents the moisture in the book any remaining dirt. Below the suction And, even then, the evidence of the block from thawing again in the con- systems of the work cubicles, in- fire will never be completely erased.

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Fig. 1: Acid corrosion, and to improve the glue and quality recording the problem to remedying Damaged books from the the greatest concern Herzogin Anna Amalia of finish. Today it is the acids in the it – to the extent that these measures Library in Weimar. and threat glue which, combined with environ- are possible on the basis of the latest mental influences, destroy the paper. research, findings and techniques. Fig. 2: Mass-deacidification in After this spectacular emergency op- They degrade the fibrous substances This has achieved amazing results – a which the books are eration, the ZFB returned to its “nor- and cellulose which guarantee the scarcely decipherable hand-written soaked in an alkaline, mal fields of activity” in the area of mechanical stability of the books. The musical notation by Beethoven, the non-aqueous solution. book preservation, where rescues fol- The treatment capacity pages become fragile and brittle. This virtually disintegrated first edition of totals over 100 tons a year. lowing fire damage actually, or rather aging process is autocatalytic, mean- a Luther bible as well as the plans for thankfully, form the exception. ing that it accelerates itself. Only a Schinkel building drawn by the ar- It is not bookworms, bark beetles, efficient mass-deacidification is able chitect himself have been protected mold or improper handling that pres- to counteract this deterioration. The against further decay. ent the main threat to books as ZFB has developed what it calls the Naturally, all decisions regarding historic cultural possessions. When Papersave process for this purpose. how and with which methods the asked about the main issues in book During this process, books are satu- various deterioration processes can preservation, Dr. Manfred Anders, rated in an alkaline, non-aqueous so- best be countered are preceded by CEO of the ZFB, named acid corro- lution and deacidified. In this way thorough analyses of the actual and sion, which already threatens a good their life expectancy is extended by a aging condition – using NIR spec- two thirds of all historically and cul- factor of 4 to 5. Although mass- troscopy, for example. It almost goes turally important collections of deacidification (Papersave process) as without saying, therefore, that Zeiss books, newspapers and documents a conservation treatment is able to is present in the Center for Book worldwide, as the greatest problem. delay damage, it is unable to reverse Preservation, acting in some ways as As a result of the growing de- it. In addition, therefore, the ZFB also a partner, with instruments for expert mand for paper, experiments began deals with all kinds of restoration scientific examination, measurement using all kinds of ingredients as early measures: correcting ink corrosion, and determination of methodology. as the 17th and 18th centuries to carrying out paper stabilization, fight- But why go to all this effort when make the diminishing source materi- ing against mold and performing all everything can be recorded on micro- als for paper manufacture go further, forms of damage limitation – from film and digitized, a task that, inci-

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Fig. 3: Aqueous fungicide treatment to kill mildew.

Fig. 4: Removal of ash and lime.

Fig. 5: Paper deterioration caused by acid.

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dentally, is usually carried out in the details ZFB in parallel to the restoration measures? Microscopy

Heritage is an Books have many different ene- leather. Its natural predator is the ongoing duty mies: mice, bookworms, light, book scorpion. micro-organisms and acid. Record- Viewing words, sentences, images ing the damage that a book Discoloration of paper and parch- and drawings on a screen alone is has suffered always forms the ment is usually the sign of an just not the same as still being able starting point of a comprehensive infestation by micro-organisms to hold in your hand the pieces of strategy for book preservation. (mildew, bacteria). These are paper on which, centuries ago, a Microscopes, especially stereo- usually single-cell organisms, good proportion of what forms the microscopes, are frequently used which can be seen under the basis of our history and culture today in the restoration of books to microscope. Dye stuffs, excreted was written down, drawn or printed. analyze the book’s “state of by the micro-organisms, turn In any case, the passage of time and health” before work begins on it: paper green, brown, red, yellow events have already destroyed or a record is made of the book and black, while parchment is ruined much of this. What still re- materials and images of the more likely to show purple marks. mains should not be seen as a prob- damage. These marks remain even after lem that we have inherited but as a the perpetrator has died. How- duty, and the effort of preserving it Liposcelis divinatorius, for exam- ever, colored spots also occur must be regarded as being worth- ple, is a minute, wingless type of if the paper has been attacked by while, even for our own sakes. book louse that lives between the micro-organisms in its structure. pages of books. It prefers to live in a moist, warm environment and feeds on mildew, starch, organic Manfred Schindler glue, fabric, paper, silk and [email protected]

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Across the Globe Russia

Asia North Europe America

Japan China India

Ghana Africa

South America

Australia

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Carl Zeiss Archive Aids Ghanaian Project

Figs 1-5: Without the help of the well-or- A clinic for in fall 2003. The opening of the clin- GansMens Clinic, Kumasi ganized archive system at Carl Ghanaians ic’s own lab followed the start of Fig. 4: Zeiss AG, the laboratory at the operations in early 2005 under the PM2K Photometer “GansMens Clinic” in Kumasi, Albert and Monika Mensah Offei leadership of a general practitioner. Ghana would not yet be com- have been working on the construc- Approximately 30% of all patients plete. An ideal instrument by tion and setup of the GansMens Clin- suffer from malaria. Diarrhea, hepati- Ghanaian standards, an older ic in Kumasi using thier own financial tis, typhoid, diabetes, HIV and other Zeiss photometer – PM2K – was resources since 1997. Albert Mensah infections are also common ailments. available in Vienna, but an Eng- Offei is Ghanaian and decided along In addition to preventive examina- lish version of the user manual with his Austrian wife to create this tions for pregnant women – anemia was not to be found – in Vienna clinic with high quality medical serv- is very common – the clinic has a or any other office. The Carl Zeiss ice, equipment, patient care and small operating room including an in- archives were called on for help. hygiene for his fellow countrymen. It tensive care unit, a children's depart- Dr. Dieter Brocksch, Manager of enables access to medical care, pre- ment (8 beds), 6 patient rooms (20 Technical Information, was able ventive care (e. g. for children, young beds), a maternity ward with a mid- to provide the desired manual in people and pregnant women) and wife and 8 beds. The goal of the a short time. The photometer has the creation of qualified jobs and fur- gynecologist who is also responsible been in use since April 2005 fol- ther education of medical personnel for the maternity ward is to reduce lowing its transport to Ghana in a region lacking sufficient medical the still high mortality rates of both with other medical instruments resources. mother and newborn. Training of shortly before Christmas 2004. The formal requirements for start- high school graduates is done to- up were met with the completion of gether with the Okomfu Anokye construction in 2003 and certification Teaching Hospital in Kumasi. Young

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people who are unable to finance an adapter for use with car batteries ease of use, sturdy and equipped their professional education, particu- for laboratory doctors who work with spare and wear parts, these larly women, use this training to without power. The three pillar strat- “old instruments” have low operat- increase their chances of finding a egy at Med Tech Plus has proven its ing costs and serve well in the mobile job later. value since 1986: recycling and eco- care clinic for rape victims in Bosnia, logical recycling management, mean- at the accident hospital in Timisoar, Recycling medical ingful job training for the long-term in the children’s clinic at the Uni- technology unemployed, repair and re-use to veristy Hospital in Lugansk or in benefit undersupplied regions of the the Health Project in Ngorongoro, All instruments at the GansMens development cooperation. Tanzania – and now at the GansMens Clinic have undergone extensive Instruments that have been sorted Clinic in Kumasi. checks by Med Tech Plus in Vienna. out are collected and repaired. Cari- The procurement of user manuals in tas, Horizont 3000, Ökg, Global the local language is the final link 2000 and Care are among the list of in the chain of adaptation and ad- well-known partners. Many other or- justments that has to be completed ganizations that value robust instru- for the instruments to fit into the ments adapted to the requirements respective local infrastructure and of the respective countries and proj- continue to be used for years to ects have been able to find instru- come. Med Tech Plus used altered ments at Med Tech Plus for their technological solutions to ensure health projects in Nicaragua, Cuba, sustainability (e. g. a microscope with Peru, Uganda, Kenya, Nigeria, Alba- Peter Gluchi, Med Tech Plus, Vienna. a mirror for sunlight illumination and nia and the Ukraine. Adjusted for www.medtechplus.at

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Prizes and Awards

100 Years of Brock& Michelsen

Carl Zeiss Exclusive Partner

Fig. 1: On April 1, 1905, a 20-year-old sales- Knud Michelsen and man, Knud Michelsen, and a 22- Peter Brock in 1953 looking through a standard year-old precision mechanic, Peter microscope. Brock, opened P. Brock & Co. in Copenhagen. The company later Fig. 2: Festivities for the became Brock & Michelsen Opto- 100th anniversary mechanical Institute and Workshop. (left to right: Dr. Norbert Having early contacts with Carl Zeiss Gorny, Gregers Brock and Jørgen Brock). in Jena, Brock & Michelsen became the company’s general representative in Denmark in 1921, making 2 Brock & Michelsen Carl Zeiss’ longest-serving active representative outside Germany. Approximately 60 employees support products ranging from microscopes and the various medical systems to products from semiconductor production technol- Helsinki ogy. Oslo Tallinn Stockholm

www.brockmichelsen.dk www.zeiss.dk Sweden

Riga Århus

Dublin Copenhagen

Vilnius

48 Innovation 15, Carl Zeiss AG, 2005

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Award for NaT Working Project

details The 4th NaT Working Symposium took place at the German Cancer Research Center in Heidelberg NaT Working from February 27 to March 1, 2005. The Robert Bosch Foundation initiated NaT Working. The program is intended to arouse students’ interest in Five awards were distributed among natural sciences and technology. One promising way the 60 sponsored projects. The NaT of accomplishing this is to establish and maintain Working Project, “I see what you personal partnerships between teachers, students and don’t see: small and large on an ex- scientists and engineers active in research. Internships pedition to the microcosm” from for students and teachers in the researchers’ labs, Birger Neuhaus and his team from summer schools, student congresses or game-like the Carl Zeiss Microscopy Center practical projects during leisure time are among the from the Humboldt Exploratorium at sponsored activities. Particularly outstanding projects the Museum of Natural History in are presented with an award once a year. Berlin finished second and received an award of 4,000 euros. www.bosch-stiftung.de/natworking

Fig. 1: The overjoyed winner with Dr. Ingrid Wünning from the Robert Bosch Foundation, and Karsten Humboldt Exploratorium Berlin Schwanke, meteorologist and television presenter. Exploring nature and discovering new things is the Fig. 2: greatest motivation for scientists. The Humboldt At the exhibition booth. Exploratorium offers young people the opportunity of participating in the joy of scientific discovery. The Exploratorium and Humboldt University in 1 Berlin owe their name to Alexander von Humboldt – a multi-talented natural scientist who was a geologist, zoologist and botanist in one – and his brother Wilhelm.

www.humboldt-exploratorium.de www.naturkundemuseum-berlin.de

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Product Report

Digital Pathology: Quality and efficiency Immuno-histological specimens are MIRAX SCAN of the diagnosis digitized before the specimens fade. Several specimens can be observed at The demands on medical diagnostics MIRAX SCAN unites optics and tech- the same time, current and previous are continually increasing. Whether nology from Carl Zeiss with IBM’s ex- sections can be compared quickly and clinical labs, research service providers perience in digital archiving. The inte- directly. The diagnosis process can or pharmaceutical companies, time grated systems solution for digital also be documented on the section and cost pressures are high. Cus- pathology enables absolute concen- and can be traced at any time. tomers want maximum quality and tration on the essentials – the diagno- All archive data is directly available on the network: a real time savings growing efficiency. A large number sis. High-resolution digital data sets, MIRAX SCAN of external factors impacts work digital slides, enhance the quality and during complex evaluation processes processes in pathology today: compe- efficiency of the evaluation of the and when comparing different speci- tition amongst clinical facilities has findings. MIRAX SCAN automatically mens. The pathological findings can long been a part of the picture. generates digital slides for up to 300 be immediately integrated into the Borderline cases often lead to legal specimens in one run – even in un- patient’s electronic file and quickly disputes. Everyday clinical life must interrupted overnight operation. accessed. ensure compliance with legal guide- The diagnosis and report are right on As soon as the section specimens are lines. Health authorities are increas- the monitor which does not have available, MIRAX SCAN does the rest ingly demanding more from pharma- to be at the same location as the with its powerful optics. The results ceutical research. Lab standards are scanner. The data and results are can be seen on the monitor with the becoming more challenging. To top transmitted directly via the internal same excellence as through the mi- it off, the amount of work is also network or the Internet. The sections croscope. increasing. High throughput is no can be seen as a whole. longer the exception. Time and technology are always in high demand.

UHRTEM Superlux™ Eye the retina must not be too strong. Xenon Illumination It can lead to tissue damage. Carl UHRTEM, the latest generation of Zeiss rose to the challenge and opti- ultra-high resolution transmission With the innovative Superlux™ Eye mized xenon for ophthalmology. With electron microscopes from Carl Zeiss, xenon illumination, Carl Zeiss provides Superlux™ Eye illumination, the UV made the breakthrough in sub- ophthalmic surgeons with a white portion below 408nm is filtered, Angstrom image resolution (0.8 light that meets even the highest de- making it just as safe as halogen Angstroms, or 0.08 mm). This mile- mands. This clearly improves working illumination. Compared to halogen, stone was achieved using the newly conditions for ophthalmic surgeons: however, Superlux™ Eye has more developed 200 kV field emission more realistic colors, increased con- advantages: it has a significantly UHRTEM. The instrument developed trast, better detail recognition and lower IR portion which reduces the in partial cooperation with CEOS enhanced quality of videos resulting thermal exposure to the cornea and GmbH in Heidelberg is equipped from the higher color resolution. other tissues, making operations on with electron-optical components to Compared with standard halogen the eye even more safe and gentle. correct aberrations, electron-beam illumination, Superlux™ Eye is also monochromatization and energy-fil- easier to service. tered imaging. UHRTEM is a special Xenon illumination is nothing new www-zeiss.de development for the sub-Angstrom to surgical microscopes in other disci- characterization of advanced materi- plines – neurosurgery and ENT sur- als and module structures from inno- gery, for example. However, until vative nanotechnology. now, it was considered too dangerous www.smt.zeiss.com for ophthalmology as the light hitting Sub-Angstrom UHRTEM with revolutionary suspension on the microscope column.

Young’s edge zone sample provides an insight into the usability of image resolution of 0.8 Angstroms. An area of the image section is displayed at a resolution of 0.7 Angstroms. OPMI® VISU 210

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Masthead Innovation, The Magazine from Carl Zeiss No. 15, July 2005 Publishers: Carl Zeiss AG, Oberkochen Corporate Communication Marc Cyrus Vogel. Editors: Dr. Dieter Brocksch, Carl Zeiss AG 73446 Oberkochen, Germany Phone +49 (0)7364 203408 Fax +49 (0)7364 203370 [email protected] Gudrun Vogel, Carl Zeiss Jena GmbH 07740 Jena, Germany Phone +49 (0)3641 642770 Fax +49 (0)3641 642941 [email protected] English version of the magazine: Translation Service (S-KS), Carl Zeiss AG, Oberkochen Authors at Carl Zeiss: [email protected] www.zeiss.de Other authors: If no information is given to the contrary, they can be contacted via the editor. If readers have any inquiries about how the magazine can be obtained or if they wish to change their address (the customer number should be indicated, if applicable), we would kindly ask Carl Zeiss Optics in users of high-end cell phones to them to contact the editor. Nokia Mobile Phones record, save, show and print images Cover photo: and videos with even better quality. Archives of Carl Zeiss AG, Photolab EMBL Carl Zeiss has supported Nokia, the The Nokia N90 is the first product Heidelberg, Andreij Popov EMBL global market leader in mobile com- with integrated ZEISS optics from Hamburg, Robert Koch Institute. munications, since the end of April this partnership: it is a mobile phone Idea, conception, implementation of 2005, with the integration of Carl with a 2 megapixel camera and the cover photo: Dieter Brocksch, Andreas Zeiss optics into Nokia camera smallest Tessar lens in the world. Schwab (Carl Zeiss AG, S-K), phones. In the future, this will enable Nicola Schindler, Annette Jenak, Elisabeth Jenewein (MSW Aalen). Photo on page 41: gettyimages. Photos on pages 42-45: With the kind permission of Zentrum für Bucherhaltung GmbH. Photos on page 49: With the kind permission of the Robert Bosch Foundation/Jyan de Andres. Design: Corporate Design, Carl Zeiss AG, 73446 Oberkochen. Layout and composition: MSW, 73431 Aalen, www.msw.de. Printed in Germany by: C. Maurer, Druck und Verlag, 73312 Geislingen a. d. Steige. ISSN 1431-8059 © 2005, Carl Zeiss AG, Oberkochen. Permission for the reproduction of individual articles and pictures – with appropriate reference to the source – will gladly be granted after prior consultation with the editors. Picture sources: Unless otherwise speci- fied, all photographs were contributed by the authors or originate in the Carl Zeiss archives. www.zeiss.de/photo Articles in which the author’s name has www.nokia.com been given do not necessarily reflect the opinion of the editors.

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InnovationThe Magazine from Carl Zeiss