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Rockefeller University Digital Commons @ RU The Rockefeller Institute Quarterly The Rockefeller University Newsletters 12-1962 The Rockefeller Institute Quarterly 1962, vol. 6, no. 2 The Rockefeller University Follow this and additional works at: http://digitalcommons.rockefeller.edu/ rockefeller_institute_quarterly Recommended Citation The Rockefeller University, "The Rockefeller Institute Quarterly 1962, vol. 6, no. 2" (1962). The Rockefeller Institute Quarterly. Book 22. http://digitalcommons.rockefeller.edu/rockefeller_institute_quarterly/22 This Book is brought to you for free and open access by the The Rockefeller University Newsletters at Digital Commons @ RU. It has been accepted for inclusion in The Rockefeller Institute Quarterly by an authorized administrator of Digital Commons @ RU. For more information, please contact [email protected]. 1 THE DECEMBER * 1962 THE ROCKEFELLER INSTITUTE A GRADUATE UNIVERSITY AND RESEARCH CENTER \ Electron Microscopy at the Institute BY RAY M 0 N D B . G R I F F I T H S Executive Editor of 'The Journal of Cell Biology THE DEVELOPMENT within the past sented to Claude in 1943 by Albert E. the ground substance, nucleus and mito- two decades of the electron microscope Gessler, then Director of Research at the chondria. Distortion was not excessive de- and of techniques for using it to study Interchemical Corporation, New York, spite the extreme dryness and thinness of biological structure has ushered in an where a research laboratory was equipped the specimens, and details were revealed epoch of new discoveries. With the high with one of the earliest models of the elec- of submicroscopic structure never before resolving power of this instrument, which tron microscope. In his first studies, car- seen. The paper noted that osmium tetrox- is over one hundred times greater than ried out with Ernest E Fullam, an engi- ide fixation gave the most truthful images that of the light microscope, biologists neer at the Interchemical Corporation, and that in the best osmium preparations have explored the architecture of the cell Claude succeeded in obtaining electron the ground substance of the cytoplasm in the submicroscopic realm (zooo A micrographs of fractionated material had a "lace-like reticulum" with "vesicle- down to zo A and below) and have found sprayed onto the specimen'grid of the like bodies, i.e., elements presenting a this to consist of a richly differentiated microscope, but the resulting images were center of less density, and ranging in size fine structure of great complexity. From not satisfactory enough for an accurate in- from loo to 150 m~ . arranged along the the systematic study of the fine structural terpretation of structural details. Though strands of reticulum." This discovery, evi- organization of cells, new concepts of mor- this preliminary work yielded meager in- dence of "invisible" organization in the phology have emerged, and these bear on formation on fine structure, it did indicate cell ground- substance, was to have im- the fundamental principles of all biologi- the potential value of electron microscopy portant implications for the future course cal disciplines. Such studies, together and also defined some of the basic re- of cell research. with those employing biochemical tech- quirements for the preparation of speci- In 1945 an electron microscope ob- niques in the analysis of the functional ac- mens. tained by The Rockefeller- Foundation tivities of cell components, have begun to was set up in the Institute and was made CULTUREDCELLS elucidate basic structural-functional rela- available to Claude and Porter for their tions within cells and even to make possi- One of the most important considera- continuing work on cultured cells. In one ble their interpretation in molecular terms. tions was that specimens should be thin. of their studies, carried out in collabora- At The Rockefeller Institute pioneering It occurred to these investigators that tis- tion with Edward G. Pickels, Claude and ' efforts in the biological exploitation of sue culture cells which spread out exten- Porter examined cultured chicken tumor electron microscopy were made in the sively when grown on a flat surface might cells to determine the nature of the filtra- early 1940's by Albert Claude and Keith be thin enough for the differential scat- ble agents responsible for transmission of R. Porter, then working in the Laboratory tering of electrons and the formation of these tumors to new hosts. Previous filtra- of Cancer Research under J. B. Murphy. useful images. Keith Porter, who was tion studies had indicated the transmit- * Over the years this work has prospered, working with cultured tissue cells, collab- ting agent to be particulate and of small and one result has been the growth of a orated with Claude and Fullam in trial size, but confirmation of its character had vigorous school of cytology. Much of the studies. In preparing specimens for exam- been unobtainable by the limited resolu- technique evolved within this school has ination, Porter employed a fixation with tions of the light microscope. Using the now been adopted in other laboratories, osmium tetroxide and worked out an in- new techniques, these investigators were and by other groups at The Rockefeller geniously simple method for transferring able to demonstrate in the tumor cells the Institute. The historical account which the cultured cells from coverglasses, on presence and distribution of "small bod- follows is concerned only with the work which they had grown, to the specismen ies" of the size estimated for the tranmit- of the school of cytology and its develop- grid of the microscope. Both the selection ting agents. From this work came the first ment since the early forties. of specimens and the method of their microscopic description of the Rous sar- preparation proved to be favorable, as coma virus. CELL FRACTIONS electron micrographs were produced in For some time Claude had been devel- which clear images of cell structures were MICROTOMY oping techniques for sLparating cell com- discernible. By the late forties it had been amply ponents by differential centrifugation and The results of the work of Porter, demonstrated that the electron micro- had isolated from homogenates of liver Claude and Fullam were reported in 1945, scope was an effective means of examin- and other tissues a mitochondria1 fraction and their paper was the first demonstra- ing cells grown in uitro, but its application and a "small granule" (later called micro- tion that intact cells could be examined to the study of cells in situ, in their normal somal) fraction. The opportunity to ex- by electron microscopy. The electron mi- surroundings within tissues, was ham- amine his fractions with the high reso- crographs revealed all the cellular entities pered by the difficulty of preparing thin lutions of electron microscopy was pre- known from light microscopy including sections. Claude and Fullam, like investi- gators at other laboratories, turned their The year 1949 was an eventful one for by poor preservation. Palade first studied attention to the problems of microtomy. the future development of electron mi- the fixation of cell preparations with 0s- With Joseph Blum, a talented instrument croscopy at the Institute. Knowing the mium tetroxide, and defined the optimum maker at the Institute, Claude developed progress already made, and being con- conditions for using it. As a resilt he was " . a smgle-pass" microtome in which a vinced of the importance of the method in able to resolve in fine detail the structure specimen was fixed to the edge of a re- cell research, Dr. Herbert S. Gasser, then and spatial relationship of cell compo- volving disc or wheel and thus carried Director of the Institute, established a nents in thin sections of cells. Palade used past a knife, but this was not entirely sat- Cytology Laboratory and appointed Keith his improved technique for the study of isfactory. Eventually the chief contribu- Porter to direct its work. Two rooms in mitochondria. He examined the mitochon- tion to the technique of microtomy the basement of Theobald Smith Hall dria in the cells of a variety of mammalian emerged in the form of the Porter-Blum were designated for the laboratory and tissues and found that they have a charac- instrument, in which by mechanical con- were fitted very soon with a new RCA teristic pattern. Each mitochondrion was trol a specimen is brought against a knife. electron microscope, purchased by the observed to possess an outer limiting This device has now become the standard Institute, and the "old" RCA instrument membrane and a second inner membrane microtome for electron microscopy. bought by the Foundation. Within the from which a system of regularly spaced While his associates were attempting year, Porter was joined by George E. ridges (which he termed cristae mito- to prepare tissues for electron microscopy, Palade, a colleague from, J. B. Murphy's chondriales) projects into a structureless Porter continued to utilize cultured cells Laboratory, who had been conducting matrix. In the most favorable micrographs, in his work and, with the assistance of cell fractionation studies in collaboration the cristae appeared as folds of the inner his students, he examined the fine struc- with Claude and with Walter C. Schnei- membrane. Palade interpreted these struc- ture of various cultured materials, includ- der and the late George H. Hogeboom. tural details as the expression of a more ing tumor cells, blood clots, fibroblasts, Thus began the association between Por- complex organization at the molecular and other cell types. He demonstrated the ter and Palade that fostered and gave level. In correlating his findings with the virus-like particulate bodies (thought to leadership to the development of elec- known biochemical data, he postulated be milk agent) associated with the epi- tron microscopic research in cytology that the framework of the internal mem- thelial cells of transmissible mammary throughout the world.

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