DOCSLIB.ORG

PREPARATION of DECAYED WOOD for MICROSCOPICAL EXAMINATION Table of Contents

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
PREPARATION of DECAYED WOOD for MICROSCOPICAL EXAMINATION Table of Contents U.S. DEPARTMENT OF AGRICULTURE • FOREST SERVICE • FOREST PRODUCTS LABORATORY l MADlSON, WIS U. S. FOREST SERVICE RESEARCH NOTE FPL-056 August 1964 PREPARATION OF DECAYED WOOD FOR MICROSCOPICAL EXAMINATION Table of Contents Page Summary . 1 Introduction . 1 Preparation of Decayed Samples . 2 Embedding Methods ............................. 2 Celloidin ................................ 2 Paraffin ................................ 5 Polyethylene Glycol .......................... 6 Freezing. ............................... 7 Maceration. 8 Sectioning . 8 Preparation of Sections for Staining. .................... 10 Adhesives ............................... 10 Removal of Embedding Matrix. .................... 11 Staining. ................................... 11 Differentiation of Hyphae and Wood .................. 12 Picro aniline blue ........................ 12 Pianeze IIIb ........................... 13 Differentiation of Wood Structures and Components ......... 13 Safranin and fast green ..................... 14 AzureB ............................. 15 Zinc-chlor-iodide and phloroglucinol .............. 15 Iodine-potassium iodide ..................... 16 Mounting. 17 Microscopical Methods ........................... 18 Observational ............................. 18 Measurement ............................. 18 Literature Cited . 21 FPL-056 PREPARATION OF DECAYED WOOD FOR MICROSCOPICAL EXAMINATION By W. WAYNE WILCOX, Pathologist Forest Products Laboratory,’ Forest Service U.S. Department of Agriculture ------ Summary This report describes some of the methods that were devised or found to be particularly satisfactory for the microscopical observation of wood in various stages of decay. These methods include the celloidin, paraffin, and polyethylene glycol embedding methods, methods for macerating, sectioning, staining, and mounting, and a discussion of several optical systems which facilitate micro- scopical observation of decayed wood. A rapid method for the measurement of changes in the amount of cell wall substance visible in cross sections is discussed. Introduction Because of its generally soft or friable nature, decayed wood may be difficult to prepare satisfactorily for microscopical examination. For example, thin sectioning often requires that an embedding matrix be used to hold the wood structure intact during the cutting process. Since samples of decayed wood may differ greatly in their hardness and strength, it is desirable to have available a number of methods, each of which may be applicable to a specific set of con- ditions. This report is a survey of some methods used in a detailed study of changes caused by decay in the microstructure of wood, which was undertaken in the preparation of a Doctoral thesis. 1 Maintained at Madison, Wis., in cooperation with the University of Wisconsin. FPL-056 Preparation of Decayed Samples In this study, suitable decay samples were obtained by following the major procedures of the soil-block method outlined in ASTM Standard Method D1413-61 (1).2 Blocks to be decayed were cut with a thickness of 1/8 inch along the grain, so that specimens of satisfactory size for microscopical exam- ination could be prepared by simply splitting the blocks along the grain with a sharp knife. Reducing the thickness of the blocks increased the uniformity of decay and made further cutting across the grain unnecessary. Cutting across the grain, after the blocks had been decayed, could have produced considerable dis- tortion. The blocks were conditioned and weighed according to the specifications of the soil-block method.. Following incubation the blocks were again conditioned and weighed to determine the weight loss that had occurred. Small specimens were then split from the blocks and embedded. Although air-drying of the blocks had no apparent adverse effects upon wood structure, hyphae present in the wood collapsed and became distorted. Presumably this difficulty could be avoided by submerging the blocks in a fixative (7 ,15) im- mediately after removal from culture, but such treatment would prevent accurate determination of the weight loss sustained by the blocks. Embedding Methods Celloidin The celloidin method proved to be the most satisfactory procedure for embedding decayed wood. The results fully justified the required expenditure of time--nearly 2 months for the preparation of fully embedded specimens. Even the slight amount of structure still present in white-rotted wood at weight losses of over 70 percent was held intact by this method during the cutting of sections 4 microns in thickness. The embedding of specimens in celloidin required no special skills but entailed following a number of steps with reasonable care. The specimens were first either air-dried or dehydrated by an ethyl alcohol series (15). The dry specimens were placed under vacuum in several changes of absolute ethyl alcohol until they 2 Underlined numbers in parentheses refer to Literature Cited at the end of this report. FPL-056 -2- absorbed enough alcohol and sank. They were then transferred to several changes of ethylene glycol monomethyl ether (also under vacuum), which proved to be an excellent solvent for celloidin. Unlike ether-alcohol, it presents no explosion hazard and it evaporates slowly enough to allow convenient handling. The speci- mens in ethylene glycol monomethyl ether were placed in a stoppered bottle in an oven at 52° to 54° C. for 2 days or more. They were transferred successively through 2, 4, 6, 8, and 10 percent3 solutions of celloidin in ethylene glycol mono- methyl ether maintained at 52° to 54° C., and were allowed to remain in each grade for a minimum of 2 days. With a film of celloidin around them at all times, the specimens were removed from 10 percent celloidin, transferred with the face to be sectioned downward, to hardening chambers (fig. 1),4 and covered with a 12 percent solution of celloidin in ethylene glycol monomethyl ether. The celloidin in the hardening chambers was allowed to concentrate by evaporation at room temperature. Additional celloidin was added periodically in order to keep the chambers full and to allow for further concentration. During this process, the surface of the celloidin became dry and hard. The configuration of the surface crust was used as an indicator of the concentration of the celloidin to be added. Thus, when the crust of celloidin formed a ring around the edge of the chamber, a 20 percent solution of celloidin in ethylene glycol monomethyl ether was added, but when a solid plug of hard celloidin formed at the mouth of the chamber a 40 percent solution was added. The crust was removed with a knife prior to each addition to prevent the trapping of air bubbles in the matrix. The 40 percent celloidin was allowed to evaporate at room temperature until it became quite firm, or until it, began to pull the bottom of the chamber inward. At this stage there was little danger of the celloidin becoming too hard, provided the shrinkage did not begin to distort the specimen, since it was found that very hard celloidin still could be successfully sectioned merely by increasing the period of storage in glycerin-alcohol. The embedded specimen was next removed from the hardening chamber. With a knife, the chamber was cut away from the glass slide and loosened from its contents. The embedded specimen was then removed from the chamber by pressing on one end and placed on a wooden mounting block of such size that it could be firmly held in the microtome. The mounting block had been impregnated previously with 15 to 25 percent celloidin in ethylene glycol monomethyl ether. The embedded specimen was then surrounded with 40 percent celloidin and sub- merged in chloroform overnight, or until completely hardened throughout. 3 Throughout this report, the term “percent.” as applied to solutions of solid reagents, denotes the number of grams of solute per 100 milliliters of solvent. The percentage concentration of liquid reagents is expressed on a volumetric basis. 4 The idea from which these chambers were developed was originally that of Dr. Catherine Duncan of the Forest Products Laboratory. FPL-056 -3- Figure 1.--Celloidin hardening chambers. These consisted of two 1/2-inch-long segments of plastic tubing 1/2 inch in outside diameter adhered to a 1- by 3-inch glass microscope slide with a room-temperature-curing nitrile-phenolic resin in organic solvents. The use of a glass slide with one etched end allowed the contents of the chambers to be labeled with a pencil. Such chambers facilitated the handling of large numbers of specimens and allowed the specimens to be oriented for sectioning at a stage in the embedding process when they could be easily observed. M 126 011 FPL-056 -4- After removal from the chloroform, the hardened material was trimmed down for sectioning and stored in a 50-50 mixture of glycerin and 95 percent ethanol. The longer the material was stored in glycerin-alcohol, the better were its cutting properties. Sectioning was performed on a sliding microtome. Paraffin Paraffin embedding was compared with celloidin for specimens in very advanced stages of decay, since with paraffin there would be less opportunity for dis- tortion or damage due to handling. Paraffin proved quite satisfactory for material in advanced stages of decay but was inferior to celloidin embedding for sound wood or wood in early stages of decay. An advantage of paraffin embedding was that serial sections could be obtained by sectioning on a rotary microtome. The methods of Sass (15) for paraffin embedding,
Load more

Recommended publications
  • Gst Gram Staining Learning Objectives the Student Will  Use Aseptic Techniques in the Safe Inoculation of Various Forms of Media
    GSt Gram Staining Learning Objectives The student will Use aseptic techniques in the safe inoculation of various forms of media. Follow oral and written instructions and manage time in the lab efficiently. Use the bright field light microscope to view microbes under oil immersion, make accurate observations and appropriate interpretations and store the microscope according to lab procedures. Properly prepare a bacterial smear for accurate staining and describe the chemical basis for simple staining and negative staining. Background/Theory Differential staining distinguishes organisms based on their interactions with multiple stains. In other words, two organisms may appear to be different colors. Differential staining techniques commonly used in clinical settings include Gram staining, acid-fast staining, endospore staining, flagella staining, and capsule staining. This link to the OpenStax Microbiology text provides more detail on these differential staining techniques. (OpenStax CNX, 2018) The Gram stain is a differential staining procedure that involves multiple steps. It was developed by Danish microbiologist Hans Christian Gram in 1884 as an effective method to distinguish between bacteria containing the two most common types of cell walls. (OpenStax CNX, 2018) One type consists of an inner plasma membrane and a thick outer layer of peptidoglycan. The other type consists of a double phospholipid Figure 1 Simplified structures of Gram negative cells (left) and Gram positive bilayer with a thin layer of cells (right) peptidoglycan between the two. The Gram Staining technique remains one of the most frequently used staining techniques. The steps of the Gram stain procedure are listed below and illustrated in Figure. (OpenStax CNX, 2018) 1.
    [Show full text]
  • The Suitability of Certain Stains 3R Studying Lignification in Balsam Fir, Ibies Balsamea (L.) Mill
    The suitability of certain stains 3r studying lignification in balsam fir, ibies balsamea (L.) Mill Kutscha and Gray Technical bulletin 53 March 1972 Life Sciences and Agriculture Experiment Station Cover photo: Safranin and aniline blue, showing blue unlignified cambium (top of photo) and contrasting red lignified tissue (lower part of photo). Imma­ ture, secondary walls appear blue-blue green and can be seen approximately five to eight cells down from the cambial region. Section of FAA-killed and celloidm embedded compression wood sample collected July 6, 1966; X 320. ABSTRACT An investigation was conducted to examine the suitability of ten staining reactions for studying lignification in balsam fir, Abies balsamea (L.) Mill. Two experiments were carried out on material collected on two different dates. In each experiment slides of fresh, FAA-killed and FAA-killed celloldin-embedded material of normal and compression wood were stained and evaluated. No significant difference in staining reactions was found between material collected on different dates. In each experiment, the embedded material showed somewhat superior results compared with the fresh and FAA-killed material with at least half of the stains. No marked difference was observed between normal and compression wood. This study emphasized the need for considsring each of the ten staining reactions on an individual basis, since each has particular ad­ vantages and disadvantages as emphasized throughout the study. Stain­ ing schedules were prepared and tables compiled to determine
    [Show full text]
  • Revisions Inserts Rev from Rev to JOB
    BALTSO0191 Version 11.0 Template 4 Revisions Inserts Rev from Rev to JOB # 06 07 52-17 Notes: 1. BD Catalog Number: 212525, 212526, 212527, 212528, 212531, 212532, 212539, 212542, 212543, 212544, 212545 2. Blank (Sheet) Size: Length: 25.5” Width: 22” 3. Number of Pages: 28 Number of Sheets: 1 4. Page Size: Length: 8.5” Width: 5.5” Final Folded Size: 4.25” x 5.5” 5. Ink Colors: No. of Colors: 2 PMS#: 032 Red; Standard Black 6. Printed two sides: Yes X No 7. Style (see illustrations below): # 5 W W W W W W W 8. Vendor Printed X Online/In House Printed Web 9. See specication control no. N/A for material information. 10. Graphics are approved by Becton, Dickinson and Company. Supplier has the responsibility for using the most current approved revision level. Label Design COMPANY CONFIDENTIAL. THIS DOCUMENT IS THE PROPERTY OF BECTON, DICKINSON AND Becton, Dickinson and Company Proofer COMPANY AND IS NOT TO BE USED OUTSIDE THE COMPANY WITHOUT WRITTEN PERMISSION. 7 Loveton Circle Sparks, MD 21152 USA Checked By Category and Description Sheet: 1 of 29 Part Number: Package Insert, 8820191JAA Gram Stain Kits and Reagents Scale: N/A A B Gram Stain Kits and Reagents English: pages 1 – 5 Italiano: pagine 14 – 18 8820191JAA(07) Français : pages 5 – 9 Español: páginas 19 – 23 2017-09 Deutsch: Seiten 10 – 14 Contact your local BD representative for instructions. / Свържете се с местния представител на BD за инструкзии. / Pokyny vám poskytne místní zástupce společnosti BD. / Kontakt den lokale BD repræsentant for at få instruktioner.
    [Show full text]
  • GRAM STAIN REAGENTS - for in Vitro Use Only - Catalogue No
    GRAM STAIN REAGENTS - For in vitro use only - Catalogue No. SG51-55 Our Gram-Stain Reagents are intended to be The last step is the application of a counterstain. The used as a differential stain for the microscopic most common counterstain is safranin, which colors examination of bacterial cultures and laboratory decolorized cells pink. An alternate counterstain is basic specimens. fuchsin, which gives the decolorized cells more of a Gram staining is the single most useful test in bright pink or fuchsia coloration. The basic fuchsin the microbiology laboratory given its simplicity and counterstain works particularly well for anaerobic ability to differentiate bacteria into two main bacteria, but poorly for Legionella and Bordetella groups: gram-positive organisms and gram negative species. organisms. Hans Christian Joachim Gram first devised the original procedure in the late 19 th century, and although modifications have since been Formulation per Litre made, the basic principles and results remain the same. Our formulation is often referred to as the SG51 Gram Crystal Violet Hucker Modification. The staining spectrum includes almost all Crystal Violet ................................................ 20.0 g bacteria, many fungi, and parasites such as Ammonium Oxalate ....................................... 8.0 g Trichomonas , Strongyloides , and protozoan cysts. Methanol ................................................. 200.0 mL The notable exceptions include intracellular pathogens such as Chlamydia and Rickettsia , and SG52 Gram Iodine those organisms lacking a true cell wall such as Mycobacterium , Mycoplasma , and Ureaplasma . Iodine Crystals .............................................. 3.33 g The differential properties of the staining Potassium Iodide ........................................... 6.67 g process are attributed to the differences in composition between gram-positive and gram- SG53 Gram Decolorizer negative cell walls.
    [Show full text]
  • How to Construct and Use a Simple Device to Prevent the Formation of Precipitates When Using Sudan Black B for Histology
    Acta Botanica Brasilica 29(4): 489-498. 2015. doi: 10.1590/0102-33062015abb0093 How to construct and use a simple device to prevent the formation of precipitates when using Sudan Black B for histology João Marcelo Santos de Oliveira1 Received: April 17, 2015. Accepted: July 1, 2015 ABSTRACT The present work aims to demonstrate the stages of fabrication and use of a simple device to avoid the formation or fixa- tion of precipitates from Sudan Black B solution on tissues. The device consists of four coverslip fragments attached to a histology slide, which serve as points of support for the histological slide under analysis. To work properly, the histology slide with the sections should be placed with the sections facing downwards the device. A small space between the device and the histology slide is thereby created by the height of the coverslip fragments. When Sudan Black B is applied, the solution is maintained within the edges of the device and evaporation is minimized by the small space, thereby reducing the consequent formation of precipitates. Furthermore, by placing the sections facing downward the device, any sporadically formed precipitates are prevented from settling on and fixing to the sectioned tissues or organs. By avoiding the formation of precipitates, plant cells, tissues and organs can be better observed, diagnosed and photomicrographically recorded. Keywords: histochemical tests, histology, lipids, plant anatomy, Sudan Black B Introduction for organic solvents, printer ink, varnishes, resins, oils, fats, waxes, cosmetics and contact lenses. Sudan reagents, including the traditional Sudan III, IV Lansink (1968) isolated two pure fractions of Sudan and Sudan Black B (SBB), are widely utilized to determine Black B, in addition to impurities, and denominated them lipids (Horobin 2002) in animals, plants and hydrophobic SBB-I and SBB-II.
    [Show full text]
  • DIFFERENTIAL STAINING, Part I
    DIFFERENTIAL STAINING, Part I Differential staining is a procedure that takes advantage of differences in the physical and chemical properties of different groups of bacteria. It allows us to differentiate between different kinds of bacterial cells or different parts of a bacterial cell. I. GRAM STAIN The most commonly used differential stain is the Gram stain, first described in 1884 by Christian Gram, a Danish physician. The Gram reaction divides bacteria into two groups, those which are Gram-positive and those which are Gram-negative. Those organisms which retain the primary stain (crystal violet) are stained purple and are designated Gram-positive; those which lose the crystal violet and are subsequently stained by a safranin counterstain appear red and are designated Gram-negative. The conventional Gram-stain technique is described in the Procedure part of this handout; however, it is important to recognize early on that two aspects of the procedure are crucial: 1. The crystal violet treatment must precede iodine treatment. Iodine acts as a mordant, i.e., it increases the affinity of the cells for the crystal violet. Iodine alone has no bacterial staining capabilities. 2. Decolorization must be short and precise. Too long an exposure to 95% alcohol will decolorize Gram-positive as well as Gram-negative cells. The Gram stain has been used as a taxonomic tool for many years, aiding in the classification and identification of bacterial cells. However, it is also useful in a broader sense, as there appears to be a close correlation between the Gram reaction and many other morphological and physiological characteristics of bacterial cells.
    [Show full text]
  • STAINING TECHNIQUES — Staining Is an Auxiliary Technique Used in Microscopy to Enhance Contrast in the Microscopic Image
    STAINING TECHNIQUES Staining is an auxiliary technique used in microscopy to enhance contrast in the microscopic image. Stains or dyes are used in biology and medicine to highlight structures in biological tissues for viewing with microscope. Cell staining is a technique that can be used to better visualize cells and cell components under a microscope. Using different stains, it is possible to stain preferentially certain cell components, such as a nucleus or a cell wall, or the entire cell. Most stains can be used on fixed, or non-living cells, while only some can be used on living cells; some stains can be used on either living or non-living cells. In biochemistry, staining involves adding a class specific (DNA, lipids, proteins or carbohydrates) dye to a substrate to qualify or quantify the presence of a specific compound. Staining and fluorescence tagging can serve similar purposes Purposes of Staining The most basic reason that cells are stained is to enhance visualization of the cell or certain cellular components under a microscope. Cells may also be stained to highlight metabolic processes or to differentiate between live and dead cells in a sample. Cells may also be enumerated by staining cells to determine biomass in an environment of interest. Stains may be used to define and examine bulk tissues (e.g. muscle fibers or connective tissues), cell populations (different blood cells) or organelles within individual cells. Biological staining is also used to mark cells in flow cytometry, flag proteins or nucleic acids on gel electrophoresis Staining is not limited to biological materials, it can also be used to study the morphology (form) of other materials e.g.
    [Show full text]
  • Permanent Slides of Plant Cuticle Stained with Sudan IV and Sudan Black B
    Proceedings of the Iowa Academy of Science Volume 49 Annual Issue Article 14 1942 Permanent Slides of Plant Cuticle Stained with Sudan IV and Sudan Black B H. L. Dean State University of Iowa Edwards Sybil Jr. State University of Iowa Let us know how access to this document benefits ouy Copyright ©1942 Iowa Academy of Science, Inc. Follow this and additional works at: https://scholarworks.uni.edu/pias Recommended Citation Dean, H. L. and Sybil, Edwards Jr. (1942) "Permanent Slides of Plant Cuticle Stained with Sudan IV and Sudan Black B," Proceedings of the Iowa Academy of Science, 49(1), 129-132. Available at: https://scholarworks.uni.edu/pias/vol49/iss1/14 This Research is brought to you for free and open access by the Iowa Academy of Science at UNI ScholarWorks. It has been accepted for inclusion in Proceedings of the Iowa Academy of Science by an authorized editor of UNI ScholarWorks. For more information, please contact [email protected]. Dean and Sybil: Permanent Slides of Plant Cuticle Stained with Sudan IV and Sudan PERMANENT SLIDES OF PLANT CUTICLE STAINED WITH SUDAN IV AND SUDAN BLACK B H. L. DEAN AND EDWARD SvmL, JR. Sudan IV is commonly used to stain fats, oils, suberin, and cut­ in. Materials stained in this dye are usually mounted temporarily in glycerine and are seldom kept as permanent slides. This may be due to the fact that balsam, clarite or similar mounting media, cannot be used to make permanent slides of preparations stained in Sudan IV. The dye is immediately removed by the xylene or toulene solvent of these media, leaving the preparations colorless.
    [Show full text]
  • Safranin-O and Type II Collagen Stains WEB-PR-PT-2501 SAF-2
    Lonza Walkersville, Inc. www.lonza.com [email protected] Tech Service: 800-521-0390 Document #WEB-PR-PT-2501 SAF-2 02/08 Walkersville, MD 21793-0127 USA © 2008 Lonza Walkersville, Inc. Safranin-O and Type II Collagen Stains for In Vitro Chondrogenesis Materials • 10% Formalin, neutral buffered – VWR catalog #VW3239-4 or equivalent • 100% Ethanol • Xylene or xylene substitute • Eosin stain • Paraffin • Weigert’s iron hematoxylin • Fast Green FCF • 1% acetic acid • 0.1% aqueous Safranin-O • Proteinase K – DAKO code # S3020 or equivalent • Immunohistochemical staining kit – DAKO code # K4006 or equivalent • Type II Collagen specific primary antibody • Harris hematoxylin • Chondrogenic pellet cultures Procedure Chondrogenic Pellet Processing • Fix each chondrogenic cell pellet in 10% neutral buffered formalin for 1-24 hours at room temperature • Transfer each pellet to the corner of a small histology transfer bag, fold the bag and place it into a small histology cassette, labeled in pencil • Transfer the cassette to 70% ethanol. The pellets remain in the cassettes throughout processing until embedded • Dehydrate pellets in successive ethanol washes of 70%, 70%, 80%, 80%, 95% and 95%, for 15 minutes each • Stain pellets with Eosin by dipping briefly in Eosin, followed by three brief rinses in 100% ethanol. This stain serves only to visualize the pellets and aid in their handling. The pellets will stain orange • Incubate pellets in two changes of 100% ethanol, 20 minutes each • Transfer to xylene or xylene substitute, two changes, 20 minutes
    [Show full text]
  • Safranine O Gurr® for Microscopical Staining
    In Vitro Diagnostic Medical Device For professional use only Safranine O Gurr® for microscopical staining Cat. No Pack Type Pack Size Using an ignited loop, transfer a quantity of specimen, which 343122N Glass Bottle 25 g may be a body fluid, exudate, pus, or a liquid or solid culture, on to a degreased slide. Then distribute the specimen either Composition directly or after adding 1-2 drops of physiological saline C.I. No. 50240 >80% solution. After drying in air, heat-fix the smear C H ClN Heat –fixation: 20 19 4 Dry the smear at room temperature.Grip the slide and pass M = 350.85 g/mol through the flame of a Bunsen Burner several times to heat- kill and adhere the organism to the slide Intended Use(s) Leave to cool and stain. Dye for Gram stain procedures .Gram staining is a method All samples must be clearly labelled. used for direct microscopical examination to differentiate bacterial species into two large groups (Gram-positive and Gram-negative) based on the physical properties of their cell Preparation walls Safranin O stock solution The Gram stain provides prelimary identification information Dissolve 2.5 g safranin O Gurr® in 100 ml of 96% ethanol. and is not a substitute for culture studies for the diagnosis of For use, 10 ml of solution should be diluted with 90 ml of infectious diseases in human distilled water. Futher tests must be selected and implemented acc. to recognized methods for identification Staining on a staining bench Samples are derived from the human body, and heat fixed on 1.
    [Show full text]
  • THE SUITABILITY of CERTAIN STAINS for STUDYING LIGNIFICATION in BALSAM FIR, ABIES BALSAMEA (L.) MILL.1 Norman P
    The University of Maine DigitalCommons@UMaine Technical Bulletins Maine Agricultural and Forest Experiment Station 3-1-1972 TB53: The uitS ability of Certain Stains for Studying Lignification in Balsam Fir, Abies balsamea (L.) Mill Norman Kutscha James R. Gray Follow this and additional works at: https://digitalcommons.library.umaine.edu/aes_techbulletin Part of the Wood Science and Pulp, Paper Technology Commons Recommended Citation Kutscha, N.P., and J.R. Gray. 1972. The uits ability of certain stains for studying lignification in balsam fir, Abies balsamea (L.) Mill. Life Sciences and Agriculture Experiment Station Technical Bulletin 53. This Article is brought to you for free and open access by DigitalCommons@UMaine. It has been accepted for inclusion in Technical Bulletins by an authorized administrator of DigitalCommons@UMaine. For more information, please contact [email protected]. The suitability of certain stains 3r studying lignification in balsam fir, ibies balsamea (L.) Mill Kutscha and Gray Technical bulletin 53 March 1972 Life Sciences and Agriculture Experiment Station Cover photo: Safranin and aniline blue, showing blue unlignified cambium (top of photo) and contrasting red lignified tissue (lower part of photo). Imma­ ture, secondary walls appear blue-blue green and can be seen approximately five to eight cells down from the cambial region. Section of FAA-killed and celloidm embedded compression wood sample collected July 6, 1966; X 320. ABSTRACT An investigation was conducted to examine the suitability of ten staining reactions for studying lignification in balsam fir, Abies balsamea (L.) Mill. Two experiments were carried out on material collected on two different dates.
    [Show full text]
  • The Effect of Safranin on Glutathione Reductase and Glucose 6
    M. Kuzu et al. / Hacettepe J. Biol. & Chem., 2011, 39 (2), 189–194 The Effect of Safranin on Glutathione Reductase and Glucose 6-Phosphate Dehydrogenase Enzymes Glutatyon Redüktaz ve Glukoz 6-Fosfat Dehidrogenaz Enzimleri Üzerine Safraninin Etkisi Research Article / Araştırma Makalesi Müslüm Kuzu1,2, Murat Şentürk1,2* , Mehmet Çiftci2 1Ağrı İbrahim Çeçen University, Faculty of Art and Science, Department of Chemistry, Ağrı, Turkey 2Atatürk University, Faculty of Science, Department of Chemistry, Erzurum, Turkey ABSTRACT he aim of this study was to assess inhibitory effect of safranin on human erythrocyte glutathione reductase T(GR) and glucose 6-phosphate dehydrogenase (G6PD). For this purpose, human erythrocyte glutathione reductase was purified 2175-fold (31.35% yield) using 2’,5’-ADP Sepharose 4B affinity gel and Sephadex G-200 gel filtration chromatography and erythrocyte glucose 6-phosphate dehydrogenase was purified 8681.3-fold (24.77% yield) using 2’,5’-ADP Sepharose 4B affinity gel. in vitro Safranin exhibited inhibitory effects on GR and G6PD enzymes . The IC50 values of safranin were ± ± 1.15 mM for GR and 3.22 mM for G6PD, respectively, and the Ki constants were 2.63 0.13 and 4.53 0.17 mM, respectively. Safranin displayed noncompetitive inhibition for GR and G6PD. Key Words Safranin; glutathione reductase; glucose 6-phosphate dehydrogenase. ÖZET u çalışmanın amacı safraninin insan eritrosit glutayon redüktaz (GR) ve glukoz 6-fosfat dehidrogenaz B(G6PD) üzerindeki inhibitör etkilerini değerlendirmektir. Bu amaçla, 2’,5’-ADP Sepharose 4B afinite jeli ve Sephadex G-200 jel filtrasyon kromatografisi kullanılarak insan eritrosit glutayon redüktaz 2175-kat (%31.35 verim) ve 2’,5’-ADP Sepharose 4B afinite jeli kullanılarak eritrosit glukoz 6-fosfat dehidrogenaz 8681.3-kat (%24.77 verim) ile saflaştırıldı.
    [Show full text]