Biological Stain Commission Membership,May 1949

Officers

President, H. J. Conn, Box 269, Geneva, N. Y. (representing Society of American Bacteriologists.)

Vice-President, W. F. Windle, University of Pennsylvania, Philadelphia, Pa. Secretary, S. I. Kornhauser, University of Louisville Medical School, Louisville

Ky. Treasurer, E. H. Stotz, University of Rochester, School of Medicine, Rochester

N. Y. (representing American Chemical Society.)     

Book Review

LEGGETT, W. F. Ancient and Medieval Dyes. 5 × 8 in. 96 pp. Cloth. Chemical Publishing Co., Brooklyn, N. Y. 1944. $2.25.

Microtechnic In General. McCARTNEY, J. E. A new immersion oil “polyric”. J. Path. & Bact., 56, 265-6. 1944.

NICKERSON, MARK. A dry ice freezing unit for rotary microtomes. Science, 100, 177-8. 1944.

Dyes And Thedx Biological Uses. BERGEIM, FRANK H., and BRAKER, WILLIAM. Homosulfanilamides. J. Amer. Chem. Soc., 66, 1459. 1944.

CALDWELL, W. T., TYSON, F. T., and LAUER, LOTHAR. Substituted 2-sulfonamido-5-aminopyridines. II. J. Amer. Chem. Soc., 66, 1479. 1944.

JOHNS, C. K. Dye concentration in resazurin tablets. Amer. J. Pub. Health, 34, 955-8. 1944.

SMITH, WINSLOW WHITNEY. Relative sensitivity of different phases of growth curve of Bact. salmonicida to alkaline acriflavine. Proc. Soc. Exp. Biol. & Med., 56, 240-2. 1844.

VAN ARENDONK, A. M., and SHOULE, H. A. Dialkylaminoalkyl derivatives of substituted quinolines and quinaldines. J. Amer. Chem. Soc., 66, 1284. 1944.

WHEELER, KEITH, and DEGERING, E. F. Preparation and properties of certain derivatives of sulfamide. J. Amer. Chem. Soc., 66, 1242. 1944.

Animal Microtechnic. BOARDMAN, EDWARD T. Methods for collecting ticks for study and delineation. J. Parasitology, 30, 57-9. 1944.

DICKIE, MARGARET M. A new differential stain for mouse pituitary. Science, 100, 297-8. 1944.

GOVAN, A. D. TELFORD. Fat staining by Sudan dyes suspended in watery media. J. Path. & Bact., 56, 262-4. 1944.

LILLIE, R. D., and ASHBURN, L. L. Supersaturated solutions of fat stains in dilute isopropanol for demonstration of acute fatty degenerations not shown by Herxheimer technic. Arch. Path., 36, 432. 1943.

MULLEN, J. P. A convenient and rapid method for staining glycogen in paraffin sections with Best's carmine stain. Amer. J. Clin. Path., Tech. Sect., 8, 9-10. 1944.

NYKA, W. A method for staining the rickettsiae of typhns in histological sections. J. Path. & Bact., 56, 264. 1944.

POPPER, HANS, GYORGY, PAUL, and GOLDBLATT, H. Fluorescent material (ceroid) in experimental nutritional cirrhosis. Arch. Path., 37, 161. 1944.

SMALL, C. S., and SCHULTZ, M. A. Sustaining faded tissue sections. Amer. J. Clin. Path., Tech. Sect., 7, 66-7. 1943.

YOFFEY, J. M., and PARNELL, J. The lymphocyte content of rabbit bone marrow. J. Anat., 78, 109-12. 1944.

ZIEGLER, E. E. Hematoxylin-eosin tissue stain. An improved, rapid, and uniform technic. Arch. Path., 37, 68. 1044.

Plant Microtechnic. HAASIS, FERDINAND W. Staining rubber in ground or milled plant tissues. Ind. and Eng. Chem., Anal. Ed., 16, 480. 1944.

PARRIS, G. K. A simple nuclear stain and staining technique for Helminthosporia. Phytopathology, 34, 700. 1944.

Microorganisms. DARZINS, E. Rickettsienstudien. Zentbl. Bakl., Abt. I, Orig., 151, 18-20. 1943.

GOHAR, M. A. A staining method for Corynebacterium diphtheriae. J. Bact., 47, 575. 1944.

GRAY, P. H. H. Two-stain method for direct bacteria count. J. Milk Techn., 6, 76. 1943.     

Book Reviews

Free Radicals in the Brain-Aging, Neurological and Mental Disorders L. Packer, L. Prilipko and Y. Christen (Eds.)

Ginkgo biloba Extract (EGb 761) as a Free Radical Scavenger, Advances in Ginkgo biloba extract research, volume 2 Edited by C. Ferradini, M.T. Droy-Lefaix and Y. Christen, Elsevier, Paris, 1993, 186 pages     

Book Reviews

Oxygen, Gene expression and Cellular Function, Vol 105 Lung Biology in Health and Disease L. B. Clerch and D. J. Massaro Marcel Dekker, 1997

Food and Free Radicals Edited by Midori Hiramatsu, Toshikazu Yoshikawa and Masayosu Inoue Plenum Press, 1997, ISBN 0-306-45493-9 vii + 169 pages

Preventing Coronary Heart Disease: The Role of Antioxidants, Vegetables and Fruit Ed Lesley Rogers and Imogen Sharp National Heart Forum 1997 The Stationery Office London

Inducible Gene Expression Volume 1 Environmental Stresses and Nutrients Volume 2 Hormonal Signals Ed by PA Baeuerle, Birkhauser Verlag AG, Basel, 1997

Antioxidants in Science, Technology, Medicine and Nutrition Gerald Scott Albion Chemical Science Series, Ellis Horwoood Publishing Ltd, Chichester, UK, 1997

Flavonoids in Health and Diseases Eds C A Rce-Evans and L Packer Marcel Dekker Inc, New York, 1997     

Book Reviews

The Biology of Nitric Oxide, vol. 3. Physiological and Clinical Aspects; vol. 4. Enzymology, Biochemistry and Immunology, Portland Press, 1994

Mitochondrial Diseases Editors: L. Ernster, R. Luft and S. Orrenius Publishers: Elsevier

Biothiols in Health and Disease Editors: Lester Packer & Enrique Cadenas Publishers: Marcel Dekker, Inc.     

Laboratory Hints from the Literature

Microscope and Other Apparatus: Bernard, J. E. The principles of fluorescence microscopy. J. Royal Micro. Soc., 57, 256-9. 1937.

Wrighton, H. A new light source for microscopy. J. Royal Micro. Soc., 57, 260-1. 1938.

Microtechhic in general: Comandon, J. and De Fonbrune, P. La chambre $aG huile. Ses avantages pour l'$eGtude des microorganisms vivants, la culture des tissus et la micromanipulation. Ann. Inst. Pasteur, 60, 113-41. 1938.     

Book Review

Salmon, M. V. Practical phase-contrast microscopy. The Microscope, 6, 177-88. 1947.

Evans, Titus C. Radioautographs in which the tissue is mounted directly on the photographic plate. Proc. Soc. Exp. Biol, and Med, 64, 813-15. 1947.

Tolbert, B. M., and Branch, G. E. K. The spectra of the doubly charged positive ions of some p,p'-diaminotriphenylmethane dyes. J. Amer. Chem. Soc, 69, 1083-81. 1947.

Van Wyk, J. J., and Clark, W. M. The luminosity and chromaticity of indicators as a function of pH. J. Amer. Chern. Soc, 69, 1296-1301. 1947.

Hamre, Christopher J. Hematopoiesis in the bone marrow of rats recovering from nutritional anemia. J. Lab. & Clin. Med, 32, 756-76. 1947.

Limarzi, Louis R. Evaluation of bone marrow concentration techniques. A modified method for the simultaneous preparation and staining of blood and bone marrow films. J. lab. & Clin. Med, 32, 782-40. 1947.

Heath, O. V. S. Role of starch in light-induced stomatal movement, and a new reagent for staining stomatal starch. Nature, 159, 647-8. 1947.

Cohen, H. A. A new quick method for staining Treponema pallidum. Acta Med. Orientalia, 6, 99-100. 1947.

Henry, H., and Stacey, M. Histochemistry of the Gram-staining reaction for micro-organisms. Proc. Roy. Soc, Ser. B. Biol. Sci, 133, 891-406. 1946.

Para, M. Silver impregnation of spirochetes in tissue sections. Arch. Path, 42, 649. 1946.

Ruiz, Merino, J. A method of staining capsules. Rev. Sanidad e Hig. Publ (Madrid), 20, 1112. 1946.     

The Fixing and Staining of Liriodendron Tulepifera Root Tips and their Mycorrehizal Fungus

Root tips of Liriodendron Tulipifera forming with a fungus of the Mycelium Radicis group an endotrophic mycorrhiza, were subjected to several different fixations in which the action of cationic chromium and anionic chromium were compared. Anionic chromium in the form of chromic acid was combined with several substituted benzene compounds while cationic chromium in the form of chromic sulfate (Cr₂(SO₄)₃·15 H₂O) in 4% formaldehyde (HCHO) was used with the same ring compounds. In addition, several fixatives not containing chromium were tested.

Five percent chromic sulfate (Cr₂(SO₄)₂·15 H₂O) in 4% formaldehyde (HCHO) or in 1% osmic acid with saturated aqueous salicylic and/or picric acid preserved the histological and cytological details of the mycorrhiza, as was clearly demonstrated when followed by staining in 3% acetic acid saturated with orseillin BB and counterstained with 1% crystal violet in clove oil.

Two percent ferric chloride (Fe Cl₃) in 4% formaldehyde showed the relationship of the tannin contents of the cells to the invading hyphae when followed by suitable staining.

Cationic chromium appeared to be superior to anionic chromium in preserving cell walls as well as the general histologic features of the material investigated.     

Book Reviews

Mutation Research DNAging Genetic Instability and Aging

Cardiovascular Toxicology Second edition, edited by Daniel Acosta, Jr. Raven Press, New York, USA. $124, 1992

Dietary Fats: Determinants of Preference, Selection and Consumption Edited by D.J. Mela Elsevier Applied Science, 1992, ix + 192 pages Price $75.00 ISBN 1-85166-865-9

Determination of Vitamin E: Tocopherols and Tocotrienols by Claude Bourgeois Elsevier Applied Science, 1992, vi + 162 pages Price $75.00 ISBN 1-85166-7547

Biochemistry of Food Proteins Edited by B.J.F. Hudson Elsevier Applied Science, 1992, x + 419 pages Price $110.00 ISBN 1-85166-768-7

Lipid-soluble Antioxidants: Biochemistry and Clinical Applications Edited by A.S.H. Ong and L. Packer Birkhauser Verlag, 1992, xii + 640 pages Price SFR 168.00 (approx $76) ISBN 3-7643-2667-0 (Basel) ISBN 0-8176-2667-0 (Boston)     

Acid Versus Neutral Formalin Solution as a Neurological Fixative

The fixing action of 10% formalin solution alone and with formic, acetic, propionic, butyric, lactic, monochloracetic, dichloracetic, or trichloracetic acid was studied by means of stains with silver, osmic acid and cresyl violet. The following conclusions were reached:

1. In general, better fixation and staining was obtained with acid than without.

2. Less difference was seen in comparing one acid with another than was expected before the experiments were made.

3. Propionic, butyric, and dichloracetic showed no promise of having practical value.

4. Formic and monochloracetic acids as modifiers gave superior stains with osmic acid, while silver and cresyl violet stains of the same material were about equal to those made from formalin-acetic fixed material.

5. Lactic acid caused somewhat more distortion of tissue elements than the others but was compatible with good staining.

6. Acetic acid was most effective in concentrations of 3 to 5% while the stronger acids such as formic, monochloracetic, lactic and trichloracetic were effective in concentrations of 0.5 to 1%.     

Book Review

DNA and Free Radicals Halliwell B. and Aruoma O.I. (Eds). Ellis Harwood: London, 1993, p. 332 ISBN 0132220350.

Lipids: The Continuing Challenge American Journal of Clinical Nutrition May 1993 Vol 57 NO 5(s)     

Laboratory Hints from the Literature

DYES AND THEIR BIOLOGICAL USES Burckhalter, J. H., Jones, E. M., Holcomb, W. F., and Sweet, L. A. N-substituted 2-methoxy-6-chloro-9-aminoacridines. J. Amer. Chem. Soc., 65, 2012. 1943.

Galat, Alexander. New processes for sulfanilamide. Ind. and Eng. Chem., Ind. Ed., 36, 192. 1944.

Kumler, W. D., and Daniels, T. C. The relation between chemical structure and bacteriostatic activity of sulfanilamide type compounds. J. Amer. Chem. Soc., 65, 2190. 1943.

Kwartler, C. E., and Lucas, Philip. The preparation of sutfanil-amidoindazoles. J. Amer. Chem. Soc., 65, 1804. 1943.

Mueller, A. C., and Hamilton, C. S. The synthesis of 1-substituted aminobenzo(f)quinolines. J. Amer. Chem. Soc., 65, 1017. 1943.

Popkin, A. H. Derivatives of biphenylsulfonamides. I. Preparation of p-(o-aminopnenyl)-benzenesulfonamide. J. Amer. Chem. Soc., 65, 2043. 1943.

Popkin, A. H., and Perretta, Gertrude M. Derivatives of biphenyl-sulfonamide. II. Derivatives of p-(o-aminophenyl)-benzenesulfonamide. J. Amer. Chem. Soc., 65, 2046. 1943.

Shreve, R. N., and Bennett, R. B. Studies in azo dyes. I. Preparation and bacteriostatic properties of azo derivatives of 2,6-diaminopvridine. J. Amer. Chem. Soc., 65, 2241. 1943.

Shreve, R. N., and Bennett, R. B. Studies in azo dyes. II. Preparation and bacteriostatic properties of azo derivatives of 8-quinolino. J. Amer. Chem. Soc., 65, 2243. 1943.

Siebenmann, C., and Schnitzer, R. J. Chemotherapeutic study of p-nitrobenzoyl- and related compounds. J. Amer. Chem. Soc., 65, 2127 1943.

ANIMAL MICROTECHNIC Barrett, A. M. A method for staining sections of bone marrow. J. Path & Bact., 56, 133-5. 1944.

Barrett, A. M. On the removal of formaldehyde-produced precipitate from sections. J. Path. & Bact., 56, 135-6. 1944.

Chang, Min-Chueh. Disintegration of epidymal spermatozoa by application of ice to the scrotal testis. J. Exp. Biol., 20, 16-22. 1943.

Ercoli, N., and Lewis, M. N. The age factor in response of bone tissue to alizarin dyes and the mechanism of dye fixation. Anat. Rec., 87, 67-76. 1943.

Hess, Manfred, and Hollander, Franklin. Permanent metachromatic staining of gastric mucus smears. J. Lab. and Clin. Med., 29, 321-3. 1944.

Miller, John A. A new method of staining nervous tissue. Ohio J. of Sci., 44, 31-5. 1944.     

Book Reviews

Gabe, Manfred. Histoiogical Techniques. (Translated by Robert Blackith and Aries Kovoor.) 1106 pages, 16 × 24 cm. 2 figures, 35 tables, hard covers, cloth bound. Masson, Paris and New York (and Springer, Heidelberg and New York), 1976. $49.50.

Goldman, R., Pollard, T. and Rosenbaum, J., Editors. Cell Motility. 3 volumes, 1373 pages, 18 × 26 cm. Proceedings of 3rd Cold Harbor Conference on Cell Proliferation. Cold Spring Harbor Laboratory, 1976.

Lillie, R. D. and Fullmer, H. M. Histopathologic Technic and Practical Histochemistry. Fourth Edition. 942 pages, 16 × 23 cm. 28 figures, 126 tables. McGraw-Hill, New York, 1976. $40.00.     

Marchi's Staining Method: Studies of Some of the Underlying Mechanisms Involved

Spinal cord of cat and rabbit was stained, after experimental lesions, by variations of Marchi's method. The following conclusions were drawn:

1. The presence of an oxidizing agent (K₂Cr₂O₇, NaIO₃, or KCIO₃) in the osmic acid solution is of primary importance and a preliminary oxidation in Mueller's fluid is unnecessary or even detrimental.

2. Acetic acid added to Marchi's fluid, accentuates the action of the oxidizing agent in restraining the staining of normal myelin.

3. Too high concentration of oxidizing agent or of acid may inhibit staining of degenerating myelin.

4. Marchi's and Busch's methods have been modified as follows: Fix one day in 10% formalin and transfer without washing to the staining mixture, either A or B. Staining mixture A: Marchi's fluid plus 1 to 3% glacial acetic acid. B: An aqueous solution containing KCIO₃ 0.25%, osmic acid 0.33%, and acetic acid 1%. Stain about one week. These methods worked on spinal cord and medulla, but cannot be recommended for brain.

5. The detrimental effects of long post mortem autolysis or of prolonged fixation in formalin may be counteracted to some degree by increasing the concentration of the acid in Marchi's fluid up to 5% or of the KCIO₃ up to 0.4% in the modified Busch's fluid.     

Marchi's Staining Method: II. Fixation

Following experimental lesions, spinal cords of cats and rabbits were fixed with acid, neutral, and alkaline solutions. Staining was limited to a chromate-osmic (Marchi's) solution and a chlorate-osmic solution. The following conclusions were drawn:

The presence of an acid in the fixative caused normal myelin sheaths to stain. This effect was reduced by washing tissues before staining, by adding acetic acid to the stain, or by employing a non-formalin fixative. It was, however, at no time entirely obviated.

A study was made of the granular deposits which occur in nearly all Marchi preparations and which are especially confusing if very light backgrounds are obtained.

The staining reactions of the granular deposits were very similar to those of degenerating myelin but some suppression of the granules was obtained by adding KCIO₃ to the formalin fixative.     

An Improvement in Staining Technic for Protozoa

A modification of Donaldson's iodine-eosin stain for staining intestinal protozoa is presented. This modification consists of using high dilutions of colloidal iodine (Chandler)² instead of Lugol's solution as well as high dilutions of eosin. A better resolution of the external and internal structures is brought about by the new method.

The procedure is as follows: A portion of the fecal material to be examined is suspended in a 0.6% salt solution; the suspension should be of a consistency so that one drop will make a satisfactory microscope mount under a cover glass. To ten parts of this suspension, in a test tube, is added one part of the stain which is prepared as follows:—

10 parts of distilled water

6 parts of a suspension of colloidal iodine (Chandler) containing 4% iodine—20% iodine suspensoid, Merck

1 part of a 10% water solution of anilin red, Merck (eosin yellowish)

Technicians will find, because iodine in the form of colloidal iodine is readily released to the organisms, that the use of this material is far superior to Lugol's solution hi carrying out the technic for staining intestinal protozoa in the study of fresh mount preparations. Not only are organisms more deeply stained with iodine but by eosin as well, even when employed in high dilutions.     

Book Reviews

Analysis of Free Radicals in Biological Systems Edited by A. Favier, J. Cadet, B. Kalyanaraman, M. Fontecave and J.-L. Pierre Birkhauser Verlag AG, Basel, 1995. DM 178

Essays in Biochemistry Volume 291995 Edited by DK Apps and KF Tipton     

Micro-Manipulation of Cells in Mammals

An arrangement of apparatus for applying micro-manipulative procedures to cells in living mammals is described. It was found satisfactory for manipulation of pericapillary cells and capillary endothelium in the greater omentum of the cat.

An animal board, a Bausch and Lomb triple purpose micro-projector and a double Fitz micro-manipulator were mounted on a spring platform. The micro-needles were drawn from Pyrex rods by hand. The stage of the microscope was modified to protect the condenser from fluids. Warm saline solution was carried to the exposed omentum thru flexible rubber tubing.

The use of a micro-dissection chamber which immobilized the part of the omentum under manipulation is explained. The construction of this chamber is shown by diagrams. A camera support was bolted to the base of the micro-manipulator.

The arrangement of apparatus is shown by a photograph.     

Preliminary Notes on Chromic Fixation in Alcoholic Media

In Note I are pointed out certain chemical difficulties which stand in the way of experiments on chromium fixation in alcoholic media; also a procedure is described by means of which these difficulties may be avoided in the preparation of a chrome-alcohol stock solution.

In Note II it is suggested that the properties of OsO₄ most important in the quick-killing effect conferred by this adjuvant upon aqueous chromic reagents are probably, in addition to its high toxicity, its volatility and its oxidant action. Volatile, toxic oxidants which do not attack alcohol are therefore discussed briefly, with reference especially to their use as quick-killing adjuvants in chrome-alcohol reagents. Iodine is noted as probably the most important inorganic possibility. Reasons are stated for considering the quinones the most promising of numerous possible organic compounds.

Results of fixations of Vicia faba root tips in nine variations of the proportions of acetic acid and iodine in a chrome-alcohol-aceticiodine combination are described. The chrome-alcohol-iodine reagents which proved best proportioned for V. faba root tips preserved some details of the chromosomes better than does Bouin's solution, but did not make certain details of the early prophase as clear as does Benda's modification of the Flemming reagent, according to a comparison with Sharp's figures of Bouin and Benda preparations of root tips of the same species.