Showing papers in "Biotechnic & Histochemistry in 1974"
TL;DR: The procedure is a simple, rapid staining technique suitable for photomicrography and tissue orientation for electron microscopy, and considerable variation was found in methylene blue-azure II staining times for different embedding media.
Abstract: One micron-thick sections of tissues fixed in glutaraldehyde, or in glutaraldehyde followed by osmium tetroxide, and embedded in a variety of plastic resins were stained in a methylene blue-azure II solution at 65 C, then counterstained in 0.05% basic fuchsin in 2.5% ethanol at room temperature (24 C). Considerable variation was found in methylene blue-azure II staining times for different embedding media. Aged Epon-812 required less staining time than freshly polymerized blocks of Epon-812. The procedure is a simple, rapid staining technique suitable for photomicrography and tissue orientation for electron microscopy.
499 citations
TL;DR: It is found that thionin can be reliably used as a polychrome stain for sections of neural tissue obtained from a freezing microtome.
Abstract: The need for rapid histological feedback on neural tissue is ever present. Although there are several stains which can be readily used for staining either cell bodies or fiber tracts, adequate contrasting stains which are both rapid and easy to apply are not generally available. In 1936 Chang presented a technique for whole brains utilizing the metachromatic properties of thionin. Unfortunately this procedure was very time consuming. For the last several years we have worked with several variations of this stain and have found that thionin can be reliably used as a polychrome stain for sections of neural tissue obtained from a freezing microtome.
161 citations
TL;DR: Fresh, ground, mineralized bone sections 75-100 μ thick are stained 90 minutes or 48 hours in the Bone Stain, a preparation containing fast green FCF, orange G, basic fuchsin, and azure II that helps in the differential diagnosis of certain metabolic bone diseases in huma...
Abstract: Fresh, ground, mineralized bone sections 75-100 μ thick are stained 90 minutes or 48 hours in the Bone Stain, a preparation containing fast green FCF, orange G, basic fuchsin, and azure II. Surface stain is then removed by grinding under running water. Sections are washed in 0.1% zephiran chloride (benzalkonium chloride) or in 0.01% mild soap and again washed in tap water, followed with distilled water. Sections are next differentiated in 0.01% acetic acid in 95% methanol, dehydrated in 95% ethanol and 100% ethanol, cleared in alcohol:xylene 1:1, 1:4, 1:9 and 2 changes of xylol, and then mounted permanently in Eukitt's mounting media.Osteoid seams stain either green to jade green or red to dark red, incompletely mineralized bone red or orange yellow, and the zone of demarcation light green. The walls of lacunae, canaliculae, feathered bone, procedural artifacts and periosteocyte lacunar low-density versions stain red.The method helps in the differential diagnosis of certain metabolic bone diseases in huma...
139 citations
TL;DR: Modifications of the neutral red vital staining technique of Drgsel and co-workers to sort live and dead marine copepods have made possible live-dead determinations of most components of estuarine zooplankton.
Abstract: Modifications of the neutral red vital staining technique of Drgsel and co-workers to sort live and dead marine copepods have made possible live-dead determinations of most components of estuarine ...
122 citations
TL;DR: Squash preparations of styles stained in watersoluble aniline blue and viewed under ultra-violet illumination are regularly used for examining pollen tubes because the callose plugs fluoresce brightly under these conditions, permitting easier, faster and more accurate assessments of pollen tube length and numbers in plum and pear styles.
Abstract: Squash preparations of styles stained in watersoluble aniline blue and viewed under ultra-violet illumination are regularly used for examining pollen tubes because the callose plugs fluoresce brightly under these conditions. Tubes are therefore clearly distinguish from the astylar tissue and may be readily counted and measured. This method has proved to be quite unsatisfactory for plum pollen tubes, since they contain very few cause plugs and better results have been obtained with a mixed stain of 0.1% aniline blue and 0.07% of the fluorescent brightener ‘Calcofluor White M2R New’. Styles are softened by autoclaving in 50 g/1 sodium sulphite, rinsed and stained for ten minutes, then squashed and examined with a fluorescence microscope in the usual way. Callose deposits, when present, fluoresce bright yellow, but lengths of tube with no deposits can also be clearly identified and followed, permitting easier, faster and more accurate assessments of pollen tube length and numbers in plum and pear styles.
94 citations
TL;DR: A procedure for the infiltration, polymerization, and sectioning of animal tissues in GMA for light microscopy is presented which is no more complex than paraffin techniques and which has a number of advantages.
Abstract: Glycol methacrylate (GMA), a water and ethanol miscible plastic, was introduced to histology as an embedding medium for electron microscopy. This medium may be made soft enough for cutting thick sections for routine light microscopy by altering its composition. A procedure for the infiltration, polymerization, and sectioning of animal tissues in GMA for light microscopy is presented which is no more complex than paraffin techniques and which has a number of advantages: (I) The GMA medium is compatible with both aqueous fixatives (formaldehyde, glutaraldehyde, Bouin's, and Zenker's) and non-aqueous fixatixes (Carnoy's, Newcomer's, ethanol, and acetone). (2) Undue solvent extraction of the tissue is avoided because adequate dehydration occurs during infiltration of the embedding medium. Separate dehydration and clearing of the tissue prior to embedding is eliminated. (3) When polymerized, the supporting matrix is firm enough that hard and soft tissues adjacent to one another may be sectioned without distort...
74 citations
TL;DR: The kitchen appliance known as the “radar oven” generates heat quickly in materials containing water, and protoplasm exposed to the irradiation can thus be denatured, and histological examination was carried out.
Abstract: The kitchen appliance known as the “radar oven” generates heat quickly in materials containing water. Protoplasm exposed to the irradiation can thus be denatured. The amount of heat generated is a function of the time of exposure and the intensity of the irradiation, and the size and specific heat of the tissue or organism being irradiated. But docs such heating have applicabiity to histological technique? One of four carcas temperaturea (approximately 60°, 70°, 77°, and 85 C) was generated in anaesthetized, adult hairless mice of both sexes. “Control” animals were not irradiated. Specimens of liver, kidney, lung, and (from males) testis were taken from the five groups; the tissue spedmens were dehydrated in tetrahydrofuran, embedded in paraffin, sectioned at 9 μm, and stained with hematoxylin and eosin. The preparations were suitable for histological examination. Each organ had an optimum temperature for histological fixation under the conditions of this experiment: liver, ∼70°; kidney, ∼77°; lung, ∼77°;...
64 citations
TL;DR: Rapid, onestep polychromatic staining of 0.75-1.5 μm epoxy sections of glutaraldehyde-osmium fixed tissues can be obtained with mixtures of basic fucbsin and toluidme blue O in alkaline polyethylene glycol ZOO (PEG ZOO).
Abstract: Rapid, onestep polychromatic staining of 0.75-1.5 μm epoxy sections of glutaraldehyde-osmium fixed tissues can be obtained with mixtures of basic fucbsin and toluidme blue O in alkaline polyethylene glycol ZOO (PEG ZOO). Sections are attached to slides by heating at 100 C for 45 seconds and stained at that temperature for 2-3 minutes with a solution consisting of PEG 200 (50 ml), 0.2 N KOH (0.75 ml), basic fuchsin (1.7 gm), and toluidine blue O (0.3 gm). Red-blue balance and selective staining of different structures can be controlled by varying the amount of toluidine blue added. After rinsing with 10% acetone and rapid drying, sections are covered with immersion oil or mounting medium and a cover-slip. Total time from cutting of a section to finished preparation is less than 6 minutes. This staining solution is stable, does not produce precipitates on the sections, and does not wrinkle or lift the sections from the slides.
59 citations
TL;DR: Haupt's Gelatin Adhesive Mixed with Formalin for Affixing Paraffin Sections to Slides as discussed by the authors was the first gelatin adhesive mixed with formalin for affixing paraffin sections to slides.
Abstract: (1974). Haupt's Gelatin Adhesive Mixed with Formalin for Affixing Paraffin Sections to Slides. Stain Technology: Vol. 49, No. 2, pp. 116-117.
52 citations
TL;DR: A critical point drying method using dry ice (solid carbon dioxide) instead of liquid carbon dioxide is reported, which needs no gas cylinder and even minute or fragile specimens do not blow away because there is no flow of gas.
Abstract: A critical point drying method using dry ice (solid carbon dioxide) instead of liquid carbon dioxide is reported. After the specimens are placed in the chamber of the medical point drying apparatus, dry ice cut to the shape of the chamber is inserted. The chamber is closed and warmed to change the dry ice into liquid carbon dioxide. This method needs no gas cylinder and even minute or fragile specimens do not blow away because there is no flow of gas. This method can be used with any kind of critical point drying apparatus.
41 citations
TL;DR: A convenient, rapid and extremely effective thin-layer chromatographic system for the analysis of Romanowsky-type blood stains is reported.
Abstract: A convenient, rapid and extremely effective thin-layer chromatographic system for the analysis of Romanowsky-type blood stains is reported. The system employs silica ad absorbent, and as developing solvent, the butanol layer obtained from butan-1-al (12 vol), 1% w/v aqueous ammonium chloride (5 vol) and 2% v/v aqueous formic acid (2 vol).The separations obtained are illustrated using a commercial sample of each of the following stains: azure A (CI 52005), azure B (CI 52010), azure C (CI 52002), methylene blue (CI 52015), methylene violet Fkrnthsen, eosin bluish (CI 454500), eosin Y (CI 45380), Giemsa stain, Jenner stain, Leishman stain, MayCrunwald stain, Romanowsky stain, Wright stain.
TL;DR: “Dirt” on electron microscopic sections can generally be avoided by the simple strategem of preventing dry sections from coming in contract with any solution with a dirty surface layer.
Abstract: “Dirt” on electron microscopic sections can generally be avoided by the simple strategem of preventing dry sections from coming in contract with any solution with a dirty surface layer. Wet sections can be pushed through the surface layer of such solutions without ill effect. The first and last solutions to touch a section must be dean, preferably distilled water from a plastic wash bottle.Mention of a trademark name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA, nor does it imply its approval to the exclurion of other products that may also be suitable.
TL;DR: An improved procedure for quantitation of cotton fiber development, the “stain-destain” method, is reported, which is deemed accurate and precise for the purpose intended—quadtation of fiber development as modified by phytohormones or other treatments.
Abstract: An improved procedure for quantitation of cotton fiber development, the “stain-destain” method, is reported. Toluidine blue 0 was used to selectively stain fibers subsequently destained in an acid-alcohol solution. Absorbance of the dyecontaining destaining solution was used as a measure of fiber development, and expressed in terms of total fiber units (TFU), one OD unit at 624 nm having been assigned the value of one TFU. Optimum conditions for the procedure, including staining and destaining times and solution to ovule ratios were determined: (1) 20 ovules with associated fibers stained for 15 sec in 80 ml 0.018% toluidine blue O, (2) nonabsorbed dye removed by 60 sec wash, (3) ovuls destajned in 100 ml glacial acetic acid-ethanol-water (10:95:5), (4) absorbance determined after one hr destaining. The procedure is deemed accurate and precise for the purpose intended—quadtation of fiber development as modified by phytohormones or other treatments. Data are shown correlating TFU with fiber length through ...
TL;DR: A method which gives good quality 1-2 μm thick sections of undecaldfied cancellous and thin cortical bones for light miuoscopy is described and the same specimens can be used to prepare thick sections for microradiography.
Abstract: A method which gives good quality 1-2 μm thick sections of undecaldfied cancellous and thin cortical bones for light miuoscopy is described. Formalin fixed material is dehydrated in graded acetones and embedded in a modiEed formula of Spurr's low viscosity embedding medium. After a 16 hour polymerisation period at 60 C, sections are cut at 1-2 μm thickness on a Porter-Blum JB4A rotary microtome Using glass knives. Sections are attached to clean glass slides with heat, the resin degraded in bromine vapour and removed in acetone. This allows comparative ease of staining. The technique is rapid, does not interfere with tetracycline fluorescence and the same specimens can be used to prepare thick sections for microradiography.
TL;DR: Longitudinal cryostat sections of skeletal muscle picked up with glass slides onto which a film of ethylenediaminetetracetic acid (EDTA) has been dried after dipping in a 3% EDTA solution do not show the severe retraction artifacts obtained when plain glass slides are used.
Abstract: Longitudinal cryostat sections of skeletal muscle picked up with glass slides onto which a film of ethylenediaminetetracetic acid (EDTA) has been dried after dipping in a 3% EDTA solution do not show the severe retraction artifacts obtained when plain glass slides are used. Histochemical techniques work well on the longitudinal sections after the EDTA has been rinsed away and remaining traces saturated with calcium, Morphologic preservation is good. Regularly distributed histochemical reaction products are found within any one fiber.
TL;DR: Batch variations in commercial dyes employed for Romanowsky-type staining have been studied by thin-layer chromatography and it has been found that of these dyes only methylene blue and thionine are commercially available reasonably free of colored contaminants.
Abstract: Batch variations in commercial dyes employed for Romanowsky-type staining have been studied by thin-layer chromatography.The colored components of numemus batches of the thiazine dyes azure A, azure B, azure C, methylene blue, methylene violet hthsen, thionine and tduidine blue, and of the xanthene dyes emin B and eosin Y, have been compared. It has been found that of these dyes only methylene blue and thionine are commercially available reasonably free of colored contaminants. The azures, methylene violet Bernthsen and toluidine blue are complex mixtures of thiazine dyes of the thionine and thionoline series. Samples of eosin B are mixtures of the nominal dye with win Y, fluorescein and tribromofluorescein. Thoee of eosin Y are mixtures of this dye with fluorescein and tribromofluorescein.
TL;DR: Frozen sections of musde fixed in buffered formaldehyde are first incubated for localization of esterase using 5-bromoindoxyl acetate as substrate and stained by a urea-silver nitrate method for axons.
Abstract: Frozen sections of musde fixed in buffered formaldehyde (pH 7.3) are first incubated for localization of esterase using 5-bromoindoxyl acetate as substrate. The sections are then mounted on slides and stained by a urea-silver nitrate method for axons. Result: subneural apparatus—blue; axons—black; other tissue components in various shades of grey.
TL;DR: The opinion is expressed that this method, based on the use of column chromatography, is also applicable to the purification of other cationic dyes.
Abstract: A method is described for the purification of the dye azure B in quantities sufficient for biological staining experiments on a larger scale. The method is based on the use of column chromatography. Two columns are employed. In column A with silica gel as adsorbent the azure B fraction is isolated from a suitable substrate ('technical' azure B gained by a modification of Bernthsen's synthesis of methylene blue, or polychrome methylene blue) using an acetate-formate mixture as eluent. In column B, on an Amberlite polymeric adsorbent (XAD-2) the acetate-formate anions are exchanged in chloride. Regeneration of both columns is possible: KMnO4, Na2S2O4 and water are run through column A; 5% NaOH, methanol and water through column B. Purification of azure B on economic terms is thus attained. The opinion is expressed that this method is also applicable to the purification of other cationic dyes.
TL;DR: A Modified Alcian Blue-Periodic Acid Schiff Stain for Epoxy-embedded Atherosclerotic Arteries as discussed by the authors is a modified Alcian blue-periodic acid Schiff Stain.
Abstract: (1974). A Modified Alcian Blue-Periodic Acid Schiff Stain for Epoxy-embedded Atherosclerotic Arteries. Stain Technology: Vol. 49, No. 4, pp. 241-243.
TL;DR: Results indicate that tissues are well stained even after an 8-week exposure, aldehyde-fuchsin pduces no chemographic effect, and structures underneath the emulsion are easily identified.
Abstract: Four-week-old Holtzman rats were injected intraperitoneally with 20 μCi 125I. Six or eight weeks later, they were killed by intracardiac perfusion with glutaraldehyde; thyroid and adrenal glands were excised, postfixed in osmic acid, and embedded in Epon. Steps in the staining procedure of 0.5-1 μm thick sections are: oxidation in 0.3% potassium permanganate in 0.625% sulfuric acid, 2-5 min at 70 C; brief rinse; bleaching with 2.5% NaHSO3, 4-5 min; brief r-utse; let dry completely; aldehydefuchsm, 15-20 min at 50 C; 95% alcohol; rinse in absolute alcohol; let dry completely. seaions were coated with Kodak NTB2 emulsion and exposed for 3 to 8 weeks. Results indicate that (1) tissues are well stained even after an 8-week exposure, (2) aldehyde-fuchsin pduces no chemographic effect, and (3) structures underneath the emulsion are easily identified.
TL;DR: A method for staining reticulin in formalin fixed, paraffin embedded sections is described, using a silver solution prepared volumetrically by mixing solutions of defined concentrations of ammonium nitnte, sodium hydroxide and silver nitrate.
Abstract: A method for staining reticulin in formalin fixed, paraffin embedded sections is described. The steps include oxidation in permangauate, impregnation in a silver solution, reduction in formalin and toning in gold. The permanganate is acidified, enhancing its oxidizing potency and decreasing the stainiig of background elements. The silver solution is prepared volumetrically by mixing solutions of defined concentrations of ammonium nitnte, sodium hydroxide and silver nitrate, instead of the drop by drop titration with ammonium hydroxide usually required. In consequence, the silver solution and the staining procedure as a whole are readily reproducible. The contrast between the deeply stained reticulin and the lightly stained background is marked, and permits an assessment of the distribution of reticulin under low magnification.
TL;DR: Crystal deposits in human kidney and thyroid were used to assess the staining of oxahtes by selected methods for calcium, and the silver nitrate-rubeanic acid sequence was found to give the best visualisation of the crystals.
Abstract: Crystal deposits in human kidney and thyroid, identified as calcium oxalate by microincineration and Solubility tents, were used to assess the staining of oxahtes by selected methods for calcium. No method that stained the crystals was considered specific for calcium oxalate, but, after removal of possible phosphate and carbonate with 2 M acetic acid, the silver nitrate-rubeanic acid sequence was found to give the best visualisation of the crystals, and could be considered reasonably selective. Deposits in kidneys included a pronounced colloidal matrix composed chiefly of acid mucopolysaccharides. This matrix often showed a lamellar pattern and was well-demonstrated by alcian blue at pH 25 and by dialysed iron after removal of the crystals with 1 N hydrochloric acid. Such a matrix Could only be detected in trace amount in the thyroid deposits.
TL;DR: The dexribed technique facilitates oriented embedding of individual cells in various media for both light and electron microscopy.
Abstract: The dexribed technique facilitates oriented embedding of individual cells in various media for both light and electron microscopy. A fixed Specimen is embedded in a small cube of 2% agar at 40 C and subsequently sealed in the desired orientation to a strip of black paper which then serves as a tab for transferring the specimen during dehydrating and embedding procedures. The beveled ends of the strip indicate the exact location of the specimen in the cube. This technique can be employed for the embedding media used in both light and electron microscopy. It ah permits photomicrographs of the whole specimen to be made which can be compared with photomicrographs of individual sections cut from the specimen in a selected plane.
TL;DR: A modification of the Glees method for silver impregnation of normal and degenerating nervous tissue is described, which since it can be used on batches of slides, is also suitable for histology dass material.
Abstract: A modification of the Glees method for silver impregnation of normal and degenerating nervous tissue is described. It offen geat reliability, evenness of impregnation and since it can be used on batches of slides, is also suitable for histology dass material. The results are illustrated. The procedure for paraffin sections of formalin-Iixed material is as follows: dewaxed and celloidin coated sections are left in 20% silver nitrate solution for 2 hr or until light brown. They are then treated successively in Nauta reducing agent (400 ml distilled water, 45 ml 96% ethanol, 13.5 ml 10% formalin, 13.5 ml 1% acetic acid)—10 min, ammoniacal 5% silver nitrate solution in 80% ethanol (5 g AgNO8 are dissolved in 100 ml 80% ethanol and 25% ammonia solution is added drop by drop until the precipitate clears)—15 min, rinse in absolute ethanol—10 sec. Reduce in two changes of reducing solution (400 ml 10% formalin, 50 ml 96% ethanol, 20 d 1% aretic acid)—10 min, rinse in tap water, fix in 5% sodium thiosulphate—3 min...
TL;DR: A simple technique is described by which retinal whole mounts or slices of brain can be impregnated by the Golgi procedure in a graded manner.
Abstract: A simple technique is described by which retinal whole mounts or slices of brain can be impregnated by the Golgi procedure in a graded manner. The unfixed tissue is laid on a glass slide and covered with a layer of Telfa gauze which is tied in place. Following fixation, the progress of staining, which begins beneath holes in the gauze, is followed by direct observation through the slide. The tissue is impregnated or reimpregnated until the desired results are obtained.