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《Biotechnic & histochemistry》2013,88(5):259-262
Histochemical 1,2-glycoI cleavage, similar to that obtained with periodic acid and lead tetraacetate, may be obtained with sodium bismuthate. Routinely prepared slide sections, from tissues fixed in 10% formalin, are run down through xylene and graded alcohols to water and then oxidized for three minutes in a 1% sodium bismuthate 20% aqueous phosphoric acid solution. The oxidizing solution must be freshly prepared and used immediately. Following oxidation, sections are rinsed 15 sec. in IN HC1 to remove bismuth pentoxide precipitate, a by-product of the reaction. The sections are then washed in distilled water and placed in leuco-fushsin for 10 min., or in a saturated 30%) alcoholic solution of p-nitrophenylhydrazine for 5 min. or 2,4-dinitrophenylhydrazine for 30 minutes. After staining, the sections are rinsed in 30% alcohol if the nitrophenylhydrazines were used, or in the standard dilute sulfite bath followed by running tap water for 5 min. if leucofuchsin were used. Sections are routinely dehydrated, cleared, and covered. On examination, the sites of 1,2-glycol linkages will be stained violet by leucofushsin or yellow by the nitrophenylhydrazines. 相似文献
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Complete blocking of the Schiff reaction applied after HIO4 oxidation is attained by 1%, 0.5% or 0.2% NaBH4 in 1% Na2HPO4 with a 1 min exposure, 0.1% NaBH4 requires 2 min. KBH4 was also completely effective in the same solvent at 1, 0.5 and 0.25% in 1 min. The solutions deteriorate on standing, so that 0.5% NaBH4 is effective in 1 min at 7 hr but 5 min is needed at 24 hr and is ineffective at 36 hr; 1% KBH4 requires 4 min at 24 hr when fresh, and showed deterioration in 2-4 days. Saturated (0.4%) isopropanol and 1% pyridine solutions required 10-15 min when fresh and showed deterioration in 2-4 days. No satisfactory results were obtained with 80% or absolute dioxan or with methyl cellosolve in times of an hour or less; even a 24 hr exposure was ineffective with 0.2% in 80% dioxan. 相似文献
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Schiff reagents were made by two methods. The first procedure gave a Schiff reagent of pH 1.8-2.4. It was accomplished by passing sulfur dioxide into 0.5% aqueous fuchsin solution at room temperature, stopping at reddish violet, and decolorizing allowed to occur on standing. In another method, 1.5 ml. of 5.6% sulfurous acid was added to 100 ml. 0.5% fuchsin solution and the mixture produced in several hours a colorless Schiff reagent of pH 3. The solution remained unchanged for some weeks when kept stoppered in a refrigerator.
To test these Schiff reagents, histochemical examinations were carried out with Feulgen and McManus reaction in various pH ranges. These experiments showed that the Feulgen reaction was optimum at pH 3, the McManus reaction at pH 2.4. 相似文献
To test these Schiff reagents, histochemical examinations were carried out with Feulgen and McManus reaction in various pH ranges. These experiments showed that the Feulgen reaction was optimum at pH 3, the McManus reaction at pH 2.4. 相似文献
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J. F. Lhotka 《Biotechnic & histochemistry》1955,30(5):235-238
If the standard Soxhlet continuous-extraction apparatus is modified by the addition of a water jacket around thextraction chamber, the temperature of tissue undergoing extraction can be kept safely below damaging limits. Subsequent routine processing and examination of tissues after the cool extraction showed that they were less friable and had fewer structural distortions than those extracted with the uncooled, standard type c apparatus. 相似文献
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A technique for the application of dieca (diethyldithiocarbamate)to plant tissues is described. It is shown to differentiatealmost quantitatively between extracted ascorbic oxidase andpolyphenolase on the one hand and cytochrome oxidase on theother at a concentration of M/5,000. It produces strong, butnot total, inhibitions when applied to barley root-tips. 相似文献
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Thinnest practical slices of bones or teeth are suspended at 4 C in a 10% solution of EDTA in 0.1 M tris buffer brought to pH 6.95 with KOH pellets. The solution is stirred moderately and continuously with a magnetic stirrer until specimens are demineralized. Unwashed specimens, taken directly from the demineralizing fluid, are frozen on a block of CO2 ice, mounted on a tissue carrier and sectioned at 6 μ in a cryostat. Histochemical stains conducted on sections stored in a slide holder enclosed in a plastic bag in a refrigerator for as long as 2 wk were successful. Quantitative studies on the preservation of succinic dehydrogenase showed nearly all of it present in specimens demineralized for 2 days, and approximately 50% remaining in specimens demineralized for 7 days. Qualitative studies with other dehydrogenases indicate that several may be similarly affected. 相似文献
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David Morton 《Biotechnic & histochemistry》1978,53(4):217-223
Four histochemical tests for iron and four procedures for its removal were investigated in regard to their suitability for glycol methacrylate embedded tissues. The HCl-ferrocyanide and chlorate hematoxylin methods were easily modified for plastic sections. The latter does not use iron-containing reagents. Desiderization was complete both after a fifteen minute exposure in 1% Na2S2O4 in 0.1 M acetate-HCl buffer (pH 4.5) and, if an acid method is preferred, after twelve hours in 5% oxalic acid. A six hour treatment in 3.7 N H2SO4 also removed all histochemical iron but was accompanied by a relatively greater loss of tissue basophilia. 相似文献
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John F. Lhotka 《Biotechnic & histochemistry》1952,27(4):213-216
Lead tetra-acetate acts specifically to split the carbon-carbon single bond of the 1,2-glycol linkage to produce aldehyde radicals which may then be demonstrated by means of leucofuchsin, 2,4-dinitrophenlyhydrazine, or p-nitrophenylhydrazine. Routinely prepared slide sections from tissues fixed in 10% formalin are run down to 95% alcohol, rinsed in glacial acetic acid and then treated for 2 minutes in a saturated solution of lead tetra-acetate in glacial acetic acid with 5 g. of potassium acetate added for each 100 ml. of reagent. The sections are then washed in distilled water and placed in leucofuchsin for 10 minutes, or in a saturated 30% alcoholic solution of p-nitrophenylhydrazine for 5 minutes or 2,4-dini-trophenylhydrazine for 30 minutes. After staining, the sections are rinsed in 30% alcohol if the nitrophenylhydrazines were used, or in the standard dilute sulfite bath followed by running tap water for 5 minutes if leucofuchsin were used. Sections are routinely dehydrated, cleared, and covered. On examination, the sites of 1,2-glycol linkages will be stained violet by leucofuchsin or yellow by the nitrophenylhydrazines. 相似文献
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Fresh tissue slices were fixed in 5% formalin containing 0.9% NaCl for 10-20 min and frozen sections therefrom floated for 3 hr at 37°C on an incubating mixture made as follows. Sodium pyrophosphate (Na4P2O7-12H2O), 1.088 gm was dissolved in 20-30 ml of distilled water and to this was added ferric chloride (FeCl3-6H2O), 0.61 gm dissolved in 10-15 ml of water. The precipitate was just dissolved by cautiously adding 5-10% aqueous Na2CO3 solution and the pH adjusted to 7.2 with 1N HCl. The volume was made up to 100 ml and 0.9 gm of NaCl added. Before use, 1 ml of 10% Mg(NO3) was added. After incubation, sections were washed 10-15 min in 0.9% NaCl, then mounted on glass slides and air-dried. When dry, the slides were immersed in 0.9% NaCl containing 0.2-0.5% ammonium sulfide for 2-3 min, then dehydrated rapidly through graded alcohols, cleared, and covered in balsam. Sites of pyrophosphatase activity stained in various shades of green. Acid pyrophosphatase also was histochemically demonstrated by the same principle, excepting that the substrate solution was adjusted to pH 3.7-4.0 with acetate buffer. The pattern of distribution of pyrophosphatase and glycerophosphatase was almost identical. 相似文献
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Summary A new simple method for the histochemical demonstration of enterokinase (enteropeptidase, EC 3.4.4.8) was elaborated. Unfixed cold microtome sections adherent to the slides are covered with agar-gel medium containing trypsinogen (0.5–1 mg per 1 ml), N-benzoyl-DL-arginine--naphthylamide hydrochloride (2–4 mM), and Fast Blue B Salt (0.5 mg per 1 ml) in 0.05 M Tris maleate buffer pH 6.5 with 0.05% CaCl2. After the incubation in a wet chamber at 37° C the slides are chelated in 2% copper sulphate. The method enables a localization on the histological level. The evaluation of the overall staining obtained with this method in sections of the duodenum, jejunum and ileum of guinea pigs, rats, monkeys, dogs and humans reflects well the biochemical quantitative data on enterokinase activity obtained in homogenates of mucosal scrapings of the corresponding gut segments of the same animals.In all animals a proximodistal gradient was found. The highest activities were found in enterocytes covering apical parts of villi in the duodenum. According to the activities in the duodenum the investigated animals can be arranged in the following series: guinea pig (highest activity), monkey, man, rat and dog. 相似文献
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Histochemical demonstration of aminopeptidase 总被引:8,自引:0,他引:8
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This study describes the distribution, proteoglycan properties and protease activity of mast cells from 15 different dog organs. In beagles and mixed breed dogs, staining with Alcian Blue-Safranin O revealed mast cells in all the organs examined. However, their numbers varied and they demonstrated unique localization patterns within some of these organs. Berberine sulphate fluorescence-positive mast cells were observed in the submucosa, muscularis and serosa of the intestines, as well as the tongue and liver (within the connective tissue). Mast cells within the intestinal mucosa were negative for, or demonstrated weak, berberine sulphate staining. Heterogeneity of mast cells in terms of the proteoglycans contained within their granules was further confirmed by determination of critical electrolyte concentrations (CECs). The CECs of mast cells within the connective tissue of several organs, including the intestines (submucosal and muscularis-serosal layers) were all greater than 1.0 M. The results from CEC experiments together with berberine staining indicate that heparin was contained within their granules. Relative to the CECs of mast cells in other organs, mast cells in the intestinal mucosa exhibited lower CECs, suggesting that the proteoglycans within their granules were of lower charge density and/or molecular weight. Although mast cells were classified into two groups by proteoglycans within the granules, enzyme histochemical analysis in beagles revealed three subtypes of mast cells: chymase (MC(C)), tryptase (MC(T)) and dual positive (MC(TC)) cells. There was no correlation between the proteoglycan content and enzyme properties of the mast cell granules. 相似文献