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1.
For differentiation of cells of the adenohypophysis, the Niagara blue 4B method requires no special preliminary fixative nor very fresh tissue, and requires no more time than routine hematoxylin-eosin (H-E) staining. The method requires fixation in 10% formalin. After processing to paraffin wax, deparaffinise and hydrate the sections and stain in 1% aqueous Niagara blue 4B solution for 2 min. Stain afterwards with hematoxylin for 1 min then differentiate, wash, dehydrate, clear and mount. This method can be used also for staining old HE slides by removing the covers, applying the Niagara blue 4B and restaining with eosin. The Niagara blue 4B combined with H-E gives the best and most colorful result. This method allows special staining of the adenohypophysis from human post-mortem material to become routine. 相似文献
2.
Some staining properties of 10 anionic disazo dyes are clarified by comparison with previous chromatographic analysis. Trypan blue contains both blue and red components and the purified blue fraction displays no color shifts in tissue sections. Evans blue, Niagara blue 2B, Niagara sky blue, Niagara sky blue 4B and Niagara sky blue 6B generally resemble trypan blue. Congo red is a metachromatic dye and the only known example among anionic dyes of established purity whose color shows shifts in tissue sections and also in solutions with certain basic compounds. Other red dyes (Congo corinth, trypan red and vital red) are not metachromatic. The red dye impurity of trypan blue selectively stains nuclei which are pycnotic, degenerating or undergoing no further division. This reaction is apparently related to basic protein content. Other reactions of the red fraction of trypan blue (mammalian erythrocytes, blood plasma) are not fully explained on this basis. 相似文献
3.
Thin-layer chromatography will resolve impurities in commercial dyes, and will do so much faster than paper chromatography. Solvent systems consisting of (a) n-propanol: n-butanol: NH 4OH (conc.): H 2O—4:4:1:1; (b) n-propanol: NH 4OH (conc.): H 2O—8:1:1 on silica gel G plates; and (c) n-propanol: NH 4OH (conc.): H 2O-7:2:1 on Adsorbosil plates were found to be the most effective. Dyes studied were azure A, azure B, azure C, methylene blue, toluidine blue O, thionin, pyronin B, pyronin Y, methyl green, crystal violet amido black 10B and buffalo black (NBR). 相似文献
4.
TO determine the amount of K 2Cr 2O 7 required to produce optimal Giemsa type staining, six 1 g amounts (corrected for dye content) of zinc methylene blue were oxidized with graded quantities of K 2Cr 2O 7 to produce 4, 8, 12, 16, 20 and 24% conversion of methylene blue to azure B. These were heated with a blank control 15 minutes at 100 C in 60-65 ml 0.4 N HCI. cooled, and adjusted to 50 ml to give 20 mg original dye/ml. Aliquots were then diluted to 1% and stains were made by the “Wet Giemsa” technic (Lillie and Donaldson 1979) using 6 ml 1% polychrome methylene blue, 4 ml 1% cosin (corrected for dye content), 2 ml 0.1 M pH 6.3 phosphate buffer, 5 ml acetone, and 23 ml distilled water. The main is added last and methanol fixed blood films are stained immediately for 20-40 min.
For methylene blue supplied by MCB 12-H-29, optimal stains were obtained with preparations containing 20 and 24% conversion of methylene blue to azure B. With methylene blue supplied by Aldrich (080787), 16% conversion of methylene blue to azure B was optimal. Eosinates prepared from a low azure B/methylene blue preparation selected in this way give good stains when used as a Wright stain in 0.3% methanol solution. However, when the 600 mg eosinate solution in glycerol methanol is supplemented with 160 mg of the same azure B/methylene blue chloride the mixture fails to perform well. The HCI precipitation of the chloride apparently produces the zinc methylene blue chloride salt which is poorly soluble in alcohol. It appears necessary to have a zinc-free azure B/methylene blue chloride to supplement the probably zinc-free eosinate used in the Giemsa mixture. 相似文献
5.
Maillet's OsO 4-ZnI 2 fixation staining can be combined with a subsequent counterstaining by Alcian blue or aldehyde fuchsin to demonstrate neurosecretory cells in addition to cytological details of the nerve tissue. This technic has been applied to various annelids: Eisenia foetida (Oligochaeta), Erpobdella octoculata (Achaeta) and Nereis diversicolor (Polychaeta). The material is fixed in a 1:4 mixture of 2% OsO 4 and 3% ZnI 2 for 15 nr, embedded in paraffin, sectioned at 5 μ and the sections alternatively mounted on two glass slides. One of these is oxidized by a solution of 0.3% KMnO 4 acidified by 0.6% H 2SO 4 and counterstained with 1% Alcian blue, pH 0.2, the other one is mounted in balsam. The two preparations may then be compared to locate the neurosecretory cells among the other neurons shown on a slide treated only by the OsO 4-ZnI 2. Secretory cells are not stained by Maillet's reagent; except for their Golgi bodies and their cellular and nuclear membranes. The zone of grains which is generally strongly stained by the Alcian blue takes a yellowish hue from the OsO 4-ZnI 2 fixation. This method could be successfully applied to the histological controls in regeneration experiments. In these last ones, we must simultaneously observe the regeneration of the nervous fibres and the possibility of intervention of neurosecretory elements. 相似文献
6.
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 plychrome methylene blue) using an acetate-formate mixture as eluent. In column B, on an Amberlite polyineric adsorbent (XAD-2) the acetate-formate anions are exchanged for chloride. Regeneration of both columns is possible: KMnO 4, Na 2S 2O 4 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. 相似文献
7.
Subcutaneous injections of 0.25% saline solutions of two dis-azo textile dyes, calcodur pink 2BL, C. I. 353, also known as benzo fast pink 2BL and amidine fast rose 2BL, and a blue dye, dianil blue G, C. I. 508, were made on alternate days on albino rats for one week. The blue dye is closely similar to Niagara blue 4B, C. I. 520, and dianil blue R, C. I. 465. Staining reactions were much like those of other vital blue disazo dyes. Although the pink dye exhibited a similar staining pattern, there were notable differences. The tissues of most glands were stained pink or red. Nuclei of the tubular epithelial cells of the kidney contained red granules as did the cytoplasm of the Kupfer cells. Most unusual was the bright red staining of the elastica interna of medium and large sized arteries. 相似文献
8.
Several samples of J.S.B. stain (Jaswant Singh and Bhattacharjee, 1944) solution 1 (polychrome methylene blue) were prepared with 3-8 hr for dichromate-acid oxidation and addition of varying quantities of Na 2HPO 4 buffer for pH adjustment. Storage under severe tropical conditions and periodical checks by staining Plasmodium cynomolgi smears revealed that staining solutions oxidized 6-7 hr with a final pH of 7.8 gave optimum results. Some precipitation of azures, due to heat after 5 mo, adversely affected the quality of staining solutions, while cooler storage conditions were most favorable. Spectrophotometric and chromatographic studies indicated that the J.S.B. solution 1 was composed of blue and purple components, corresponding to higher methylene azures with methylene blue and thionin with allied products respectively. 相似文献
9.
The described technique, based upon a one-step Mallory-Heidenhain stain, can be applied as a routine stain for glutaraldehyde or OsO 4 fixed, Epon embedded tissues of various organs. The technique consists of a short treatment of the sections with H 2O 2, a nuclear staining with celestine blue B and a final staining in a modified Cason's solution. The different tissue and cell components are displayed as follows: dark brown nuclei, yellow cytoplasm, red collagen fibers and blue elastic' fibers. Intra cytoplasmic components as glycogen and mucus are stained respectively blue and violet, whereas other inclusions such as leucocyte granules are colored orange to red. 相似文献
10.
The granules of islet B cells show an intense β metachromasia when paraffin sections of pancreas fixed in Bouin's fluid or formalin are dipped for 1 min in a 0.1% aqueous solution of toluidine blue O 2 buffered to pH 6.0 with acetate or phosphate. This reaction provides a quick method for surveying the condition of B cells in experimental work. A weak staining is observable at pH 4.5 and becomes distinct at pH 5.5-6.0. Oxidation of sections (0.25% KMnO 4 in 0.5% H 2SO 4, for 1 min, recommended) prior to staining intensifies the metachomatic reaction conspicuously. The metachromatic substance could not be demonstrated after fixation in either ethanol or acetone. It corresponds to the aldehyde fuchsin-positive and pseudoisocyanin-metachromatic substance in its occurrence and distribution in the B cells, as shown by different physiological states of various animals, including fasted and glucose-administered guinea pigs. It is thought to be topographically coincident but not necessarily identical to insulin. 相似文献
11.
Safranin is diazotized by using the customary molar ratio—dye, 1:HCl, 3:NaNO 2l. Partly oxidized NaNO 2 can be used, if necessary, by increasing the concentration of its solution enough to cause the normal color change from red to deep blue to occur within 2 min after adding the NaNO 2. To avoid carrying over excess HNO 2 into the alkaline coupling solution, 1 ml of 3% alcoholic urea solution (30 mg) is added for each milliliter excess of 1 N NaNO 2 used. Any free HNO 2 remaining at the end of the diazotization period produces a deep blue violet on starch-KI paper. Prolonged acid washing may be applied after coupling to decolorize cationic dye staining or triazenes. Na 2S 2O 4, TiCl 3 or SnCl 2 may be used to bleach true azo colorations. This decolorization is not limited to newly formed azo compounds with tissue. 相似文献
12.
Column and paper chromatography of four thiazin dyes revealed both inorganic and organic impurities. In thionin, azure A, azure B and methylene blue, sodium and other metal cations were found as inorganic impurities. The analysis for organic impurities revealed that the dyes were mixtures; specifically each dye contained one or more of the other dyes as impurities. Inorganic impurities were detected by ashing the dyes in the presence of H 2SO 4 and chromatographing the sulfate salts on paper. They were removed by filtration through ion exchange resins. Organic impurities were detected by paper chromatography and removed by column chromatography on Woelm's neutral alumina. 相似文献
13.
After testing various procedures (amidoblack 10B, acid fuchsin-methyl blue, Luxol fast blue MBS-phloxine, toluidine blue O, Jams green B and pinacyanol), three stains can be recommended for staining both types of mitochondria (globose and threadlike) in the cells of Saccharomyces cerevisiae: (1) 0.1% solution of amidoblack 10B in citrate buffer (pH 3.0) for 10 min; (2) 0.01% solution of toluidine blue O in phosphate buffer (pH 6.0) for 30 min; (3) 0.01% solution of Janus green B in distilled water (pH 5.6) for 30 min. The latter stain is most specific because its staining reaction depends upon the action of the mitochondrial enzyme cytochrome c oxidase. Yet, low concentrations and short incubation periods must be applied to avoid poisoning of the cell metabolism. 相似文献
14.
The action of K 2Cr 2O 7, Ag 2O, KMnO 4, HgO and NaIO 3 in polychroming methylene blue is explored. The last two have no action in neutral or acid methylene blue solutions. With the other three reagents the amount of polychroming, as measured by the shift in the absorption spectrum, is roughly proportional to the amount of oxidant used. Various lots of methylene blue produce similar products with similar proportions of K 2Cr 2O 7. With similar quantities of this reagent similar products are produced by polychroming at 100°, 80°, 70° or 60° C. At 100° C. the action of K 2Cr 2O 7 or of Ag 2O appears to be completed in 15 minutes. In K 2Cr 2O 7 polychroming, H 2SO 4 can be substituted for HCl, and subsequent BaCO 3 neutralization removes the salts formed and prevents accidental alkali polychroming. K 2Cr 2O 7 polychroming produces products with narrower absorption bands than alkali polychroming. 相似文献
15.
The ability of 17 inorganic compounds (POCl 3, PSC1 3, PC1 3, P 2O 5, P 2S 5, P 4S 3, P 4S 7, PC1 5, Sb 2O 5, As 2O 5, BiOC1 2, SeOC1 2, SO 2C1 2, Sb 2S 5, VOC1 2, SiC1 4 and CrO 2Cl 2) dissolved in pyridine or 2,2,4-trimethyl pentane, to enhance subsequent staining of tissue components with toluidine blue, phosphotungstic acid-hematoxylin ( PTAH), leukofuchsin, and dihydroxydinaphthyl-disulfide ( DDD) was studied. Eight of these compounds were also tested for ability to enhance staining with Alcian blue 8GN and Luxol fast blue MBS. Nine of the 17 compounds produced increased staining of certain tissue components with leukofuchsin, 13 with toluidine blue, 16 with PTAH, and 16 with DDD. The results suggest additional approaches to identification of tissue entities by induced metachromatic basophilia and leukofuchsin positivity as well as by the other stains studied, and also suggest a number of hitherto unstudied modes of reaction between the dyes used and reactive groups of tissue components. Many reactions of the compounds tested, with reactive groups known to be present in tissue components, are basecatalyzed, so that choice of solvent can influence the results obtained. 相似文献
16.
ABSTRACT. Alcian blue acts as a secretagogue and chemorepellent in a variety of unicellular eukaryotes. We report that alcian blue stimulates flagellar excision and induction of RNA encoding flagellar proteins in Chlamydomonas reinhardtii . Flagellar excision by alcian blue is dependent on extracellular Ca 2+ and is blocked by La 3+, ruthenium red, and neomycin, and so is similar to flagellar excision by acid shock. However, the adf-l mutant excises its flagella following alcian blue treatment, but not following acid shock, thus genetically distinguishing alcian-blue-induced excision from acid-shock-induced excision. Wild-type, but not adf-1, cells regrow their flagella in the continued presence of alcian blue. Wild-type cells that regrow flagella in the presence of alcian blue fail to excise their flagella in response to either increased concentrations of alcian blue or to acid shock. Alcian blue treatment of cells also induces RNA encoding flagellar components, but in a manner distinct from other means of stimulation. These results suggest that treating Chlamydomonas with the secretagogue alcian blue initiates a Ca 2+ influx pathway and that prolonged treatment with alcian blue desensitizes the acid-shock-activated Ca 2+ influx pathway to acid treatment. Alcian blue will thus be a useful excitatory ligand in future studies of receptor-mediated Ca 2+ signaling in the Chlamydomonas flagellar regeneration system. 相似文献
17.
The addition of buffer mixtures to toluidine blue O, thionin and new methylene blue N improves their use in the staining of mucus after formaldehyde fixation. Overstaining is minimi The buffer mixtures used consisted of variable proportions of M/10 citric acid and M/5 anhydrous Na 2HPO 4 in 25% methanol. Connective tissue mucus stained satisfactorily with these dyes at a buffer pH range of 3.4 and 3.95 and epithelial mucus at a range of 2.2 to 3.95. The corresponding ratios of M/10 citric acid: M/5 Na 2HPO 4 are 16:4 to 14:6 and 20 A to 14:6 respectively. 相似文献
18.
本试验采用发光二极管(LED)精准调控光源环境,以‘改良大龙茄’为试材,在前期探明红/蓝组合光(R/B)=3/1有利于培育茄子壮苗基础上,以R/B=3/1(9/3,CK 1)和LED白光(W,CK 2)为对照,分别设定LED红蓝白组合光为: R/B/W=3/1/1(9/3/3,T 1)、R/B/W=9/3/8(T 2)、R/B/W=3/1/6(9/3/18,T 3)和R/B/W=3/1/16(9/3/48,T 4),研究其对茄子幼苗生长、根系发育及定植后茄子产量和品质的影响,以期为茄子集约化高效育苗和设施生产补光提供理论依据和技术指导.结果表明: R/B/W=9/3/8组合光处理下,茄子幼苗株高、茎粗、壮苗指数、地上部干重、根干重、根系发育,以及定植后茄皮花青素、类黄酮含量和前期产量均较高;R/B/W=3/1/1组合光处理下,幼苗根冠比、定植后茄皮总酚和果肉中可溶性糖含量较高;而R/B/W=3/1/6组合光处理下,全株叶面积及定植后茄子果肉中游离氨基酸含量较高;R/B/W=3/1/16组合光处理下,定植后茄子果肉中可溶性蛋白含量较高.不同红蓝白组合光可不同程度提高茄子幼苗质量及定植后茄子前期产量和品质,以R/B/W=9/3/8组合光效果最显著. 相似文献
19.
It is suggested that celestine blue B can stain as a colloidal dispersion, the nuclear specificity of which is controlled by the pH. The staining solution is prepared by adding 0.5 ml of concentrated H 2SO 4 to 1 gm of celestine blue B and dissolving the resultant granular mass in 100 ml of 2.5% ferric alum containing 14 ml of glycerol. Sections of amphibian, avian, and mammalian tissue placed for 1 min in this solution and then rinsed in water show as sharp nuclear staining as that usually produced by hematoxylin. A wide variety of fixatives is permissible. Overstaining is not possible within reasonable limits of exposure and no differentiation nor bluing is required. Both the staining solution and stained slides are stable. 相似文献
20.
1. 1.|The raccoon dogs frequently basked in spring while keeping their dark chest area towards the sun. The importance of this behaviour for the thermal balance was examined by using a cylinder model, and the results were compared with that of the blue fox which has pale chest and no basking behaviour. 2. 2.|With no external radiation source, cooling rates of blue fox and racoon dog models were almost equal, while in the sunshine with the chest area towards the sun, raccoon dog gained and blue fox lost heat. 3. 3.|In the same sunshine, the raccoon dog lost heat if its back area was towards the sun in comparison with the situation when the chest area was towards the sun. 4. 4.|Temperatures at the skin level were much higher for sun-exposed raccoon dogs than blue foxes especially on the chest area. 5. 5.|It is concluded that the hair coat structure of the raccoon dog is especially favourable for trapping heat from the sun, and with postural adjustments the animal takes maximal advantage of this free heat.
Author Keywords: Raccoon dog; coat colour; basking behaviour; blue fox; thermal radiation 相似文献
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