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1.
Normand Brière 《Histochemistry and cell biology》1971,28(4):316-323
Summary In the livers of rats fed the azo dye 4-dimethylamino-azobenzene, nucleic acids in connective tissue trabeculae, preneoplastic foci and hepatomas were found to stain intensely with toluidine blue. With the indicator dye bromphenol blue, proteins were observed to stain similarly in these hyperbasophilic tissues but differently from those in surrounding parenchyma.These observations indicate that preneoplastic regions and tumors do not differ from surrounding tissue only by the increased basophilia due to cytoplasmic ribonucleic acid, but also in protein staining. Thus, the change in RNA responsible for hyperbasophila is paralleled by alterations in protein histochemistry. It is suspected that the differential staining of proteins might correspond either to variations in the acidic-basic protein ratios or to the presence of unusual proteins synthesized by such an altered RNA.The tissue similarities in nucleic acid as well as in protein staining observed in the proliferating connective tissue elements and cells undergoing the neoplastic transformation remain an obscure phenomenon.This work was supported by a grant from The Quebec Medical Research Council. 相似文献
2.
Normand Brière 《Histochemistry and cell biology》1971,26(2):113-119
Summary In preneoplastic liver of rats fed the azo dye 4-dimethylaminoazobenzene, certain cellular populations are characterized by cytologic changes typical of tumor cells and appear as the sites of neoplastic transformation. With a basic dye such as toluidine blue, cytoplasmic RNA in preneoplastic foci and hepatomas stains more intensely than in surrounding tissue. In the present study, it was found that when a basic dye (hematoxylin) was combined with an acid dye (tartrazine), these areas stained differentially from the surrounding liver parenchyma. RNAse hydrolysis has shown that such staining properties might be due to the increased proportion of cytoplasmic RNA to components stainable with tartrazine in hyperbasophilic cells, while the surrounding parenchymal cells are probably distinguished by the opposite ratio.It is suggested that the increased basophilia in preneoplastic areas and hepatomas results from the presence of excess RNA and a corresponding decrease in cellular components stained with tartrazine. However, the present method does not permit us to determine whether hyperbasophilia is due to a normal type or an altered form of RNA present in excess.This work was supported by grants from The Quebec Medical Research Council and The René Hébert Foundation. 相似文献
3.
D. W. Menzies 《Biotechnic & histochemistry》1963,38(3):161-163
The red color of nuclei produced in formol-fixed paraffin sections stained with toluidine blue has been investigated by using deoxyribonuclease (DNase), ribonuclease (RNase) and 0.1 M Tris buffer. The action of DNase on formol-fixed material is not fully reliable, but clear-cut when positive. Nuclear basophilia and metachromasia is removed, nucleolar and cytoplasmic RNA is preserved. The picture produced by RNase depends to some extent on the concentration and acidity of the toluidine blue used for subsequent staining. Cytoplasmic RNA is always removed, while the red stain in nuclei usually remains intact. With 0.1% toluidine blue in 1% acetic acid, a nuclear color change from red to pale green is observed. Using this same staining solution, it can be shown that 0.1 M Tris buffer (overnight extraction at 37° C) will remove cytoplasmic RNA but leave intact the nuclear material that stains red. A red to green shift can subsequently be produced by RNase. From this it is deduced that there is a chromatin-associated nuclear RNA fraction which can be removed by the enzyme, but is stable to the buffer solution. 相似文献
4.
Silver staining,immunofluorescence, and immunoelectron microscopic localization of nucleolar phosphoproteins B23 and C23 总被引:21,自引:0,他引:21
Nucleolar organizer region (NOR)-specific silver staining and immunolocalization of nucleolar phosphoproteins B23 and C23 were compared in Novikoff hepatoma ascites cells. Silver staining and protein C23 immunostaining were both localized in the fibrillar shell surrounding the fibrillar center and in the fibrillar center. During mitosis, silver staining and protein C23 were localized at the NORs. Therefore, protein C23 and the silver-staining protein both seem to be associated with rDNA-containing structures (Mirre and Stahl 1981). A comparison of toluidine blue staining specific for RNA and B23 immunostaining demonstrated that protein B23 was associated with RNA-containing regions of the nucleolus and was absent from the fibrillar centers. Localization of these proteins and their functions are discussed in relation to the organization of the nucleolus. 相似文献
5.
A Histochemical Examination of the Staining of Kainate-Induced Neuronal Degeneration by Anionic Dyes
《Biotechnic & histochemistry》2013,88(5):244-254
Anionic dyes, notably acid fuchsine, strongly stain the nuclei and cytoplasm of neurons severely damaged by injury or disease. We provide detailed instructions for staining nervous tissue with toluidine blue and acid fuchsine for optimal demonstration of injured neurons. Degeneration was induced in the hippocampus of the mouse by systemic administration of kainic acid, and the resulting acidophilia was investigated using paraffin sections of the Carnoy-or Bouin-fixed brains. The affected cells were bright red with the toluidine blue-acid fuchsine sequence. Their nuclei were stainable also with alkaline Biebrich scarlet and with the 1,2-naphthoquinone-4-sulfonic acid-Ba(OH)2 method; all staining was blocked by benzil but was relatively refractory to deamination by HNO2. These properties indicated an arginine-rich protein. The nuclei were strongly acidophilic in the presence of a high concentration of DNA (strong Feulgen reaction), and acidophilia could not be induced in normal neuronal nuclei by chemical extraction of nucleic acids. The cytoplasmic acidophilia of degenerating hippocampal neurons was due to a protein rich in lysine (extinguished by alkalinity, easily prevented by deamination, and unaffected by benzil). Stainable RNA was absent from the perikarya of the affected cells, but normal neuronal cytoplasm did not become acidophilic after extraction of nucleic acids. We suggest that kainate-induced cell death is preceded by increased production of basic proteins, which become concentrated in the nucleus and perikaryon. Groups of small, darkly staining neurons were seen in the cerebral cortex in control and kainite-treated mice. These shrunken cells were purple with the toluidine blue-acid fuchsine stain, and were attributed to local injury incurred during removal of the unfixed brain. 相似文献
6.
Summary Using normal rat liver imprints, studies were carried out on the effects of histone extraction and the formation of aldehyde groups from deoxyribose on anisotropic toluidine blue staining of depurinized DNA after sodium bisulfite treatment. The anisotropic effect of bisulfite was found to be determined by binding of bisulfite ions to the aldehyde groups of apurinic acid which, together with free phosphate groups of DNA ensure coparallel attachment of the dye molecules. It was also shown that at pH 5.0 toluidine blue binds with both the phosphate and aldehyde groups of apurinic acid, to give anisotropic staining. 相似文献
7.
J Makovitzky 《Histochemistry》1978,59(1):55-66
In the present study the anisotropic staining of the erythrocyte membrane with 1.9-dimethyl methylene blue and N,N'-diethylpseudoisocyanine chloride was studied and simultaneously compared with the toluidine blue topo-optical staining. The difference between anisotropic toluidine blue and 1.9-dimethyl methylene blue staining, except after KMnO4-oxidation, was only of quantitative nature. On the contrary, striking differences were observed between N,N'-diethylpseudoisocyanine chloride staining, and toluidine blue or 1.9-dimethyl methylene blue staining. Enzymatic and chemical degradation resulted the disappearance of N,N'-diethylpseudoisocyanine chloride staining. Following these treatment membrane birefringence could be restored by aldehyde bisulfate and/or KMnO4-oxidation, while the N,N'-diethylpseudoisocyanine chloride staining was restored only after KMnO4-oxidation. After methylation or acetylation the membrane birefringence disappears, while after KMnO4-oxidation both topo-optical reactions return. The digitonin reaction brought about a rearrangement of the glycocyalyx components. The results draw attention to the spatial orientation of the glycoprotein of the erythrocyte membrane. The role of glycocalyx in the three topo-optical reactions was thus clearly demonstrated. 相似文献
8.
Studies on the molecular arrangement of RNA in tissues with a selective topo-optical reaction of RNA
J. Fischer 《Histochemistry and cell biology》1979,59(4):325-333
Summary Selective, demonstration of RNA in tissues was achieved by treating tissue sections with potassium permanganate followed by bisulfite and toluidine blue at pH. 1.0 (PBT reaction). It is suggested that this reaction is due to aldehyde groups which are formed by the oxidative cleavage of the pyrimidine rings of RNA which can be selectively demonstrated using bisulfite-toluidine blue as the aldehyde reagent.The specificity of the reaction was tested after RNAase treatment, after acid hydrolysis, and on pure RNA droplets. The aldehyde nature of the reacting groups was checked, after permanganate oxidation, by Schiff's leucofuchsin reagent, and by aldehyde blocking reactions.Two types of intracellular molecular arrangement of RNA molecules could be distinguished by polarization optics after application of the PBT reaction: 1) The strong birefringence, dichroism and metachromatic staining of membrane-bound RNA in ergastoplasm of pancreas, liver and plasma cells indicate a linear (planar) molecular order of RNA molecules on the surface of the membranes, and 2) the isotropic, basophilic staining of RNA not organized in membrane structures (Nissl substance, nucleoli) suggest a random distribution of their dye binding sites. 相似文献
9.
Marie Di Berarbino 《Biotechnic & histochemistry》1954,29(5):253-256
Certified toluidine blue (National Aniline Co.). applied to sections of frog blastulae, stained the nuclei light blue and left the yolk platelets either unstained or light blue. Purified toluidine blue (also National Aniline Co.) stained the nuclei a deep blue and the yolk platelets a brilliant pink with deep blue borders. Some of the observations suggest that this difference in staining behavior is due to the presence of an inhibitor in the certified dye, which suppresses the metachromatic staining of the platelets and reduces the intensity of the nuclear staining. Unsuccessful attempts were made to remove the inhibitor by salting out the certified dye and washing it with alcohol or by extracting it with chloroform. Details of these attempts, and of other experiments designed to identify the stainable substrates in the yolk platelets are given in the text. 相似文献
10.
J Fischer 《Histochemistry》1979,59(4):325-333
Selective demonstration of RNA in tissues was achieved by treating tissue sections with potassium permanganate followed by bisulfite and toluidine blue at pH 1.0 (PBT reaction). It is suggested that this reaction is due to aldehyde groups which are formed by the oxidative cleavage of the pyrimidine rings of RNA which can be selectively demonstrated using bisulfite-toluidine blue as the aldehyde reagent. The specificity of the reaction was tested after RNAase treatment, after acid hydrolysis, and on pure RNA droplets. The aldehyde nature of the reacting groups was checked, after permanganate oxidation, by Schiff's leucofuchsin reagent, and by aldehyde blocking reactions. Two types of intracellular molecular arrangement of RNA molecules could be distinguished by polarization optics after application of the PBT reaction: 1) The strong birefringence, dichroism and metachromatic staining of membrane-bound RNA in ergastoplasm of pancreas, liver and plasma cells indicate a linear (planar) molecular order of RNA molecules on the surface of the membranes, and 2) the isotropic, basophilic staining of RNA not organized in membrane structures (Nissl substance, nucleoli) suggest a random distribution of their dye binding sites. 相似文献
11.
Sperm chromatin alteration is an important feature that can affect fertility of the male rabbit. This study compared toluidine blue staining with Feulgen reaction (as methods for evaluating chromatin alteration) and investigated the relationship between sperm morphology and chromatin alteration. Seven hundred rabbit ejaculates of animals with unknown fertility were used. Primary and secondary morphological sperm abnormalities were evaluated in semen smears with phase-contrast microscopy. Chromatin alterations were evaluated in semen smears stained with toluidine blue (pH 4.0 and 5.0) and with the Feulgen reaction. While the three methods were equally efficacious for identification of chromatin alterations, toluidine blue staining was more appropriate to characterize the intensity of chromatin alterations. The correlation between primary sperm defects and chromatin alteration was high and positive, suggesting that sperm chromatin structure affected sperm head morphology. The correlation between secondary sperm defects and chromatin alteration was also positive, but lower. The final chromatin compaction occurs in the epididymus, where secondary sperm defects originate. Therefore, the causes of secondary sperm defects could also intervene with final chromatin compaction. In summary, the toluidine blue stain was an effective means of evaluating the sperm chromatin alteration in rabbit spermatozoa. 相似文献
12.
Using normal rat liver imprints, studies were carried out on the effects of histone extraction and the formation of aldehyde groups from deoxyribose on anisotropic toluidine blue staining of depurinized DNA after sodium bisulfite treatment. The anisotropic effect of bisulfite was found to be determined by binding of bisulfite ions to the aldehyde groups of apurinic acid which, together with free phosphate groups of DNA ensure coparallel attachment of the dye molecules. It was also shown that at pH 5.0 toluidine blue binds with both the phosphate and aldehyde groups of apurinic acid, to give anisotropic staining. 相似文献
13.
不同固定液和染色方法显示山羊子宫内肥大细胞效果的比较 总被引:1,自引:0,他引:1
目的探讨用不同固定液和染色方法对显示处于间情期山羊子宫肥大细胞的影响。方法用四种不同的固定方法,应用改良甲苯胺蓝(MTB)染色法和阿尔辛蓝-番红花红(AB-S)染色法显示处于间情期山羊子宫肥大细胞。结果山羊子宫组织采用Carnoy氏液固定,MTB和AB-S染色对所有的肥大细胞均可获得良好的染色反应,但10%中性福尔马林,4%多聚甲醛,Bouin氏液固定的组织仅有少量肥大细胞着染。结论MTB和AB-S染色法均是山羊子宫肥大细胞良好的染色方法。 相似文献
14.
STUDIES ON NUCLEIC ACID METACHROMASY : II. Metachromatic and Orthochromatic Staining by Toluidine Blue of Nucleic Acids in Tissue Sections 总被引:2,自引:1,他引:1
Acrolein-fixed, polyester wax-embedded tissue sections showed excellent preservation of light microscopic architecture and, when stained with toluidine blue, intense color contrast between DNA, which stained orthochromatically, and RNA, which stained metachromatically. This method has practical value for differentiating DNA from RNA in the same section. The color contrast was impaired by substituting formaldehyde for acrolein or paraffin for polyester wax, and was negligible in tissues fixed in formaldehyde or Carnoy's fluid and embedded in paraffin. Quality of structural preservation paralleled degree of color contrast. Metachromatic staining can be analysed, by the quantitative parameters of Bradley and colleagues, to provide inferences regarding the conformation of biopolymers in tissue sections. Comparison of the nucleic acid color contrasts in toluidine blue-stained sections with titrations of fixative-treated nucleic acids against toluidine blue in solution indicated a greater difference in conformation between DNA- and RNA-protein in acrolein-polyester sections than between acrolein-treated free DNA and RNA in solution. This is supported by recent evidence that the conformation of ribosomal RNA is quite different in whole ribosomes from that assumed by the same RNA free in solution. The acrolein-polyester method may enhance color contrast by providing superior preservation of ordered nucleoprotein conformations. 相似文献
15.
B. de Campos Vidal 《Histochemistry and cell biology》1979,60(3):309-316
Summary When polytene chromosomes are subjected to a clupein treatment, their properties of basophilia and anisotropy are affected. The basophilia is deeply reduced, except in the nucleolar zones, puffs and sites of RNA accumulation. On the other hand, the chromosome birefringence increases. The phenomenon of anomalous dispersion of birefringence usually observed on polytene chromosomes stained with toluidine blue solutions turns into a normal negative dispersion of birefringence, when staining is preceded by clupein treatment. It is concluded that the clupein molecules attach orderly and preferentially to sequential DNA phosphates unbound to chromosome proteins, accentuating DNA anisotropic characteristics. The clupein molecules appear not attaching to RNA phosphates. 相似文献
16.
B de Campos Vidal 《Histochemistry》1979,60(3):309-316
When polytene chromosomes are subjected to a clupein treatment, their properties of basophilia and anisotropy are affected. The basophilia is deeply reduced, except in the nucleolar zones, puffs and sites of RNA accumulation. On the other hand, the chromosome birefringence increases. The phenomenon of anomalous dispersion of birefringence usually observed on polytene chromosomes stained with toluidine blue solutions turns into a normal negative dispersion of birefringence, when staining is preceded by clupein treatment. It is concluded that the clupein molecules attach orderly and preferentially to sequential DNA phosphates unbound to chromosome proteins, accentuating DNA anisotropic characteristics. The clupein molecules appear not attaching to RNA phosphates. 相似文献
17.
Administration of
-galactosamine to rats produces inhibition of liver nuclear RNA synthesis and associated alterations in the structure of the nucleolus. Polyacrylamide gel electrophoretic analysis of liver nuclear RNA from galactosamine-treated rats has shown the virtual complete absence of ribosomal RNA (rRNA) precursor molecules at a time when the nucleolus consists solely of a dense fibrillar core devoid of granules. No evidence for an artefactual, preferential breakdown of nuclear RNA during extraction could be obtained from either 2.7 or 8% acrylamide gels. Furthermore, the almost complete cessation of nuclear RNA synthesis makes the possibility of there being rapid synthesis and degradation of ribosomal precursor molecules in vivo unlikely. With toluidine blue stains for RNA with nuclei isolated from galactosamine-treated animals, the large, brightly staining area associated with the normal nucleolus was not seen. On the basis of these observations, it is concluded that an RNA-depleted nucleolus appears fibrillar. It is suggested that the fibrillar material of a normal nucleolus may not itself be RNA even though this region does contain RNA precursor molecules. 相似文献
18.
鸡中枢淋巴器官肥大细胞的组织化学与形态学 总被引:7,自引:0,他引:7
对哺乳动物的,特别是啮齿动物和人类肥大细胞已有了比较深入的研究,
但关于家禽肥大细胞的研究很少.本研究旨在阐明鸡中枢淋巴器官中肥大细胞的组织化学与形态学特征.本研究证实Carnoy
氏液是鸡肥大细胞的优良的固定液,而中性缓冲福尔马林(NBF)
却阻断了大多数肥大细胞的着染力.甲苯胺蓝和阿尔新蓝是鸡肥大细胞的良好的染料,但阿尔新蓝能使更多的肥大细胞着染,虽然其也可使杯状细胞着染.作者的一种新的染色法,
长时间阿尔新蓝染色(LAB-S)可用于NBF固定的组织中肥大细胞的染色,因为其着染的细胞数与Carnoy
氏液固定甲苯胺蓝染色的细胞数无显著差异(P<0.001).在胸腺髓质中见有大量的肥大细胞,而胸腺皮质仅可见个别肥大细胞位于血管周围及小叶间结缔组织中.腔上囊的皮质与髓质中很少见有肥大细胞.肥大细胞有血管周围分布的倾向,但一个有趣的发现是血管内偶尔也有个别肥大细胞.电镜下可见肥大细胞的胞浆颗粒内充满无定形的颗粒状基质,但其电子密度有的较高,有的较低.少数胞浆颗粒内有旋涡状及网状亚微结构.但未见有人类肥大细胞胞浆颗粒内特征性的晶格状和卷轴状的亚微结构,也未见到在绵羊肥大细胞中描述过的特殊亚微结构. 相似文献
19.
Porphyromonas gingivalis, one of the etiological agents of periodontitis, can be killed by red light in the presence of toluidine blue. The purpose
of this study was to determine whether this light-induced killing was accompanied by changes in the fluidity of the organism's
cytoplasmic membrane. A suspension of the organism was exposed to red light in the presence of toluidine blue, and the membrane
fluidity was monitored spectrofluorimetrically by using the membrane probe trimethylammonium diphenyl hexatriene. The fluidity
of the organism's cytoplasmic membrane was found to decrease significantly during lethal photosensitization, and this was
accompanied by membrane condensation and vacuolation of the cells. Although changes in membrane fluidity are often attributable
to lipid peroxidation, malonaldehyde (a product of lipid peroxidation) was not detectable. The disruption of membrane functions
associated with a decreased membrane fluidity may contribute to the bactericidal effect of light-activated toluidine blue.
Received: 12 October 2001 / Accepted: 7 December 2001 相似文献
20.
胶原蛋白/BMP复合材料的制备和成骨性能研究 总被引:6,自引:0,他引:6
以胶原膜(含87.5 mg I型胶原蛋白)为载体, 复合3.5 mg rhBMP-2(人基因重组骨形成蛋白-2), 制备胶原蛋白/BMP复合材料。复合材料首先在兔背阔肌中埋置, 预构新生骨组织, 并采用ALP染色、Von Kossa染色和HE染色等观察复合材料的成骨过程和组织形态。然后将形成的新骨组织游离移植修复自体下颌骨体部洞穿性缺损; 并设以胶原为载体的rhBMP-2复合骨修复材料直接修复为对照组, 骨缺损不修复组为空白组。采用X线、抗压强度、硬组织切片、四环素荧光染色、骨形态计量检查, 观察复合材料修复骨缺损的质量和效果。结果表明, 胶原蛋白/BMP复合材料在兔背阔肌中4~6周成骨, 胶原材料于3~5周降解; 成骨过程为是以软骨成骨为主的方式, 新骨形态为编织骨, 可见明显的微血管分布; 游离移植修复自体下颌骨缺损, 6周缺损区为骨性愈合, 与对照组在抗压强度(P = 0.041)、新骨量(P = 0.034)均有显著性差异。胶原蛋白/BMP复合材料在骨骼肌中形成的新生骨组织可作为供骨修复一定范围的骨缺损。 相似文献