摘要:

Staining of etched sections for light microscopy is described. Azan staining was successful after treatment with potassium dichromate and the use of concentrated dye solutions. To remove osmium for hematoxylin-eosin staining, removal by reduction with ferrocene was used instead of oxidation. Highly selective differentiation after hematoxylin staining was achieved usingp-toluenesulfonic acid-DMSO. To enhance eosin staining, a 2-bromoethylamine link between eosin and the tissue was used. Ferrocene also facilitated counterstaining of nuclei with hematoxylin after the PAS reaction. Periodic acid-methenamine silver staining was carried out without modification.

ISSN:1052-0295    

DOI:10.3109/10520299509108317

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Eight fluorescent dyes were tested for staining the spores or mycelia of six fungi and for their translocation into new growth when the preloaded spores or mycelia were incubated on agar coated coverslips. The dyes studied were Cellufluor, Nile red, fluorescein diacetate (FDA), carboxyfluorescein diacetate (CFDA), chloromethylfluorescein diacetate (CMFDA), aminochloromethyl coumarin (CMAC), and the carbocyanines DiIC18(3) and DiOC18(3). The fungi on which the dyes were tested includedBotrytis cinerea, Fusarium oxysporumf.sp.lycopersici, Idriella bolleyi, Pythium oligandrum, Sclerotium cepivorumandTrichoderma. harzianumMost of the fluorochromes gave good initial staining of mycelia or spores; however, FDA fluorescence faded rapidly during excitation, making it impractical for use. Also, the spores and mycelia ofB. cinereaandT. harzianumsometimes gave weak fluorescence with Nile red, and the spores and mycelia ofI. bolleyigave unusually weak fluorescence with Cellufluor. There were other variations of staining among the different dye/fungus combinations, but each fungus showed strong fluorescence with at least one dye. Cellufluor, CMFDA, CMAC and, to a lesser extent, CFDA and Nile red, were efficiently translocated into new growth from preloaded spores or mycelia, whereas FDA, DiIC18(3) and DiOC18(3) were not. The extent of translocation ranged from 0.1 to 1.2 mm in germ tubes arising from spores, and from 0.9 to 9.2 mm in mycelia extending from dye-loaded agar blocks. The findings suggest that fluorescent dyes could be used as markers or tracers in studies of fungal growth and differentiation.

ISSN:1052-0295    

DOI:10.3109/10520299509108318

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

To investigate the time course of mineralization in undecalcified dental tissues, calcein-and tetracycline-labeled rat maxillary molar sections were stained with Villanueva bone stain en bloc, embedded in methyl-methacrylate (MMA), ground to 50μm thickness, and observed by confocal laser scanning microscopy (CLSM). This method allowed observation of dental structures including odontoblasts, pulp cells and periodontal ligament, and dentinal tubules and enamel rods at high resolution; labeled enamel, dentine, and cementum could be observed simultaneously regardless of section thickness. CLSM permitted simultaneous observation of both the components of calcified tissue and the cellular components of dental tissues, and assessment of the mineralization time course of hard tissues labeled by tetracycline or calcein. The technique is useful for both assessing the elements composing dental structure and observing the histological dynamics by which dental structure develops.

ISSN:1052-0295    

DOI:10.3109/10520299509108319

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

A staining method is described using thionin, for undecalcified deacrylated bone sections. RNA is stained purplish violet, allowing still active osteoblasts to be distinguished from lining cells. Staining intensity of mineralized bone is related to the degree of mineralization. Mineralizing fronts and cement lines are visualized clearly. Lamellae show an alternate pattern. Histomorphometric parameters such as osteon thickness and interstitial bone thickness can be measured without using polarized light. The mineralizing front can be assessed and expressed as a percentage of the osteoblast-covered interface between osteoid and mineralized bone. The stain is also useful for qualitative assessment of metabolic bone disease. Thionin stained sections can be kept for at least one year when stored hi the dark at 7 C.

ISSN:1052-0295    

DOI:10.3109/10520299509108320

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

In this report we describe a simple and rapid staining technique for cartilage and bone embedded in Araldite. Semitbin sections of embryonic vertebrae obtained from 15 to 17 day mouse fetuses were stained using an aqueous solution 0.25% with respect to methylene blue, 0.25% with respect to azure A, and 0.5% with respect to Na2CO3, then counterstained with 1% aqueous pararosaniline chloride (MAP). Results were compared with toluidine blue stained sections. MAP permitted good discrimination of developmental stages of both cells and extracellular matrix within vertebral ossification centers during endochondral ossification. The technique is simple, rapid and applicable to plastic embedded sections, and can be used prior to ultrastructural examination.

ISSN:1052-0295    

DOI:10.3109/10520299509108321

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

The calcium distribution in the ampullary electroreceptor and the type B electrore-ceptor organ (gymnarchomast) ofGymnarchus niloticus(Gymnarchidae) and in the tuberous organ ofApteronotus leptorhynchus(gymnotidae) was studied. Endogenous calcium appeared as electron-dense precipitates when the cutaneous organs were pre-fixed with phosphate-buffered glutaraldehyde and postfixed with osmium tetroxide plus potassium bichromate. Calcium precipitates were localized in both intracellular compartments of sensory cells and afferent nerve fibers. In contrast to sensory cells, small amounts of calcium precipitates were found in the cytoplasm of accessory cells. In sensory cells, electron-dense deposits were apparent mainly in synaptic vesicles near synaptic ribbons, inside vacuoles of the endoplasmic reticulum, and between the layers of the nuclear membrane. Very few deposits were found in mitochondria. Precipitates were also observed within the axons of afferent nerves and between the layers of the myelin sheath. The synaptic cleft was devoid of calcium. Calcium deposits have a specific cellular distribution in electro-receptor organs of teleost fish.

ISSN:1052-0295    

DOI:10.3109/10520299509108322

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

To investigate the relationship of oncogene analysis to morphology, we analyzed K-rasgene mutations by dot-blot hybridization with and without consideration of histological atypias in individual colorectal adenomas. Each of 54 colon polyps were divided into two parts after fixation. One part was used as a mass to assess point mutations; the remaining portion of each polyp was paraffin-embedded, stained with hematoxylin and eosin, and examined for point mutations related to histological atypias. In the first part of our study, K-rasgene mutations at codon 12 were detected in 13 cases (24%). In the second part of our study, 12 cases had distinctly different histological atypias. From each of these 12 cases, two areas, one with higher or one with lower grade atypia in the same polyp were excised to analyze forK-rasgene mutation. Two of these 12 cases (17%) had the mutation in different areas of the same tumor. These two cases contained the mutation only in the areas with higher grade atypia, and only one case added information regarding ras mutation upon microdissection when compared to the entire biopsy. These results suggest that oligonucleotide hybridization can identify the majority of cases containingrasmutations despite regional morphologic variation. Individual cases, however, may contain clonal subpopulations within adenomas with differentrassequences from other regions within the same adenoma.

ISSN:1052-0295    

DOI:10.3109/10520299509108323

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Strains ofRhizoctonia solani, a common soil-borne, pathogenic fungus of plants, are assigned to one of 11 anastomosis groups (AGs) based on the occurrence of imperfect fusions (anastomoses) between hyphae of a non-typed strain and a tester strain of one of the 11 AG's. Imperfect fusion is characterized by the death of one or more cells in each of the hyphae involved in the fusion. Although hyphae from branches of the same strain of JR. solani may fuse with each other (self-fusion), cell death does not occur. Cell death is accompanied by nuclear degradation and granulation, or plasmolysis of the cytoplasm, which often is not visible using bright-field microscopy. When the DNA-binding fluorochrome DAPI (4′, 6-diamidino-2-phenylindole) is used and the hyphal fusions viewed under fluorescence microscopy, no nuclei are observed in fused hyphal cells from two strains of the same AG ofR. solaniBecause DAPI reacts only with living nuclei, lack of staining is presumptive evidence that the fused cells are dead as a result of imperfect fusion. The use of DAPI reduces the time required for making AG determinations compared to standard methods because it eliminates the need to assess cell wall dissolution and cytoplasmic fusion. Also, it is not necessary to trace the hyphae involved in the fusion to their respective origins to ensure that self-fusion has not occurred.

ISSN:1052-0295    

DOI:10.3109/10520299509108324

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

After brief exposure to an aqueous solution of the oxazine textile dye C. I. basic blue 148 following fixation in 37% formalin, 95% ethanol and glacial acetic acid, T helper cell nuclei and cytoplasm in specimens of peripheral blood displayed a deep red-violet color. No other cell in normal blood or bone marrow specimens showed intense staining of this type. The total staining time is 1 min. Basic blue 148 stain is a promising technique for hematology and immunology laboratories as a rapid screening test for T helper cells in blood specimens using a microscopic slide and ordinary incandescent illumination.

ISSN:1052-0295    

DOI:10.3109/10520299509108325

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor