1. |
Nonradioactive in Situ Hybridization with Digoxigenin Labeled DNA Probes |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 59-67
ArnoldN.,
SeiblR.,
KesslerC.,
WienbergJ.,
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摘要:
Nonradioactive in situ hybridization techniques are becoming increasingly important tools for rapid analysis of the topological organization of DNA and RNA sequences within cells. Prerequisite for further advances with these techniques are multiple labeling and detection systems for different probes. Here we summarize our results with a recently developed labeling and detection system. The DNA probe for in situ hybridization is modified with digoxigenin-labeled deoxyuridine-triphosphate. Digoxigenin is linked to dUTP via an 11-atom linear spacer (Dig-[11]-dUTP). Labeled DNA probes were hybridized in situ to chromosome preparations. The hybridization signal was detected using digoxigenin-specific antibodies covalently coupled to enzyme markers (alkaline phosphatase or peroxidase) or to fluorescent dyes. Color reactions catalyzed by the enzymes resulted in precipitates located on the chromosomes at the site of probe hybridization. This was verified by hybridizing DNA probes of known chromosomal origin. The signals were analyzed by bright field, reflection contrast and fluorescence microscopy. The results indicate that the new technique gives strong signals and can also be used in combination with other systems (e.g., biotin) to detect differently labeled DNA probes on the same metaphase plate.
ISSN:1052-0295
DOI:10.3109/10520299209110009
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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2. |
A Two Step Stain for Normal and Leukemic Monocytes Using Two Different Dyes Applied in Sequence |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 68-72
KassLawrence,
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摘要:
A staining procedure for monocytes in specimens of blood and bone marrow was developed. The technique was a two step procedure in which unfixed cells were exposed first to a methanolic solution of C.I. basic blue 54. Next, an aqueous alkaline buffered solution of C.I. basic blue 141 was added to the first staining solution. After staining for 10 min in the solution with two stains, slides or coverslips were washed for 5 sec in pH 5.6 phosphate buffer and drained dry. The cytoplasm of monocytes stained intensely deep purple and frequently nuclei were stained red. Similar staining was not found in other types of normal or abnormal blood and bone marrow cells.
ISSN:1052-0295
DOI:10.3109/10520299209110010
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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3. |
Simultaneous Measurement of DAPI-Sulforhodamine 101 Stained Nuclear DNA and Protein in Higher Plants by Flow Cytometry |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 73-78
UlrichWolfgang,
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摘要:
A 2-step staining procedure is presented for simultaneous measurement of nuclear DNA and protein content in higher plants by flow cytometry. To release nuclei, plant tissues were chopped and stirred in the presence of the DNA specific fluorochrome 4′, 6-diamidino-2-phenylindole (DAPI) and the nonionic detergent Triton X-100. Plant protoplasts were stirred in the DAPI dye solution with the detergent. After a short incubation period a second dye solution containing DAPI and the protein fluorochrome sulforhodamine 101 (SR 101) without detergent was added. Following another incubation, and after filtration through nylon gauze, the highly fluorescent nuclei were analyzed with an impulse cytophotometer. Accurate bivariate DNA-protein histograms were obtained with CV-values of about 2% or less for the 2C-peak of the univariate DNA parameter. The method presented here can be used for basic and applied cytogenetic studies of higher plants, for characterization of subcompartments of the cell cycle phases, or for examination of heterogeneity in plant tissues.
ISSN:1052-0295
DOI:10.3109/10520299209110011
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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4. |
Unknown G0/G1Cell Subpopulation Revealed by Hydrochloric Acid/Acridine Orange Staining |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 79-81
VinogradovAlexander E.,
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摘要:
An unknown cell subpopulation was observed in mouse and rat thymus, spleen and bone marrow cells, as well as in human peripheral blood mononuclear cells (resting and stimulated by PHA) using equilibrium HCl/acridine orange staining. This subpopulation includes cells with decreased green and unchanged red fluorescence. The staining does not affect cells in S- and G2/M-phases. The mechanism and biological meaning of the effect await further investigation.
ISSN:1052-0295
DOI:10.3109/10520299209110012
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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5. |
A Modified Hortega-Globus Stain is Superior to Bielschowsky and Bodian Stains for Demonstrating Neuritic Plaques |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 82-87
BolleL.,
MaurerB.,
JanzerR. C.,
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摘要:
The quantitative assessment of the age-dependent number of neuritic plaques is essential for the diagnosis of Alzheimer type dementia. This study reports the superiority of a modified Hortega-Globus stain compared to Bielschowsky and Bodian stains applied to samples obtained from ten brains of patients with a clinical history of progressive dementia. In two of ten cases only the modified Hortega-Globus stain allowed confirmation of the diagnosis of senile dementia of the Alzheimer type (SDAT). The counts of neuritic plaques in sections stained by other methods were not sufficient to establish the histological diagnosis of SDAT. These results indicate that the choice of the most sensitive staining method is critical for the correct histopathologic diagnosis of the Alzheimer type dementia.
ISSN:1052-0295
DOI:10.3109/10520299209110013
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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6. |
Block-Surface Staining for Differentiation of Starch and Cell Walls in Wheat Endosperm |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 88-97
GlennGregory M.,
PittsMarvin J.,
LiaoKe,
IrvingDelilah W.,
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摘要:
A staining technique for differentiating starch granules and cell walls was developed for computer-assisted studies of starch granule distribution in cells of wheat [Triticum aestivumL.] caryopses. Blocks of embedded caryopses were sectioned, exposing the endosperm tissue, and stained with iodine potassium iodide (IKI) and Calcofluor White. Excessive tissue hydration during staining was avoided by using stains prepared in 80% ethanol and using short staining times. The IKI quenched background fluorescence which facilitated the use of higher concentrations of Calcofluor White. Cell wall definition was improved with the IKI-Calcofluor staining combination compared to Calcofluor alone. The high contrast between darkly stained starch granules and fluorescent cell walls permitted computer assisted analysis of data from selected hard and soft wheat varieties. The ratio of starch granule area to cell area was similar for both wheat classes. The starch granule sizes ranged from 2.1μm3to 22,000μm3with approximately 90% of the granules measuring less than 752μm3(ca. 11μm in diameter). Hard wheat samples had a greater number of small starch granules and a lower mean starch granule area compared to the soft wheat varieties tested. The starch size distribution curve was bimodal for both the hard and soft wheat varieties. Three-dimensional starch size distribution was measured for four cells near the central cheek region of a single caryopsis. The percentage of small granules was higher at the ends than at the mid-section of the cells.
ISSN:1052-0295
DOI:10.3109/10520299209110014
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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7. |
Microwell Cluster for Processing Electron Microscope Sections for Immunocytochemistry |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 98-99
AokiAgustin,
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摘要:
A microwell cluster was manufactured with silicone rubber for incubating thin sections for postembedding electron microscopic immunocytochemistry. The small size of the wells requires only minute amounts (as little as 5μl) of antisera and other valuable immunoreagents. The capillary action of the wells holds the incubation media and grids in place, even if the cluster is turned upside down, thus facilitating safe transport and storage of the sections to be stained. This silicone rubber microwell cluster can also be used as a die to imprint wells in Parafilm sheets to be used for the same purpose.
ISSN:1052-0295
DOI:10.3109/10520299209110015
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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8. |
Epon Resin Infiltration and Immunogold Labelling of Pituitary Secretory Granules after Cryofixation versus Chemical Fixation |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 100-105
EneströmSverker,
KniolaBarbara,
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摘要:
The degree of infiltration of epoxy resin into pituitary secretory granules was evaluated using X-ray microanalysis of the concentrations of chlorine in the epoxy resins. The effectiveness of infiltration was tested after three different tissue preparation techniques: cryofixation + freeze-drying (CF-FD), glutaraldehyde fixation (GF) + chemical dehydration, and no fixation—no dehydration. Signs of marked incomplete infiltration were found in embedded unfixed tissue while the other two techniques showed 80% infiltration. Uneven penetration was seen after CF-FD and GF. The plastic surface demonstrated a mountain-like appearance over the secretory granules after immunocytochemistry of the glutaraldehyde fixed tissue, whereas the CF-FD tissue showed a less furrowed surface. This probably is due to contact with water, which swells those parts of the granules that are unprotected by the plastic embedding medium. Our findings may explain why it is possible to perform immunocytochemistry on Epon embedded tissue.
ISSN:1052-0295
DOI:10.3109/10520299209110016
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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9. |
Sequential and Combined G-Banding for Identifying Breakpoints in Sister Chromatid Exchanges |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 106-109
FusterC.,
MiróR.,
BarriosL.,
EgozcueJ.,
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摘要:
A simple new method is described for obtaining sequential and a combination of differential sister chromatid staining and G-banding in the same metaphase. Using this method the sister chromatid exchanges and chromosome lesion breakpoints can be precisely localized in particular bands of individual chromosomes.
ISSN:1052-0295
DOI:10.3109/10520299209110017
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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10. |
Quality Assurance and Standardization in Immunohistochemistry. A Proposal for the Annual Meeting of the Biological Stain Commission, June, 1991 |
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Biotechnic&Histochemistry,
Volume 67,
Issue 2,
1992,
Page 110-117
TaylorC. R.,
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摘要:
Quality assurance, quality control, proficiency testing, reagent documentation and validation are standard parts of everyday practice in clinical laboratories throughout the United States. Immunohistochemical stains employ reagents and principles in common with immunoenzyme methods utilized in the clinical laboratory. However, immunohistochemistry has not routinely been subjected to similar standardization and quality assurance procedures that manufacturers and pathologists alike have applied to essentially the same techniques in the clinical laboratory environment. The current proposal was invited by the Biological Stain Commission with the charge of incorporating the findings of previous workshops on quality control in immunohistochemistry into a practical design for implementation. The status of quality assurance, quality control and standardization in immunohistochemistry is reviewed and a phased strategy for implementation is proposed.
ISSN:1052-0295
DOI:10.3109/10520299209110018
出版商:Taylor&Francis
年代:1992
数据来源: Taylor
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