摘要:

A bromodeoxyuridine (BrdU) labeled DNA probe was used for in situ hybridization at the electron microscopic (EM) level. A BrdU labeled DNA probe was hybridized in situ to cryostat sections of paraformaldehyde fixed OCT compound embedded cultured HL-60 cells. After hybridization, some sections were incubated with FITC-conjugated anti-BrdU monoclonal antibody for fluorescence microscopy (FM). and others were embedded in Quetol for electron microscopy (EM). The ultrathin sections of Quetol-embedded specimens were incubated with the anti-BrdU monoclonal antibody and the immunoglobulin: gold colloid. In both FM and EM studies, the signals were concentrated in the rough endoplasmic reticulum. Moreover, some label was arranged from the nucleus to the cytoplasm at the EM level. Relatively simple methods using the BrdU labeled DNA probe for the detection of the defined nucleic acid sequence with reasonable tissue preservation and high resolution are described here. This method may be useful for developmental and disease related studies of specific mRNA in cells and tissues.

ISSN:1052-0295    

DOI:10.3109/10520299309104691

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Formation of crystals in Schiff reagents prepared from SO2gas previously has been reported either soon after preparation, using high dye concentrations and heating, or after long periods of storage at room temperature. With the first type of procedure only a low yield of crystals accompanied by dye precipitation was obtained. Crystallization without dye precipitation took place if the reagent, prepared with pararosaniline base or chloride in a saturated SO2solution, was stored for a sufficient time at room temperature in partly filled flasks. These crystals remained colorless if washed with acid alcohol after being separated by filtration. Schiff reagents layered with paraffin oil or supplemented with 0.1 M hydroquinone took much longer to crystallize, suggesting that crystallizaticn is promoted by the partial oxidation of sulfurous acid to sulfuric acid. A high yield of crystals can be obtained at room temperature after as little as 24 hr by adding 0.04 M of H2SO4to a Schiff reagent prepared with 2% pararosaniline chloride in a saturated SO2solution. A Schiff reagent prepared with only 0.2% of these crystals gives an intense staining in the Feulgen and in the Periodic acid-Schiff reactions.

ISSN:1052-0295    

DOI:10.3109/10520299309104692

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

The effect of various treatments on the Sudanophilia of the mineralizing fronts of hard tissues has been examined. We have shown that a variety of organic solvents. but not all lipid unmasking protocols. expose Sudanophilic lipids at the mineralizing fronts of dentine, enamel matrix, bone and cartilage by the extraction of a substance which is not Sudanophilic. A variety of organic solvents, but not all extraction protocols. abolish Sudanophilia at the mineralizing fronts of bone and cartilage. The present study indicates that only solvent mixtures containing methanol abolished Sudanophilia at the mineralizing fronts of dentine and enamel matrix.

ISSN:1052-0295    

DOI:10.3109/10520299309104693

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

ISSN:1052-0295    

DOI:10.3109/10520299309104694

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor