NONRADIOACTIVE IN-SITU HYBRIDIZATION TECHNIQUES FOR ROUTINELY PREPARED PATHOLOGY SPECIMENS AND CULTURED-CELLS

被引：14

作者：

GAN, FY ^{[1
]}

LUK, GD ^{[1
]}

GESELL, MS ^{[1
]}

机构：

[1] DEPT VET AFFAIRS MED CTR,DALLAS,TX 75216

来源：

JOURNAL OF HISTOTECHNOLOGY | 1994年 / 17卷 / 04期

基金：

美国国家卫生研究院;

关键词：

BETA-ACTIN; C-MYC; CELL CULTURES; DIGOXIGENIN-LABELING; IN SITU HYBRIDIZATION; ORNITHINE DECARBOXYLASE; PATHOLOGY ARCHIVES; RANDOM PRIMER EXTENSION;

D O I：

10.1179/his.1994.17.4.313

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

We report a modified method for non-radioactive in situ hybridization suitable for semiquantitative analysis of specific mRNA sequences in fixed cultured cells or formalin fixed, paraffin embedded tissue sections. Our modifications include random primer incorporation of digoxigenin-label into DNA probe; specimen treatment with proteinase K; heating of specimen and hybridization solution to 100 degrees C for 10 min followed by hybridization at 42 degrees C for 18 hr; and improved immunologic detection with an antibody against digoxigenin conjugate. Visual results were obtained within 3 days. Probe sizes as large as 1.8 kb resulted in good specific signals with minimal background. Specific histologic localization can be semiquantitated under a brightfield microscope linked to a computer aided densitometry system.

引用

页码：313 / 319

页数：7

共 12 条

[1] INSITU HYBRIDIZATION AT THE ELECTRON-MICROSCOPE LEVEL - LOCALIZATION OF TRANSCRIPTS ON ULTRATHIN SECTIONS OF LOWICRYL K4M-EMBEDDED TISSUE USING BIOTINYLATED PROBES AND PROTEIN-A GOLD COMPLEXES [J].