Reference - Detail
| RRC ID | 33960 |
|---|---|
| Author | Davis WP, Janssen YM, Mossman BT, Taatjes DJ. |
| Title | Simultaneous triple fluorescence detection of mRNA localization, nuclear DNA, and apoptosis in cultured cells using confocal scanning laser microscopy. |
| Journal | Histochem Cell Biol |
| Abstract |
We describe a multifluorescence labeling technique for simultaneous detection of mRNA, nuclear DNA, and apoptosis in cultured cells. Digoxigenin-labeled cRNA probes were used to study proto-oncogene expression in rat pleural mesothelial cells undergoing apoptosis following exposure to crocidolite asbestos or hydrogen peroxide (H2O2). Hybridized cRNA probe was detected by immunolocalization with an anti-digoxigenin monoclonal primary and fluorophore-conjugated anti-mouse secondary antibody. Cells undergoing apoptosis were simultaneously identified by the TdT-mediated biotin-dUTP nick-end labeling (TUNEL) method and a streptavidin-conjugated far-red fluorophore, and nuclear DNA was stained with oxazole yellow dimer (YOYO-1). With confocal scanning laser microscopy, we demonstrated increased c-jun mRNA expression within the cytoplasm of both TUNEL-positive and non-apoptotic cells following exposure to either crocidolite asbestos or H2O2. Thus, this technique represents a useful in vivo approach for evaluating apoptosis-associated gene expression with confocal scanning laser microscopy. |
| Volume | 108(4-5) |
| Pages | 307-11 |
| Published | 1997-1-1 |
| DOI | 10.1007/s004180050170 |
| PMID | 9387922 |
| MeSH | Animals Apoptosis* Asbestos, Crocidolite Benzoxazoles / chemistry Carbocyanines / chemistry Cell Nucleus / chemistry Cells, Cultured DNA / analysis* Fluorescence Fluorescent Dyes / chemistry In Situ Hybridization, Fluorescence / methods* Microscopy, Confocal / methods* Proto-Oncogene Proteins c-fos / genetics Proto-Oncogene Proteins c-jun / genetics Quinolinium Compounds / chemistry RNA, Messenger / analysis* Rats Ribonucleases / metabolism Water |
| IF | 3.418 |
| Times Cited | 15 |
| WOS Category | MICROSCOPY CELL BIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| Total number of mentions | 0 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | pBS rcjun-1 (RDB01130) |