Optimal prefixation of cells to demonstrate apoptosis by the TUNEL method
| Autor: | Yasuhiko Kiyozuka, Airo Tsubura, Takako Akamatsu, Yogendra Prasad Singh, Hideto Senzaki, Hiroshi Ichiyoshi |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Pathology
medicine.medical_specialty Histology Tissue Fixation Cell Ovary Antineoplastic Agents Apoptosis DNA Fragmentation Cell morphology Pathology and Forensic Medicine Polyethylene Glycols DNA Nucleotidylexotransferase Cytology medicine Tumor Cells Cultured Humans Biotinylation Cisplatin Ovarian Neoplasms TUNEL assay Ethanol business.industry General Medicine DNA Neoplasm Molecular biology Cystadenocarcinoma Serous medicine.anatomical_structure Cancer cell Female business medicine.drug |
| Zdroj: | Acta cytologica. 43(3) |
| ISSN: | 0001-5547 |
| Popis: | OBJECTIVE To determine the optimal fixation method for cultured human ovarian cancer cell line SHIN-3 to document cisplatin-induced apoptosis by the terminal deoxynucleotidyl transferase-mediated biotinylated dUTP nick end-labeling (TUNEL) assay. STUDY DESIGN Cisplatin-treated cancer cell suspensions were (1) fixed in 4% buffered formalin for 10 minutes (BF group); (2) treated with 2% Carbowax in 50% ethanol (CW) for 10 minutes and then fixed in 100% ethanol for one hour (CW + ET); or (3) treated with CW for 10 minutes and then fixed in 4% buffered formalin for one hour (CW + BF). Cell morphology, adhesion to the glass slides and TUNEL reactivity were compared among the three groups. The effects of prolonged prefixation of cell suspensions in CW and of the postfixation of cell smears in BF for one, three and seven days were also examined. RESULTS CW + BF treatment yielded satisfactory cell morphology, minimum cell loss and an excellent TUNEL reaction. However, prolonged prefixation in CW resulted in cell shrinkage. CONCLUSION CW + BF treatment can be widely recommended for use with cytologic preparations for the TUNEL assay. |
| Databáze: | OpenAIRE |
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