Establishment of Flow-FISH method for simultaneous detection of telomere length and cell differentiation antigen
10.3760/cma.j.issn.0254-5101.2010.03.019
- VernacularTitle:Flow-FISH同时检测细胞端粒长度及表面分化抗原方法的建立与优化
- Author:
Yu ZHAN
;
Ru FENG
;
Zhengshan YI
;
Lanlin SONG
- Publication Type:Journal Article
- Keywords:
How-FISH;
Telomere length;
Differentiation antigen;
Leukemia
- From:
Chinese Journal of Microbiology and Immunology
2010;30(3):267-272
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the Flow-FISH method for simultaneous detection of telornere length and cell differentiation antigen. Methods HL60, Raji, Molt4 cells were cultivated. Each step and the conditions of the Flow-FISH procedure were optimized, standardized and validated, then 14 acute leukemia patients were observed for the changes of telomere length combined with differentiation antigen after complete remission by the method. Results Cells were stained with Alexa Fluor(R) 647-labeled antibody. Anti-gen-anfibedy complexes were covalenfly cross-linked onto the cell membrane before telomere staining. Cells were hybridized with telomere-specific fluorescein isothiocyanate (FITC)-conjugated peptide nucleic acid (PNA) probes followed by being counterstained with propidium iodide(PI). Multicolor Flow-FISH was performed to analyze telomere length and differentiation antigen simultaneously. The patients showed longer telomere and lower antigen expression after complete remission. Conclusion The Flow-FISH method for simultaneous detection of telomere length and differentiation antigen was successfully established which might prove to be a promising means for leukemia research especially in those without special molecular markers.
