期刊
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
卷 62, 期 4, 页码 265-275出版社
SAGE PUBLICATIONS LTD
DOI: 10.1369/0022155413520404
关键词
TMRE; mitochondrial dye; caspase; cell sorting; Click-IT; viability; apoptotic cell elimination
类别
资金
- NIH [S10 RR023459]
- Harvard Pilot grant
- Russian Foundation for Basic Research [11-04-01749a, 13-04-40189-H]
- Program in Cellular and Molecular Medicine, Boston Children's Hospital
High-speed fluorescence-activated cell sorting is relevant for a plethora of applications, such as PCR-based techniques, microarrays, cloning, and propagation of selected cell populations. We suggest a simple cell-sorting technique to eliminate early and late apoptotic and necrotic cells, with good signal-to-noise ratio and a high-purity yield. The mitochondrial potential dye, TMRE (tetramethylrhodamine ethyl ester perchlorate), was used to separate viable and non-apoptotic cells from the cell sorting samples. TMRE staining is reversible and does not affect cell proliferation and viability. Sorted TMRE+ cells contained a negligible percentage of apoptotic and damaged cells and had a higher proliferative potential as compared with their counterpart cells, sorted on the basis of staining with DNA viability dye. This novel sorting technique using TMRE does not interfere with subsequent functional assays and is a method of choice for the enrichment of functionally active, unbiased cell populations.
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