CellTrace™ Far Red & CellTracker™ Deep Red
Long term monitoring of live cells in vitro or in vivo presents considerable challenges. Fluorescent tracers are used as one of the most common methods. However, fluorescent molecules usually are limited by cytotoxicity and/or lack of persistence within live cells over sufficient time periods. In this study, we evaluated our newly developed cell tracing/tracking reagents, CellTrac Far Red and CellTracker Deep Red, as novel red laser-excitable fluorescent dyes for flow cytometry and live cell imaging applications. These dyes cross the plasma membrane and covalently bind proteins inside cells, where the stable, well-retained fluorescent label offers a consistent, reliable fluorescent signal without affecting morphology or physiology and also exhibiting low cellular toxicity. CellTrace Far Red was demonstrated to be an excellent probe for cellular proliferation studies using flow cytometry, allowing measurement of up to eight generations of proliferating cells. CellTracker Deep Red was shown to be an excellent reagent for the study of cell movement, location and morphology by labeling and imaging live-cells with fluorescence microscopy. In addition, CellTracker Deep Red has excellent cell retention characteristics with bright fluorescence intensity even after 72 hours of cell staining, with signal retained after fixation, and can be multiplexed with other fluorescent dyes.
1. Coutu D.L., SchroederT. 2013. Probing cellular processes by long-term imaging-historic problems and current solutions. J. Cell Sci., 126, 1-11
2. Mattoussi H., Palui G., Na H.B. 2012. Luminescent quantum dots as platforms for probing in vitro and in vivo biological processes, Adv. Drug Delivery Rev. 64, 138-166
3. Taylor A., Wilson K.M., Murray P., Fernig D.G., Levy R. 2012. Long-term tracking of cells using inorganic nanoparticles as contrast agents: are we there yet? Chem. Soc. Rev., 41, 2707-2717
4. Johnson I., Spence M.T.Z., Eds. 2010. The Molecular Probes Handbook: A Guide to Fluorescent Probes and Labeling Technologies, 11th ed.: Life Technologies Corp.: New York
5. Lyons A.B. 1990. Divided we stand:Tracking cell proliferation with carboxyfluorescein diacetate succinimidyl ester. Immunol. and Cell Biol. 77, 509-515
6. Bercovoci N., Givan A.L., Waugh M.G., Fisher J.L., Vernel-Pauillac F., Ernstoff M.S., Abastado J.P., Wallace P.K. 2003. Multiparameter precursor analysis of T-cell responses to antigen. J. Immunol. Methods, 276, 5-17
7. Lyons A.B., Parish C.R. 1994. Determination of lymphocyte division by flow cytometry. J. Immunol. Methods, 171, 131-137
8. Weston S.A., Parish C.R. 1990. New fluorescent dyes for lymphocyte migration studies: Analysis by flow cytometry and fluorescence microscopy. J. Immunol. Methods, 133, 87-97
9. Quah B.J.C., Parish C.R. 2012. New and improved methods for measuring lymphocyte proliferation in vitro and in vivo using CFSE-like fluorescent dyes. J. Immunol. Methods, 379, 1-14
10. Wallace P., Tario Jr. J., Fisher J., Wallace S., Ernstoff M., Muirhead K. 2008, Tracking antigen-driven responses by flow cytometry:Monitoring proliferation by dye dilution. Cytometry Part A. 73A, 1019-1034
11. Hawkins E., Hommel M., Turner M., Battye F., Markham J., Hodgkin P. 2007 Measuring lymphocyte proliferation, survival and differentiation using CFSE time-series data - pp2057- 2067
12. Fuse S., Usherwood E. 2007. Simultaneous analysis of in vivo CD8+ T cell toxicity against multiple epitopes using multicolor flow cytometry. Immun. Invest. 36:829-45
13. Horan P.K., Melnicoff M.J., Jensen B.D., Slezak S.E. 1990. Fluorescent cell labeling for in vivo and in vitro cell tracking. Methods Cell. Biol. 33, 469-490
14. Filby A., Begum J., Jalal M., Day W. 2015. Appraising the suitability of succinimidyl and lipophilic fluorescent dyes to track proliferation in non-quiescent cells by dye dilution. Methods 82:29-37
15. Hawkins ED, Hommel M., Turner ML. 2007. Measuring lymphocyte proliferation, survival and differentiation using CFSE time-series data. Nat. Protoc. 2:2057-67