Multiplexing analysis of cell proliferation and cellular functions using a new multicolor panel of fluorescent cell proliferation dyes (P1290)

2013 
Flow cytometry combined with fluorescent staining is a powerful tool for monitoring viable cells. CFSE is the most popular fluorescent dye for cell proliferation studies. However, the identical Ex/Em spectra to GFP or FITC make CFSE impossible to use for GFP cells or the applications where a FITC-labeled antibody is used. The other disadvantages of CFSE include the high background fluorescence and severe fluorescence intensity decrease on the 2nd generation of cells. We report the functional analysis of cell proliferation using a new class of multicolor CytoTell™ dyes. CytoTell™ dyes are functionally similar to CFSE since they contain both an esterase-cleavable and a cell-retaining group. They are well excited at major laser lines like 405, 488 or 633nm with multicolor emissions. We demonstrated that Cytotell™ dyes have a few significant advantages over CFSE. 1) They have minimal cytotoxicity and well retained in cells since they do not indiscriminately react with proteins, and have minimal MDR interaction; 2) They exhibit much faster response and are more convenient to use than CFSE. There is no fluorescence intensity gap between 1st and 2nd generation of cells. 3) There is no need to remove medium since CytoTell™ dyes do not react with medium components; 4) They are much more stable than CFSE. The stock solutions can be stored at RT for a few days; and (5). CytoTell dyes can be readily used for the multicolor applications with either GFP cells or FITC-labeled antibodies.
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