Improved CFSE Alternatives for Cell Proliferation

On this page:

• Selection Guide for CellTrace Cell Proliferation Kits
• How CellTrace proliferation kits work
• Protocols
   

CellTrace Cell Proliferation Kits contain a cell-permeant nonfluorescent ester of an amine-reactive fluorescent molecule, which enters cells by diffusion through the plasma membrane. Upon entry into the cell, the nonfluorescent molecule is converted to a fluorescent derivative by cellular esterases. The active succinimidyl ester covalently binds to amine groups in proteins, resulting in long-term dye retention within the cell.

Through subsequent cell divisions, daughter cells receive approximately half of the fluorescent label of their parent cells, allowing the analysis of the fluorescence intensities of cells labeled and grown in vivo.

Analysis of the level of fluorescence in the cell populations by flow cytometry permits the determination of the number of generations through which a cell has progressed since the label was applied (Figure 1).

To understand more about the suitability of these reagents for tracking proliferation as compared to lipophilic fluorescent dyes, Filby and coworkers published a method to assess and compare the performance of three succinimidyl ester–based dyes (including Invitrogen CellTrace Violet and CellTrace Far Red dyes) and two lipophilic dyes.

Article: Appraising the Suitability of Succinimidyl and Lipophilic Fluorescent Dyes to Track Proliferation in Non-Quiescent Cells by Dye Dilution

The CellTrace stains have been detected after in vitro labeling for several days, or approximately 7–8 division cycles, before being overwhelmed by the natural autofluorescence of the cells. Up to 10 cell divisions have been observed using the CellTrace Violet Cell Proliferation Kit on a violet laser–equipped flow cytometer (Figure 2).

Article: A comparison study of CellTrace Violet, Yellow, and Far Red

The CellTrace Cell Proliferation Kits are available for the UV, violet, blue, yellow (561 nm), and red lasers (Figure 3), allowing researchers to analyze cell proliferation in combination with other live-cell applications (i.e., immunophenotyping, cell sorting, and cell-cycle analysis), maximizing the information that can be collected in a single experiment, with minimal compensation.

CellTrace Violet and CellTrace Far Red are also compatible with commonly used green-fluorescent dyes like FITC and Invitrogen Alexa Fluor 488, or with green fluorescent proteins (i.e., GFP) (Figure 4).

Poster: Multiplexing CellTrace Violet with common flow cytometry reagents
Poster: Multiplexing CellTrace Yellow with common flow cytometry reagents
Article: Multiplexing CellTrace Far Red with GFP