I want to do a cell migration study for around 4 hours and need to fluorescently label the cells with a dye. What do you recommend?
Calcein, AM and FDA (fluorescein diaceate) are examples of some dyes used for this application. Since these dyes are not incorporated or covalently attached to any cellular components, they may have a short retention time as some cell types may actively efflux the dye out of the cells. The CellTracker and CellTrace dyes include either a mild thiol-reactive chloromethyl group or amine-reactive succinnimidyl ester group to allow for covalent binding to cellular components, providing for better retention. As with any reagent, one should empirically determine retention times for the cell type used.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
I labeled my cells with Calcein, AM, but when I imaged the next day, there was no fluorescence from Calcein. Why?
Calcein, AM is a good choice for cell tracking and as a general cytoplasmic stain. However, it doesn't bind to anything and may be actively pumped out of the cells within a couple hours, which is likely what happened. The retention of Calcein within live cells is dependent upon the inherent properties of the cell type and culture conditions.
For long-term imaging, you may wish to consider a reactive cytoplasmic stains such as CFDA, SE or the CellTracker and CellTrace dyes.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
What is the shelf life of CellTrace CFSE Cell Proliferation Kit (Cat. No. C34554, C34570)?
CellTrace CFSE Cell Proliferation Kit is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended (in freezer (-5 to -30 degrees C) and protected from light). Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf).
For staining with CellTrace CFSE Cell Proliferation Kit, for flow cytometry (Cat. No. C34554 or C34570), up to how many cells can I have in 1 mL volume?
The cell concentration can be up to 1 x 10E7 cells/mL.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
I would like to label two cell populations with two different CellTrace reagents and then co-culture these cells. Will the CellTrace reagent leave the cells to stain other cells?
We have not tested the use of the CellTrace reagents for co-culture applications. In theory, this may work, but you would have to test this on your cells of interest.
Find additional tips, troubleshooting help, and resources within our Cell Tracing and Tracking Support Center.