Fura Red™，AM，细胞通透性

This is a known drawback of Calcium Crimson, AM (as well as Calcium Orange, AM and Fura Red, AM, which are also in the same emission range). You can try increasing the concentration and washing out of any unlabeled dye from the media, to try to get better signal-to-background. If that fails, we recommend using Rhod-3 AM instead, which has a much better change in signal in that wavelength.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Unfortunately, we do not have a user manual for the Fura Red, AM, cell permeant (Cat. No. F3021). You can reference section 19.3 of the Molecular Probes handbook for recommendations for use.
There is also a large number of citations & references provided on the product page.

Basically, you can dissolve the product in good quality anhydrous DMSO to prepare a stock solution of 1-10 mM and use a final solution of about 1-10 µM in a physiological buffer. You can then incubate for 15 to 60 minutes, wash and then leave the cells for about 30 minutes to allow the AM group to hydrolyse before starting the assay. You can add Pluronic F-127 (e.g., Cat. No. P6866) to facilitate uptake. Store the DMSO stock solution at -20 degrees C.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.