Home Knowledge Base After incubation with the dual detection reagent at 37° C for 30 min, the cells were not healthy and rounding up. Some cells were also detached from the plate. No specific staining on positive control was observed vs. DMSO controls.

After incubation with the dual detection reagent at 37° C for 30 min, the cells were not healthy and rounding up. Some cells were also detached from the plate. No specific staining on positive control was observed vs. DMSO controls.

Date Published: July 18, 2023

For fragile cells, the following modification to the standard protocol are suggested:

Use PBS/5% FBS as an assay buffer or complete medium, preferably without phenol red.
If cells still appear unhealthy, then decrease the time of the staining to 15-20 min.
Reduce the dye concentration to 1000 x dilution. This may require a longer exposure time for detection by microscopy.
The signal may be obscured due to photo bleaching. It is recommended to use mounting medium that prevents photo bleaching.

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After incubation with the dual detection reagent at 37° C for 30 min, the cells were not healthy and rounding up. Some cells were also detached from the plate. No specific staining on positive control was observed vs. DMSO controls.

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