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I’m seeing uneven or non-uniform staining with CellBrite® Red, what should I do?

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CellBrite® dyes are ready-to-use solutions of the lipophilic carbocyanine dyes DiI, DiO etc. It is challenging to get uniform labeling with CellBrite® dyes due to their hydrophobicity, CellBrite® Red (#30023), which is DiD,  is particularly difficult as the hydrophobic dye tends to aggregate and give uneven staining. Warming the dye vial to ~50°C to dissolve any dye aggregates before staining could be tried. Also rather than adding the dye directly to the medium, we recommend diluting in it medium (1:200 dilution) first, and then adding the medium with the dye to the cells. Also, be aware that these dyes will be internalized if cells are cultured after staining.

Alternatively, for live cell staining, CellBrite® Steady or the reactive dyes CellBrite® Fix or MemBrite® Fix would be more suitable as they result in more even staining.


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