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Cellular Analysis
Fluorescence drop upon compound addition
Cells may become dislodged during addition. Decrease the rate of addition or seed fewer cells per well.
Tue, Jul 18, 2023
Chromatin condensation is not observed
Incorrect drug concentration or incubation time for cell type: Make sure the cell preparation protocol generates single cells with minimal…
Tue, Jul 18, 2023
Control cells without treatment show low orange signal.
Control cells are not healthy. Extended storage of cells after staining may adversely affect their health >> Use healthy cells…
Tue, Jul 18, 2023
High baseline fluorescence
Growth medium and organic anion transport contribute to high baseline fluorescence. Remove the medium before adding indicator dye to the wells.…
Tue, Jul 18, 2023
Healthy cells show very high fluorescence intensity
PMT setting is too high. Lower PMT setting on flow cytometer. Incubation temperature. Incubation with the dye should be at…
Tue, Jul 18, 2023
Poor Mito-ID staining observed.
Stained cells may have been exposed to strong light. Mito-ID® MP dye is light sensitive; be sure to protect samples…
Tue, Jul 18, 2023
Cells do not exhibit fluorescence after incubation with the detection reagent.
Cell viability is low. Cells should be in log growth phase. Cell samples cannot be kept longer than 6 hours…
Tue, Jul 18, 2023
Fluorescence does not increase after incubation with inhibitor.
Cells do not express any MDR1, MRP1/2 or BCRP. Use a positive control cell type expressing corresponding ABC transporter.
Tue, Jul 18, 2023
Inconsistent fluorescence shift using the same cell (irreproducible results).
Inadequate incubation condition. Always use a water bath (not incubator) and ensure temperature of water bath is 37°C.
Tue, Jul 18, 2023
Untreated cells have calcium response
Inconsistent DMSO concentration. Ensure that buffer used for the negative control wells has the same final concentration of DMSO as…
Tue, Jul 18, 2023