Dear All,
I cultured HUVECs in GelMA+Fibrin construct. But when I do the put DAPI in, it stained both HUVECs and Fibrin network. So the DAPI signal from Fibrin disturbs the signal from HUVECs, and it might lead to misinterpretation.
Is there anyway to eliminate/minimise the DAPI signal from fibrin?
Or subtract it, or even any strong argument to neglect this issue
Best Regards,
Yannapol S.
Hi Yannapol,
autofluorescence from polymers is quite often a problem (see: Article Pre-culture Sudan Black B treatment suppresses autofluoresce...
)As an alternative you might try to stain your (fixed) cells with propidiumiodide and hope that autofluorescence is less of a problem @ 555nm?
Best regards,
Christian
Wait so did the DAPi stain the proteins or is it just autofluorescence from them contaminating the signal? If the DAPIs stained them you think then you could try lowering the concentration and incubating for a longer period of time.
Otherwise you could also use a different nuclear dye. The TOTO and TOPRO series of dyes from invitrogen spans the whole spectral range and is often well out of the way of autofluorescent signals.
I would also suggest you to use a different dye with different spectral range would be helpful, like Howard suggested.
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