We are using fast red substrate that can be seen suing both chromogenic and fluorescence images. Until now we have used only hematoxylin as counterstaiing, but we are wondering if both hematoxylin and Dapi can be used to counter-stain the same section.
It is likely that hematoxylin will quench DAPI fluorescence. Hematoxylin has a broad absorption spectrum and it largely overlaps with DAPI emission. The attached image shows the normalized emission of DAPI overlapped with the absorption of Hematoxylin. It is generally done with serial sections.
I totally agree with Uttam. If you would like to try a different nuclear stain for brightfield applications I would suggest using methyl green on its very traditional way instead. You can find the absorption spectrum in the linked paper. It looks like it may suit to your purposes, as it would not absorb DAPI's fluorescence and probably nor fast red's (in principle), without interfering with your brightfield stuff.
Please let me know!
best of luck,
Daniel
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