Currently I am trying to make multiple immunofluorescence labelling of formalin-fixed paraffin-embedded (FFPE) liver tissue. Unfortunately, tons of red blood cells with strong autofluorescence at 488 and 546 excitation were found in tissue sections, though I have made blocking procedure using 5% BSA before antibody staining.
Does anyone have any suggestions to reduce or eliminate autofluorescence by red blood cells? May cryosections help? Thank you very much in advance!
I agree with David that perfusion fixation will remove blood and after perfusion fixation you need to immerse your tissue again in the same perfusion fixative for 24 hours or less according to the tissue size.
Human liver samples, Can I use perfusion fixation method to remove blood cell as well?
Jinqing, are you treating biopsies? Autoptic tissues? Let me know.
Thank you Dr. Serafini! Our specimens were from the patients with HCC who received surgical resection before routine fixation. Are there any differences between biopsies and autopsies?
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