I have used FAA fixative (10% Formalin, 5% Acetic Acid, 50% Ethanol) to fix tomato apices for in situ but after fixation and 30 min vacuum infiltration the tissue becomes red-brownish. Do you know what could be the reason for it?
Ethanol concentration may be very high (50%). You can minimize ethanol concentration
30 min is long for the vacuum infiltration, this could damage the plant tissue. A few minutes should be sufficient.
Thanks for the answers, I tried 4% PFA and 2% Triton and it worked better. Anyway I'll try to reduce the ethanol concentration. What would you recommend Omaima? 30%? Have you had that experience before and reducing ethanol helped you?
I have tried less vacuum, but that does not seem to be the problem. The brownish starts almost 3-5 minutes after dissections. Lately, I've being dissecting the tissue submerged into the fixative under the hood and that's been helping a bit.
Hello Suresh, for which reagent you mean high concentration?
Thanks to all!
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