I am using 4% paraformaldehyde to fix whole blood at the final concentration of 2%. However, my fixation is for a very short while and I want to remove the paraformaldehyde from blood immediately. What kind of agents and to what concentration could be used to quench the residual paraformaldehyde? Glycine could be a solution, but does anybody have fair idea about the concentration, amount and time?
Dear Subhabrata,
the glycine use is a good idea. In my opinion, the molar glycine concentration should be equal to the molar concentration of the formaldehyde that has arisen from paraformaldehyde. While the formaldehyde concentration is approx. 0.67 M, the equimolar concentration of the glycine is approx. 5%. This concentration will be hypertonic because the molecule of the glycine does not permeate so free as the molecule of the formaldehyde into cells. Therefore I suppose after some time the blood cells isolate after cca 10 minutes by centrifugation. The reaction is better performing at lowered temperature and the glycine solution adds slowly under cooling.
This is theoretical conclusion. It is necessary to try it experimentally.
Best regards
Pavel
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