Dear Histonetters,
I have a person using acetic acid as a fixative for frozen sections containing bioluminescent proteins (GFP and tdTomato, a red fluorescent protein). I have always been taught to use acetic acid in combination with other chemical agents (e.g. as in Bouins or Carnoys) but not alone due to the swelling of cells and tissues and probably other acid protein hydrolysis going on too. Has anyone ever run across using acetic acid (concentration is not high, but not known by me)? She does encounter problems with GFP staining, but the tdTomato still glows although her results are inconsistent. I have no clue where this fixation method originated from but I suspect it may be from a fixative where something was lost in making it up/forgotten/left out - ending up with acetic acid alone. A quick look in Sheehan and Hrapchak's Theory and Practice of HIstotechnology, indicated acetic acid was rarely used alone for fixation. I am off to John Kiernan's book after sending this message. I understand the sensitivity of GFP and some other GFP chimeras , also RFP (closely related structurally to GFP) to alcohols and other organic solvents, heat, and pH where fluorescence is lost and/or diminished significantly. Enlighten me please Gayle M. Callis HTL/HT/MT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
