Why do you use glutaraldehyde for eyes? Davidson's is a much better fixative for eyes. With Davidson's you have two fast penetrating ingredients and one slow acting fixative. Glut is only a fast fixative. Besides, the best institutions who deal with eyes only use Davidson's. Just my thoughts.
Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 -----Original Message----- From: [email protected] [mailto:[email protected]] On Behalf Of Ricardo Leyva Sent: Tuesday, May 06, 2014 11:38 AM To: [email protected] Subject: [Histonet] Wrinkles on Glutaraldehyde Fixed Tissue Dear Histonetter, I am having an issue getting rid of wrinkles on sections from 4% glutaraldehyde fixed paraffin embedded ocular tissue. The laboratory's tissue of interest is the retina, and I use glut to avoid retinal detachment (this is mainly on tissue that won't be immunostained). I normally section at 5µm, and fix the tissue 1 or 2 days. I set the water bath at a temperature that is 5-10 C below the wax melting point. I have tried adding ethyl alcohol to the water and the wrinkles remain-or takes very long for most of them to be gone. I have tried placing the sections in a 1:15 mix of ethanol and water-and have also tried 30% ethanol in water, both at RT, before placing the sections on the water bath; when I do this, the sections separate abruptly (once in the water bath) from the slide I use for transferring the tissue, causing tissue layer separation. When I work with 10% formalin, the section wrinkles disappear nicely in the water bath that contains water and a bit ethanol. Nonetheless, when it comes to 4% glut it just does not work. Any input in troubleshooting this is greatly appreciated. Best, Ricardo Leyva (858) 822-1629 [email protected] UCSD Shiley Eye Center _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
