[Histonet] Wrinkles on Glutaraldehyde Fixed Tissue

[Ricardo Leyva]

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
rleyva <@t> ucsd.edu
UCSD Shiley Eye Center