Has anyone used a glyoxal based solution (pH=4 as described in: Article Glyoxal as an alternative fixative to formaldehyde in immuno...
) for transcardial perfusion in mice. I see that in the original paper they only use it for the fixation of regions that were dissected from unfixed animals and then fixed by immersion. I was wondering about undesirable effects of transcardially perfusing a pH=4, ethanol containing solution... Any advice or experience on this?Cheers,
J
Hi J. Ramirez-Franco , did you test this yet? If you look in the supplementary content you can see that one set of experiments did involve transcardial perfusion:
"Mice were perfused with 12 ml 0.9% NaCl solution, and then with either 24 ml 4% paraformaldehyde (PFA) in Phosphate Buffer Saline (PBS) or 24 ml 3% pH 4 glyoxal fixative solution, both freshly made just prior to use. "
I tried this last week and I'm not sure how to judge the result. While there was clearly fixation it looked pretty terrible (beaded staining, some stainings failed 100%). I perfused with 20ish ml of ph ~5 glyoxal fixative solution, and post-fixed in the same solution overnight at 4C. I'll give it one more shot, but in the meantime, have you had any success?
Best,
Sverre