Hello everyone,
I have fixed 22-week-old rat femurs in 10% formalin for 24hrs, then decalcified in 10% EDTA for 28 days.
I have read a lot recently that over-fixation is not too much of an issue compared to under-fixation.
Is it possible to take my bones out of EDTA and put them back into formalin for e.g. 24 more hours, prior to putting them in the autoprocessor?
What would be the optimal time to formalin fix whole femurs from rats of this age?
Thanks for reading,
Flynn
What would be the point? You've already fixated your samples (cross linked most proteins) so more fixation is not going to help or do anything. If you think the fixation process is not working properly you probably need to incubate your samples longer then 24hrs, a couple of days will do the trick. I have never tried this method myself but it looks interesting as an alternative to formalin. Article HistoChoice (R) as an alternative to formalin fixation of un...
Hi Aart Van Apeldoorn . I guess I was thinking that 48hrs total fixation time would be better than 24hrs?
Even if it was split 24hrs before and 24hrs after decalcification?
Yes probably much better, keep in mind that bone is quite a dense tissue. So you need to take your time to properly fix your samples. Fixating afterwards is not going to help you. Any harsh treatment you did will already have had a negative effect on the cells. The point of fixation is to "freeze" the samples in time. By fixation you stop all cellular processes and preserve the anatomy of cells in their current state. It's crucial to do this right, just take your time and you probably get much better histological outcomes.
I fixed rat knees for 3 days in 10%NBF. All procedures like Immuno and special stain procedures worked
- Badges
- Science topic
- Similar topics
- Microbiology
- Assays