I fixed bacterial cells in 2.5% glutaraldehyde and dehydrated them in a series of ethanol solutions, ending with 100% ethanol. I intended to proceed with critical point drying (CPD), but the machine was unavailable. As an alternative, I air-dried the cells and stored them in a desiccator for 3 days. Now I want to take it back to CPD, my sample is still okay to do?
Thank you in advance
Hello,
Your sample is dry, no need to use the CPD.
Additionally you will put it again in liquid CO2 (directly or passing by solvent - acetone?) to dry it again.
If air-drying gives a poor structure I don't think CPD will ameliorate it.
So useless additional work.
Ah! And you could have keep your sample in ethanol while the CPD was busy.
Nicolas Barois Thank you for your suggestion. I recently read on Researchgate that storing bacteria in 100% ethanol for an extended period can damage their cellular structure. Additionally, air-drying them may also be detrimental. I'm unsure about the best method for preserving these bacteria at that time.
Nicolas Barois I apologize for bothering you again. I’m having an issue. I brought my bacterial samples back to the CPD today, but the technician said the machine wasn’t working, so the samples didn’t dry after CPD. They’ve been put back in 100% ethanol again twice now. I don't have liquid Co2 and HMDS. I'm not sure how to do it. Do you have any ideas? Can I rehydrate back to 50%?
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