Hello,
My name is Kudirat, I am from University of Barcelona at Department of Analytical chemistry mainly in Chemometrics group.
At this moment we are trying to obtain cryosections of crustacean in this case Daphnia Magna which has not been sucessful. The anatomy of the Daphnia is destroyed while cryosectioning, possibly due to the resistance of the carapace.
1) This is what we have done.
Firstly, we orientate the sample with 100 % OCT and flash freeze with liquid nitrogen. After then we stored at -80 ºC and cut between the range of 15 - 20 um
2) What we have not done,
Fix the sample, this is just because we thought it can damage our sample since we want to acquire images with FT-IR spectroscopy.
We will appreciate any response which can help us.
Thank you
Yes I have sectioned Daphnia successfully but they were FFPE sections and not cryostat and not for FT-IR. You may well have a problem with the amount of water within the specimen. There may well be other problems due to the cryotomy technique.. You could try increasing strengths of a sucrose solution 10%, 20% 30% then OCT (let your specimen meld with the OCT for a short while) and snap freeze in dry ice chilled iso-pentane. Wrap it up for storage at -80C to stop it freeze drying. Good luck..
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