As part of my medical doctoral thesis, I am trying to improve the cryopreservation process in our laboratory. To do this, I would like to compare different glycol-based freezing media. These are used for the cryopreservation of heart tissue at -80°C and should provide good results in cryosectioning and microscopy. The following four are on the shortlist:
“Tissue-Tek O.C.T. Compound” (Sakura Finetek)
“Cryomatrix” (Thermo Fisher Scientific)
“Neg-50 Frozen Section Medium” (Richard-Allan Scientific)
“Cryo-Gel” (Electron Microscopy Sciense)
Since the possibilities for research are mostly limited to the manufacturer's specifications, I would be very pleased to receive assessments, experience reports and further information.
Dear Matthias Schippers ,
I have checked the MSDS and do not see any big differences in the composition of the freezing media. Tissue-Tek OCT seemed to me not to be buffered (in contrast to Cryomatrix and Neg-50) For Cryo-Gel I do not find (in my quick search) a meaningful MSDS with the composition.
In my opinion/experience the correct way of freezing might be as (or even more) important as the freezing medium. We use 2-Methylbutan/Isopentan in a old can. Please find here some links which should giude you in the right direction:
https://www.youtube.com/watch?v=0CzGDSKMCJg
https://www.youtube.com/watch?v=n3SbudEegZE (might be the best one)
https://www.youtube.com/watch?v=KdfqH4zoy5M
Viel Erfolg
Sönke
There is a big difference between Cryoprotectant and Freezing Matrix Media. Cryoprotectant will prevent the tissues from being destroyed or having significant artifacts. Matrix media provide structural support while cutting in the cryostat. Your question is really, how do you protect your sample tissues prior to freezing? As stated above @Sonke Weinert there is a methodology to getting the sample tissue to -80C, and there are several methods of protecting tissues such as Sucrose or Gycerine:Glycol solutions.
Placing the tissues in Freezing Mediums (OCT etc) doesn't protect the internal cellular structures from freezer artifacts, mainly rupture.
That being said the second comparator of which freezing medium to use during sectioning is also of interest, but your optimization may vary when using different blades (high vs low profile) or of different manufacturers etc. The temperature of the Cryostat is also of importance as well as the temperature of the tissue and medium. There are many variables to account for in this project, it is not a "this is the best solution" for all instances.
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