Dear Colleagues, (This request is the second part following the first one)
As I mentioned before,our frozen sections are of poor quality.And I am reading a manual of "HistoGene LCM Frozen Section Staining Kit". I take great interest in preparing frozen sections which would have good quality and could be used to do LCM as well.
In the manual it says in the part of "Specimen Freezing",
1. Place cryomold on dry ice and fill it halfway with OCT.
2. Collect dissected tissue specimen.
3. Place tissue specimen in cold cryomold from step 1.
4. Add OCT to cold cryomold until specimen is covered.
5. Wait for OCT to turn white.
6. Store frozen specimen in the cryomold in a -70#161##227#C freezer.
And I think our poor quality may due to, We do not use "quick-freezing" technique.When we obtained specimen from bedside,we put it in a 4C container and take back to the lab.Then immerse the specimen in OCT.Then put them all in -29C in a cyrostat.Then sectioning...
So I just wonder if could I do as follow, fix the specimens firstly in a kind of fixative solution then transfer them to the lab at 4C .And then incubate it in 20% sucrose buffer for 30 minutes and immerse the specimen in OCT.Then put the OCT-specimen in -70C fridge in order to be quick freezed.Finally do cyrosectioning.
Would the idea I thought of above necessary and approachable? Would you kindly give me some advice? (BTW,solid CO2 and liquid nitrogen are all convenient for our technician in routine work so I want to try prefix and incubate in -70C fridge and so on)
Any comments are welcome. Thank you very much!
yichao Wu,Ph.D candidate