Hello,
We tried to do CLEM experiment on sapphire disk including cryo fixation (Leica ICE) and then cryo substitution using LR white resin.
Once the resin is polymerized, we can't remove the sapphire disk.
I have tried Thermic shock but this resin crack and fragment itself once in LN2.
I have tried also to glue the sapphire part on a glass slide but when I remove the resin , the disk is attached on the support but take also few Lr white which contains the cells.
Have you an idea to remove easily the sapphire?
Thank you in advance
If the light imaging allows for it you could try to deposit a thin carbon layer on the sapphire disks on the sides the cells are grown on, that could act as a separation layer between the block and the disk? At least that works well with Epon.
Thank you for your answer,
The sapphire is already carbon coated (with a pattern to recognize area of interest)
I can't make a whole carbon coating as I will lose this pattern.
But thank you for your advise
Just to clarify, why are you using LRW? Immuno labeling? Otherwise a Epon resin will be easier to work with.
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