[3dem] resin misbehaving at retina pigment granules

[Anna Pielach]

Hello all

I wonder if anyone here has experienced issues with resin infiltration/polymerization in the pigmented epithelium in mouse retina, the pigment granules to be more specific. We have been really struggling with this. The samples are fixed with Karnovsky fix (2% FA, 2.5 GA in cacodylate buffer). We have tried all sorts of combinations including microwave-assisted dehydration and resin infiltration in the Leica processor, switching to propylene oxide for 3 extra final solvent changes, Hard-Plus, Durcupan and LR White resins. The resin is consistently separating from the periphery of pigment granules and in many places this adds up to severe cracking of the sections. Please do let me know if you have managed to optimise something similar or have any suggestions.

All the best,
Anna

TEM:
[cid:image002.jpg at 01D756CC.C4ACE020]
[cid:image006.jpg at 01D756CC.C4ACE020]

SEM:
[cid:image007.jpg at 01D756CC.C4ACE020]
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