PMID-25855189 Mapping Synapses by Conjugate Light-Electron Array Tomography
- Use aligned interleaved immunofluorescence imaging follwed by array EM (FESEM). 70nm thick sections.
- Of IHC, tissue must be dehydrated & embedded in a resin.
- However, the dehydration disrupts cell membranes and ultrastructural details viewed via EM ...
- Hence, EM microscopy uses osmium tetroxide to cross-link the lipids.
- ... Yet that also disrupt / refolds the poteins, making IHC fail.
- Solution is to dehydrate & embed at cryo temp, -70C, where the lipids do not dissolve. They used Lowicryl HM-20.
- We show that cryoembedding provides markedly improved ultrastructure while still permitting multiplexed immunohistochemistry.
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