Immunocytochemistry of resin sections

This step relies on the silver enhancement of colloidal gold for detection. To keep background to a minimum, sections are washed in water containing a minimum of ions—which can act as nucleation sites for the silver—before and after the enhancement. 

Protocol for labelling resin sections for light microscopy 

Mark around the section with Dako (or similar) pen. 

Incubate with first antibody diluted in PBS/BSA overnight in moist chamber. 

Incubate with gold conjugate diluted in blocking buffer for 90 min. 

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Protocol for labelling resin sections for hght microscopy 260 Electron microscope immunocytochemistry of resin sections 261... 

A second approach to postembedding electron microscopic immunocytochemistry is the use of special aqueous resins, while these resins are still hydrophobic, they tolerate some aqueous material in the tissue. Examples of these resins are LR White or Lowicryl K4M. Another approach with aqueous embedding is to freeze tissue in buffer very rapidly, and to section with a cryo-ultramicrotome. Cryo-ultramicrotomy is a technique requiring considerable training and cannot be used by the novice. 

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