Immunogold TEM

Immunogold TEM is a technique used to examine the localization of proteins/molecules of interest labelled with colloidal gold-conjugated antibodies.

Equipment:

Hitachi H7600 TEM
FEI Tecnai G2 Twin TEM
Leica EM IGL Immunogold labelling system

Sample preparation:

1. Fixation: Cryo-fixation or Chemical fixation using aldehyde fixatives
2. Dehydration
3. Infiltration/embedding in Hydrophilic LR White or HM20 resin
4. Ultramicrotome sectioning
5. Antibody incubation for Immunolabelling
6. Grid Staining with uranyl acetate and lead citrate
7. TEM imaging