Cryosectioning and immunolabeling

Nat Protoc. 2007;2(10):2480-91. doi: 10.1038/nprot.2007.365.


In this protocol, we describe cryoimmunolabeling methods for the subcellular localization of proteins and certain lipids. The methods start with chemical fixation of cells and tissue in formaldehyde (FA) and/or glutaraldehyde (GA), sometimes supplemented with acrolein. Cell and tissue blocks are then immersed in 2.3 M sucrose before freezing in liquid nitrogen. Thin cryosections, cut in an ultracryotome, can be single- or multiple immunolabeled with differently sized gold particles, contrasted and viewed in an electron microscope. Semi-thin cryosections can be used for immunofluorescence microscopy. We describe the detailed procedures that have been developed and tested in practice in our laboratory during the past decades.

MeSH terms

  • Animals
  • Cells, Cultured
  • Cryoultramicrotomy / methods*
  • Humans
  • Immunohistochemistry / methods*
  • Lipids / analysis
  • Proteins / analysis
  • Rats
  • Tissue Fixation


  • Lipids
  • Proteins