Cryosectioning fixed and cryoprotected biological material for immunocytochemistry.
Immunocytochemistry for transmission electron microscopy provides important information on the location and relative abundance of proteins inside cells. Gaining access to this information without extracting or disrupting the location of target proteins requires specialized preparation methods. Sectioning frozen blocks of chemically fixed and cryoprotected biological material is one method for obtaining immunocytochemical data. Once the cells or tissues are cut, the thawed cryosections can be labeled with specific antibodies and colloidal gold probes. They are then embedded in a thin film of plastic containing a contrasting agent. Subcellular morphology can be correlated with specific affinity labeling by examination in the transmission electron microscope. Modern technical advancements both in preparation protocols and equipment design make cryosectioning a routine and rapid approach for immunocytochemistry that may provide increased sensitivity for some antibodies.
Ahmanson Advanced Electron Microscopy and Imaging Center, House Ear Institute, Los Angeles, CA, USA.
Paul
P
Webster
Alexandre
A
Webster
D000818:Animals; D002417:Cattle; D016616:Cryoultramicrotomy / Q000295:instrumentation / Q000379:methods*; D003713:Dendritic Cells / Q000378:metabolism / Q000648:ultrastructure; D005404:Fixatives; D005976:Glutaral; D007150:Immunohistochemistry / Q000379:methods*; D007515:Islets of Langerhans / Q000648:ultrastructure; D051379:Mice; D016253:Microscopy, Immunoelectron / Q000379:methods*; D009297:Nasal Mucosa / Q000378:metabolism / Q000648:ultrastructure; D016707:Tissue Fixation
D005404:Fixatives; D005976:Glutaral