Cryoembedding and sectioning of cochleas for immunocytochemistry and in situ hybridization.
Current emphasis on biochemical and molecular aspects of cochlear anatomy underscores the necessity for high quality cryostat sections of the inner ear. The large volume of fluid space within the cochlea makes cryoembedding and sectioning of the organ more problematic than that of other, more homogeneous tissues. Our method for cryoembedding of cochleas for immunocytochemistry and in situ hybridization uses slow infiltration with increasing concentrations of sucrose followed by degassed embedding medium before final orientation and freezing. This method permits high quality cryosections to be cut which preserve overall structure and cellular resolution.
Whitlon
D S
DS
Audiology and Hearing Sciences Program and Institute for Neuroscience, Northwestern University, Chicago, IL, USA. whitlon@northwestern.edu
Szakaly
R
R
Greiner
M A
MA
D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D006198:Hair Cells, Auditory / Q000737:chemistry / Q000166:cytology*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D008815:Mice, Inbred Strains; D008867:Microtomy / Q000379:methods*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics; D016610:Tissue Embedding / Q000379:methods*
D009419:Nerve Tissue Proteins