title: A low-cost technique to cryo-protect and freeze rodent brains, precisely aligned to stereotaxic coordinates for whole-brain cryosectioning.
abstract: A major challenge in the histological sectioning of brain tissue is achieving accurate alignment in the standard coronal, horizontal, or sagittal planes. Correct alignment is desirable for ease of subsequent analysis and is a prerequisite for computational registration and algorithm-based quantification of experimental data. We have developed a simple and low-cost technique for whole-brain cryosectioning of rodent brains that reliably results in a precise alignment of stereotactic coordinates. The system utilises a 3-D printed model of a mouse brain to create a tailored cavity that is used to align and support the brain during freezing. The alignment of the frozen block is achieved in relation to the fixed edge of the mold. The system also allows for two brains to be frozen and sectioned simultaneously. System components, procedural steps, and examples of the end results are presented.
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA. vpinskiy@gmail.com
authors.forename: Vadim
authors.initials: V
authors.lastname: Pinskiy
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Tolpygo
authors.forename: Jamie
authors.initials: J
authors.lastname: Jones
authors.forename: Kevin
authors.initials: K
authors.lastname: Weber
authors.forename: Neil
authors.initials: N
authors.lastname: Franciotti
authors.forename: Partha P
authors.initials: PP
authors.lastname: Mitra
mesh_terms: D000818:Animals; D001921:Brain; D015925:Cryopreservation / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D016616:Cryoultramicrotomy / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D013238:Stereotaxic Techniques / Q000191:economics / Q000295:instrumentation / Q000592:standards
keywords: 3-D registration and reconstruction; High-throughput processing; Histology; Tissue freezing and sectioning
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