Use of cryostat sections from snap-frozen nervous tissue for combining stereological estimates with histological, cellular, or molecular analyses on adjacent sections

Christoph Schmitz*, Manuel Dafotakis, Helmut Heinsen, Kristina Mugrauer, Alexandra Niesel, Gregory J. Popken, Michael Stephan, Wilma D.J. Van De Berg, Stephan Von Hörsten, Hubert Korr

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Adequate tissue preparation is essential for both modern stereological and immunohistochemical investigations. However, combining these methodologies in a single study presents a number of obstacles pertaining to optimal histological preparation. Tissue shrinkage and loss of nuclei/nucleoli from the unprotected section surfaces of unembedded tissue used for immunohistochemistry may be problematic with regard to adequate stereological design. In this study, frozen cryostat sections from hippocampal and cerebellar regions of two rat strains and cerebellar and cerebral regions from a human brain were analyzed to determine the potential impact of these factors on estimates of neuron number obtained using the optical disector. Neuronal nuclei and nucleoli were clearly present in thin sections of snap-frozen rat (3 μm) and human (6 μm) tissue, indicating that neuronal nuclei/nucleoli are not unavoidably lost from unprotected section surfaces of unembedded tissue. In order to quantify the potential impact of any nuclear loss, optical fractionator estimates of rat hippocampal pyramidal cells in areas CA1-3 and cerebellar granule and Purkinje cells were made using minimal (1 μm) upper guard zones. Estimates did not differ from data reported previously in the literature. This data indicates that cryostat sections of snap-frozen nervous tissue may successfully be used for estimating total neuronal numbers using optical disectors. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)21-29
Number of pages9
JournalJournal of Chemical Neuroanatomy
Volume20
Issue number1
DOIs
Publication statusPublished - 9 Nov 2000

Cite this