TY - JOUR
T1 - Use of cryostat sections from snap-frozen nervous tissue for combining stereological estimates with histological, cellular, or molecular analyses on adjacent sections
AU - Schmitz, Christoph
AU - Dafotakis, Manuel
AU - Heinsen, Helmut
AU - Mugrauer, Kristina
AU - Niesel, Alexandra
AU - Popken, Gregory J.
AU - Stephan, Michael
AU - Van De Berg, Wilma D.J.
AU - Von Hörsten, Stephan
AU - Korr, Hubert
PY - 2000/11/9
Y1 - 2000/11/9
N2 - 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.
AB - 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.
KW - Cryostat sections
KW - Immunohistochemistry
KW - Snap-frozen
KW - Stereology
UR - http://www.scopus.com/inward/record.url?scp=0033789886&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/S0891-0618(00)00075-2
DO - https://doi.org/10.1016/S0891-0618(00)00075-2
M3 - Article
C2 - 11074341
SN - 0891-0618
VL - 20
SP - 21
EP - 29
JO - Journal of Chemical Neuroanatomy
JF - Journal of Chemical Neuroanatomy
IS - 1
ER -