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Originally published as JHC exPRESS on January 5, 2009.
doi:10.1369/jhc.2008.952671
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Journal of Histochemistry and Cytochemistry
Volume 57 (8): 709-719, 2009
Copyright ©The Histochemical Society, Inc.

A Stereological Approach for Estimation of Cellular Immunogold Labeling and Its Spatial Distribution in Oriented Sections Using the Rotator

John Milton Lucocq and Christian Gawden-Bone

Division of Cell Biology and Immunology, School of Life Sciences, University of Dundee, Dundee, Scotland, United Kingdom

Correspondence to: Dr. John Lucocq, Division of Cell Biology and Immunology, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, Scotland, United Kingdom. E-mail: j.m.lucocq{at}dundee.ac.uk

Particulate gold labeling applied to ultrathin sections is a powerful approach for locating cellular proteins and lipids on thin sections of cellular structures and compartments. Effective quantitative methods now allow estimation of both density and distribution of gold labeling across aggregate organelles or compartment profiles. However, current methods generally use random sections of cells and tissues, and these do not readily present the information needed for spatial mapping of cellular quantities of gold label. Yet spatial mapping of gold particle labeling becomes important when cells are polarized or show internal organization or spatial shifts in protein/lipid localization. Here we have applied a stereological approach called the rotator to estimate cellular gold label and proportions of labeling over cellular compartments at specific locations related to a chosen cell axis or chosen cellular structures. This method could be used in cell biology for mapping cell components in studies of protein translocation, cell polarity, cell cycle stages, or component cell types in tissues. (J Histochem Cytochem 57:709–719, 2009)

Key Words: stereology • rotator • immuno-EM • gold labeling • quantification • polarity


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