Quantifying immunogold labelling patterns of cellular compartments when they comprise mixtures of membranes (surface-occupying) and organelles (volume-occupying)

Histochemistry and Cell Biology
T M Mayhew, John M Lucocq

Abstract

In quantitative immunoelectron microscopy, subcellular compartments that are preferentially labelled with colloidal gold particles can be identified by estimating labelling densities (LDs) and relative labelling indices (RLIs). Hitherto, this approach has been limited to compartments which are either surface occupying (membranes) or volume occupying (organelles) but not a mixture of both (membranes and organelles). However, some antigens are known to translocate between membrane and organelle compartments and the problem then arises of expressing gold particle LDs in a consistent manner (e.g., as number per compartment profile area). Here, we present one possible solution to tackle this problem. With this method, each membrane is treated as a volume-occupying compartment and this is achieved by creating an acceptance zone at a fixed distance on each side of membrane images. Gold signal intensity is then expressed as an LD within the membrane profile area so created and this LD can be compared to LDs found in volume-occupying compartments. Acceptance zone width is determined largely by the expected dispersion of gold labelling. In some cases, the zone can be applied to all visible membrane images but there is a potential problem...Continue Reading

References

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Citations

Jun 17, 2008·Histochemistry and Cell Biology·Terry M Mayhew, John M Lucocq
Dec 1, 2009·Histochemistry and Cell Biology·Wolfgang-Moritz Heupel, Detlev Drenckhahn
Feb 5, 2010·American Journal of Respiratory and Critical Care Medicine·Connie C W HsiaUNKNOWN ATS/ERS Joint Task Force on Quantitative Assessment of Lung Structure
Jan 16, 2009·Cardiovascular Pathology : the Official Journal of the Society for Cardiovascular Pathology·Christian MühlfeldTerry M Mayhew
Jan 13, 2009·Annals of Anatomy = Anatomischer Anzeiger : Official Organ of the Anatomische Gesellschaft·Terry M MayhewMatthias Ochs
Jan 1, 2016·American Journal of Physiology. Lung Cellular and Molecular Physiology·Saket AhujaPoornima Mahavadi

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