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. 2010;59(6):527-30.
doi: 10.1093/jmicro/dfq064. Epub 2010 Aug 23.

A novel method for large-scale immuno-SEM using protein G coupled polystyrene beads

In-Hwan Song  1 , James E Dennis
Affiliations

A novel method for large-scale immuno-SEM using protein G coupled polystyrene beads

In-Hwan Song et al. J Electron Microsc (Tokyo). 2010.

Abstract

Polystyrene beads with a mean diameter of 0.76 μm were coupled with protein G and then anti-type II collagen IgG or anti-chondroitin-4-sulphate IgG were tagged to protein G. Antibody-tagged beads were applied to articular cartilage and labelled beads were counted in each sample. Antibody-tagged beads labelled significantly higher than IgG isotype control. We propose immuno-SEM using protein G coupled beads as a valuable method for micrometre range observation for specific protein distribution on surfaces of tissues or organs. This will provide information about structure as well as antigenicity on the surface at the same time.

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Figures

Fig. 1
Fig. 1
Immunolabelled beads for type II collagen on native synovial surface and cut exposed surface of articular cartilage in rabbit femur condyle. Anti-type II collagen IgG-tagged beads labelled significantly higher than IgG untagged control. Asterisk indicates lacuna; scale bar = 5 μm.
Fig. 2
Fig. 2
Immunolabelled beads for chondroitin-4-sulphate on native synovial surface and cut exposed surface of articular cartilage in rabbit femur condyle. Anti-chondroitin-4-sulphate IgG-coated beads labelled significantly higher than IgG untagged control. Asterisk indicates lacuna; scale bar = 5 μm.
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