A quantitative method for assessing co-localization in immunolabeled thin section electron micrographs

A method is introduced for the analysis of nearest neighbor distances between immunogold particles marking proteins on electron micrographs. Deviation from the distribution that is predicted by chance indicates co-localization of the labeled species, and the potential for productive interaction in vivo. Application of this method to the analysis of nearest neighbor distances in experiments with pea leaf thin sections and isozyme-directed antibodies indicates that glyceraldehyde-3-P dehydrogenase is located near P-glycerate kinase and near aldolase in the chloroplast stroma, consistent with the notion that these enzymes are part of a multi-enzyme photosynthetic CO2-fixation complex in situ. (C) 2003 Elsevier Inc. All rights reserved.