Morphometric analysis of gold particles in low-label cell compartments.
Quantitative estimation of the binding of gold-conjugated ligands to various cell organelles has become a commonly used method to quantify the amount of ligand-binding sites associated with those organelles. However, often a small percentage of organelles is labeled or the density of gold labeling is low. We have defined the "gold-labeled region" as a zone that has a boundary defined by the localization of the outermost gold particles. Such a phenomenon was recently observed in a study of the internalization of gold-labeled native surfactant into lamellar bodies of cultured pulmonary type II cells. We estimated the size and density of gold-labeled regions in lamelar bodies using a simple stereological approach and demonstrated that the low percentage of gold-labeled organelles can be explained as a result of the probability of random selecting through the labeled areas. Our method, which permits use of transmission electron microscopy to calculate the true parameters of gold-labeled regions, can significantly facilitate analyses of ligand binding to various cell compartments.
Department of Pathology, Sackler School of Medicine, Tel-Aviv University, Ramat-Aviv, Israel.
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Hammel
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Kalina
D000818:Animals; D006046:Gold; D008297:Male; D008854:Microscopy, Electron; D015388:Organelles / Q000648:ultrastructure*; D011650:Pulmonary Alveoli / Q000648:ultrastructure*; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling
D006046:Gold