Abstract
The dynamics of lipoprotein binding and endosome formation on pigeon monocyte-derived macrophages have been studied using Allen video-enhanced contrast, differential interference-contrast microscopy (AVEC-DIC) and nanovid (video-enhanced brightfield) microscopy. Subsequent to video-microscopy the ultrastructural (3-D) organization was examined by correlative Intermediate Voltage Electron Microscopy (IVEM). The use of gold “finder” grids mounted on glass coverslips provided the mechanism by which cells after being observed in the video system were located in the electron microscope. The lipoprotein, beta migrating low density lipoprotein (βVLDL), was conjugated to gold colloids (15-40 nm) to enhance visibility in both video-microscopy and IVEM. Subsequent to video microscopy, the cells were fixed with glutaraldehyde, dehydrated through ethanol and dried from CO2 by the critical point method for whole-mount EM. Such a correlative approach allowed static ultrastructural features to be associated with cellular dynamics as observed and recorded by video.
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