High resolution image of early endosomes in which the membranes are easily seen as a tripartite track. Gold, labeling the C6 antigen, is observed only in the body of the early endosome and not in the coated evagination or small coated vesicle lying nearby, which may be a newly released carrier vesicle or a coated evagination still attached to an early endosome. TEM taken on 5/14/91 by R. Allen with Zeiss 10A operating at 80kV. Neg. 31,500X. Cells were lightly fixed with 0.25% glutaraldehyde and infiltrated with 2.3M sucrose before being frozen in liquid nitrogen and thin sectioned at a temperature of –100?C at approximately 75nm thickness. Frozen sections from these preparations were then thawed, washed, and exposed to a monoclonal primary antibody that was raised in mice or rabbit/goat and to colloidal gold-complexed goat-anti-mouse/rabbit secondary antibodies. Further details of preparation are outlined in Methods Cell Biol. 2010;96:143-73. The raw film was scanned with a Epson Perfection V750 Pro. This image is best used for quantitative analysis. Additional information available at (http://www5.pbrc.hawaii.edu/allen/).
Biological Process: Endosome organization
Author: Richard Allen (University of Hawaii)
Source: The Cell: An Image Library