This micrograph shows the pegs on the surface of the decorated tubules associated with the contractile vacuole that are negatively stained by the uranyl acetate dissolved in the methylcellulose solution used to give support to the frozen then thawed section. This section was labeled by a polyclonal antibody to the V-ATPase of Dictyostelium which cross-reacts with the V-ATPase in Paramecium. 108:3163-3170, 1995). TEM taken on 2/28/92 by R. Allen with Zeiss 10A operating at 80kV. Neg. 31,500X. Bar = 0.2µm. Adapted with permission from J. Cell Sci. 108:3163-3170, 1995. 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 negative was printed to paper and the image was scanned to Photoshop. This digitized image is available for qualitative analysis. An unprocessed, high resolution version of this image (CIL:13123) is in the library and available for quantitative analysis. Additional information available at (http://www5.pbrc.hawaii.edu/allen/).
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