A high resolution micrograph of the collecting canal of the contractile vacuole surrounded by smooth spongiome remnants and decorated tubules of the CVC. Microtubules bordering the flattened collecting canal are labeled with immunogold when thin frozen sections of the CVC are exposed by the indirect immunogold technique to show anti-b-tubulin. These microtubules have their origin at the cytosolic side of the CV pore. TEM taken on 2/25/94 by R. Allen with Zeiss 10A operating at 80kV. Neg. 9,780X. 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 an Epson Perfection V750 Pro. This image is best used for quantitative analysis. Additional information available at (http://www5.pbrc.hawaii.edu/allen/).
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