Description

High resolution view of the contractile vacuole pore encircled on its cytosolic surface with microtubules. Other microtubules arise from these encircling microtubules and extend far out into the cell where they provide structural support for the several radial arms of the CVC. This CV was in very late systole or very early diastole. b-tubulin is immunogold labeled. TEM taken on 7/5/96 by R. Allen with Zeiss 10A operating at 80kV. Neg. 9,780X.

Technical Details

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 detailed in Methods Cell Biol. 2010;96:143-73. The raw film was scanned with a Nikon Coolscan 9000ED. This image is best used for quantitative analysis. Additional information available at (http://www5.pbrc.hawaii.edu/allen/).