Immunogold label specific for Q2 antigen on discoidal vesicles. The same mAb used in CIL:36721 heavily labels discoidal vesicles and less heavily the cytopharyngeal membrane. A few gold particles are also found on the acidosomes. The distribution of this label confirms that the discoidal vesicles fuse with the cytopharynx to form the phagosome. TEM taken on 10/24/92 by R. Allen with Zeiss 10A operating at 80kV. Neg. 12,000X. Bar = 0.5µm.
To label membranes inside the cell we used very lightly fixed cells (0.25% glutaraldehyde) that were then rapidly frozen in liquid nitrogen and sectioned later at -100oC. These frozen sections were picked up on drops of methylcellulose and transferred to a Formvar-supported grid. The sections were immunogold labeled (15nm gold) to show the location of the specific antigen inside the cell as well as on the cell surface. Microtome sections prepared at approximately 75nm thickness. The negative was printed to paper and the image was scanned to Photoshop. This image is suitable for qualitative analysis. A high resolution version of this image in the library (CIL:40555) which is available for quantitative analysis. Additional information available at (http://www5.pbrc.hawaii.edu/allen/).
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