Monoclonal antibody for antigen B2 (called mAb PC-1 in Fok et al., Eur. J. Cell Biol. 40:1-8, 1986) is located on the cytopharynx and along the cytopharyngeal ribbons. It can be seen here on the DV-I membrane that is detached from the cytopharynx. Some acidosomes have already fused with this phagosome. The unfused acidosomes are not labeled with this mAb. TEM taken on 4/3/87 by R. Allen with Zeiss 10A operating at 80kV. Neg. 12,000X. Part published in J. Cell Sci. 108:1263-1274, 1995. Adapted with permission.
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 raw negative was scanned with an Epson Perfection V750 Pro and this high resolution image is best used for quantitative analysis. Additional information available at (http://www5.pbrc.hawaii.edu/allen/).
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