Abstract
Human leukocytes fixed in suspension were allowed to settle onto poly-L-lysine-coated glass coverslips and prepared for observation with the scanning electron microscope (SEM). The coverslips were dehydrated in ethanol, critical point dried with CO2, and coated with gold-palladium. With the aid of a locator grid, several fields were photographed with light microscopy after the cells had settled onto the poly-L-lysine-coated coverslips and again after completion of the processing before SEM observation. Quantitative comparison of the number of cells present after settling with the number retained for final viewing with the SEM revealed a cell yield approaching 100%. This simple, reproducible, high-yield technique for processing cells fixed in suspension for SEM prevents changes in surface architecture induced by collecting live cells onto various substrates before fixation and also avoids potentially selective cell losses. Such a technique should allow quantitative correlations between SEM and other morphological and functional parameters.
Full Text
The Full Text of this article is available as a PDF (1.2 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alexander E. L., Wetzel B. Human lymphocytes: similarity of B and T cell surface morphology. Science. 1975 May 16;188(4189):732–734. doi: 10.1126/science.1079095. [DOI] [PubMed] [Google Scholar]
- Linthicum D. S., Sell S. Letter: A rabbit blood lymphocyte is both smooth and hairy. N Engl J Med. 1974 Jun 6;290(23):1327–1328. doi: 10.1056/NEJM197406062902323. [DOI] [PubMed] [Google Scholar]
- Linthicum D. S., Sell S., Wagner R. M., Trefts P. Scanning immunoelectron microscopy of mouse B and T lymphocytes. Nature. 1974 Nov 8;252(5479):173–175. doi: 10.1038/252173a0. [DOI] [PubMed] [Google Scholar]
- Mazia D., Schatten G., Sale W. Adhesion of cells to surfaces coated with polylysine. Applications to electron microscopy. J Cell Biol. 1975 Jul;66(1):198–200. doi: 10.1083/jcb.66.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Polliack A., Lampen N., Clarkson B. D., De Harven E., Bentwich Z., Siegal F. P., Kunkel H. G. Identification of human B and T lymphocytes by scanning electron microscopy. J Exp Med. 1973 Sep 1;138(3):607–624. doi: 10.1084/jem.138.3.607. [DOI] [PMC free article] [PubMed] [Google Scholar]