Hostname: page-component-cd9895bd7-gxg78 Total loading time: 0 Render date: 2024-12-27T09:26:59.629Z Has data issue: false hasContentIssue false

The Use of Poly-L-lysine as an Adhesive in Scanning Electron Microscopy

Published online by Cambridge University Press:  14 March 2018

Jeannette Taylor*
Affiliation:
Emory University, Atlanta, GA

Extract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Poly-L-lysine of a medium molecular weight, 30,000 to 70,000 Daltons (Sigma–Aldrich), is currently used in our facility as a polycationic adhesive on 5 mm square silicon chips to secure individual anionic cells and particles for ease of processing and viewing in the scanning electron microscope. In this technique, a solution of 1 mg/ml of poly-L-lysine is dissolved in distilled water. Drops of the solution are placed onto clean 5 mm square silicon chips and allowed to sit for one to several hours before being wicked away. The prepared chips are used immediately. Suspended cells are then applied to the chips and allowed to settle and adhere, after which they are washed and fixed. Alternatively, fixed suspended cells or particles are applied to these silicon chips. The cells and particles secured to the silicon chips are easily handled through processing steps such as dehydration, critical point drying, and metal sputter coating for viewing in the scanning electron microscope. Presented herein is a brief history of the evolution of the technique of using poly-L-lysine as an adhesive for SEM.

Type
Research Article
Copyright
Copyright © Microscopy Society of America 2008

References

Steinhardt, R.A., Lundin, L., and Mazia, D., “Bioelectric Responses of the Echinoderm Egg to Fertilization”, Proc. Nat. Acad. Sci., Vol. 68, No. 10, Oct. 1971, pp. 24262430.Google Scholar
Alvaro Macieira-Coelho and Stratis Avrameas, “Modulation of Cell Behavior In Vitro by the Substratum in Fibroblastic and Leukemic Mouse Lines”, Proc. Nat. Acad. Sci., Vol. 9, Sept. 1972, pp. 2469-2473.Google Scholar
Mazia, Daniel, Schatten, Gerald, and Sale, Winfield, “Adhesion of Cells to Surfaces Coated with Poly-L-Lysine, Applications to Electron Microscopy”, Journal of Cell Biology, Vol. 66, 1975, pp. 198-200.Google Scholar
Clarke, M., Schatten, G., Mazia, D., and Spudich, J. A., “Visualization of Actin Fibers Associated with the Cell Membrane in Amoebae of Dictyostelium discoideum”, Proc. Nat. Acad. Sci., Vol. 72, No. 5, May 1975, pp. 1758-1762.CrossRefGoogle Scholar
Sanders, S. K., Alexander, E. L., and Braylan, R. C., “A High-Yield Technique for Preparing Cells Fixed in Suspension for Scanning Electron Microscopy”, The Journal of Cell Biology, Vol. 67, 1975, pp. 476-480.Google Scholar
Male, P. M., and Biemesderfer, D., “Silicon Wafers as Support for Biological Macromolecules in High Resolution Cold Field Emission Scanning Electron Microscopy”, Scanning Electron Microscopy, Vol. II, 1978, pp. 643-648. Sigma-Aldrich, Product Information on Poly-L-lysine Hydrobromide, Saint Louis, Missouri, USA, catalogue on the webGoogle Scholar