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Scanning electron microscopic study of immunogold-labeled human leukocytes
E H Helinski1, G H Bootsma, R J McGroarty
1Department of Molecular Immunology, Roswell Park Memorial Institute, Buffalo, New York 14263.
Journal of Electron Microscopy Technique
|April 1, 1990
Summary
A new, safe, and cost-effective method using Peldri II for cell preparation in scanning electron microscopy (SEM) is presented. This Peldri II sublimation technique offers advantages over traditional critical point drying (CPD) for visualizing cell surface details.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Materials Science
Background:
- Critical point drying (CPD) is the standard method for preparing cells for scanning electron microscopy (SEM).
- CPD involves complex procedures and specialized equipment.
- There is a need for simpler, safer, and more cost-effective alternatives for SEM specimen preparation.
Purpose of the Study:
- To introduce and evaluate Peldri II, a novel sublimation dehydrant, as an alternative to CPD for SEM.
- To demonstrate the efficacy of Peldri II in preserving cellular morphology and surface features.
- To showcase the application of Peldri II in immunogold labeling studies for visualizing cell surface receptors.
Main Methods:
- Peldri II, a fluorocarbon compound, was used as a sublimation dehydrant for processing various cell types, including leukocytes and lymphoblastoid cell lines.
- Specimens were prepared using the Peldri II procedure and examined using scanning electron microscopy (SEM).
- Immunogold labeling with antibodies targeting the interleukin-2 receptor (IL-2R) was performed to visualize receptor distribution on leukocyte surfaces.
Main Results:
- The Peldri II procedure proved to be simple, efficient, inexpensive, reproducible, and safe.
- SEM imaging revealed well-preserved cellular structures and surface details comparable to CPD.
- Successful visualization of interleukin-2 receptor (IL-2R) expression and distribution on leukocyte membranes using immunogold labeling was achieved.
Conclusions:
- Peldri II offers a viable and advantageous alternative to critical point drying (CPD) for SEM specimen preparation.
- The Peldri II method simplifies the SEM workflow and maintains high-quality imaging of cellular ultrastructure.
- This technique is particularly useful for immunolabeling studies requiring detailed surface analysis of cells.