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Morphological changes in isolated lymphocytes during preparation for SEM: freeze drying versus critical-point drying
The American Journal of Anatomy
|July 1, 1978
Summary
Freeze drying (FD) preserves lymphocyte size and surface structure better than critical-point drying (CPD) for scanning electron microscopy (SEM). FD lymphocytes are larger with more complex microvilli, crucial for accurate SEM studies.
Area of Science:
- Cell Biology
- Microscopy Techniques
Background:
- Scanning electron microscopy (SEM) requires specific sample preparation techniques.
- Drying methods critically influence the preservation of cellular morphology.
Purpose of the Study:
- To quantitatively compare the effects of critical-point drying (CPD) and freeze drying (FD) on isolated lymphocyte morphology for SEM.
- To determine which drying method best preserves lymphocyte size and surface features.
Main Methods:
- Isolated lymphocytes were prepared using both CPD and FD methods after fixation.
- Cellular diameter and microvilli characteristics (width, number) were quantitatively measured.
- Lymphocyte surfaces were analyzed using SEM.
Main Results:
- FD preserved lymphocyte size better (6.9 micron) compared to CPD (5.7 micron).
- FD resulted in larger, more complex microvilli (0.22 micron width) than CPD (0.12 micron width).
- Microvillus density per cell was similar between both methods.
Conclusions:
- Freeze drying (FD) is superior to critical-point drying (CPD) for preserving lymphocyte size and surface ultrastructure in SEM.
- Method selection is critical for accurate SEM analysis of cellular morphology.
- Researchers should validate drying methods for specific cell types.