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A simple technique for examining frozen hydrated specimens in the scanning electron microscope
Journal of Microscopy
|August 1, 1975
Summary
A new scanning electron microscope technique allows simple examination of frozen hydrated specimens. This method prevents charging and dehydration artifacts, enabling long-term observation of large samples.
Area of Science:
- Materials Science
- Microscopy
- Biophysics
Background:
- Examining hydrated biological or material samples using scanning electron microscopy (SEM) is challenging due to potential dehydration and charging artifacts.
- Traditional SEM techniques often require extensive sample preparation, which can alter the native state of hydrated specimens.
Purpose of the Study:
- To present a simplified SEM technique for the direct examination of frozen hydrated specimens.
- To demonstrate the capability of the technique in preventing common artifacts like charging and dehydration distortion.
Main Methods:
- Utilized a scanning electron microscope equipped with a wide-angle backscattered electron detector.
- Implemented independent control of specimen chamber pressure separate from column pressure.
- Examined large frozen hydrated specimens with minimal preparation time.
Main Results:
- Successfully examined large specimens within one minute of placement.
- Observed specimens for extended periods without detrimental charging artifacts.
- Prevented specimen distortion typically caused by dehydration during SEM analysis.
Conclusions:
- The described SEM technique offers a straightforward and effective method for analyzing frozen hydrated samples.
- Independent control of specimen chamber pressure is crucial for mitigating artifacts in hydrated sample examination.
- This approach enhances the ability to study the native structure of hydrated specimens over time.