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Preparing Adherent Cells for X-ray Fluorescence Imaging by Chemical Fixation
Published on: March 12, 2015
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Rapid preparation of adherent mammalian cells for basic scanning electron microscopy (SEM) analysis
Andrew Osahor1, Karthik Deekonda1, Choon-Weng Lee2
1School of Science, Monash University Malaysia, Bandar Sunway 47500, Selangor Darul Ehsan, Malaysia.
Analytical Biochemistry
|July 12, 2017
Summary
This study presents a simplified, cost-effective method for scanning electron microscopy sample preparation, eliminating hazardous reagents and complex equipment. The new protocol enables clear visualization of cellular structures for routine morphological studies.
Area of Science:
- Microscopy
- Cell Biology
- Microbiology
Background:
- Scanning electron microscopy (SEM) sample preparation traditionally uses expensive and hazardous reagents like osmium tetroxide.
- Critical point drying is a complex and time-consuming step in conventional SEM preparation.
Purpose of the Study:
- To develop a simplified, cost-effective, and faster SEM sample preparation protocol.
- To demonstrate the efficacy of the new protocol for visualizing cellular interactions and structures.
Main Methods:
- The study circumvented the use of osmium tetroxide and critical point drying.
- A novel protocol was developed for preparing biological samples for SEM analysis.
Main Results:
- The new protocol significantly reduced the duration, complexity, and cost of SEM sample preparation.
- Early-stage interactions between invasive bacteria and HeLa cells were clearly visualized.
- Sufficient clarity was achieved to preserve and visualize relevant cellular structures.
Conclusions:
- This simplified SEM sample preparation method is cheaper and easier to adapt than conventional techniques.
- The protocol allows for routine preparation and viewing of eukaryotic and bacterial samples for basic morphological studies.
