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[Scanning electronic microscopy of the myocardium: alternative technics]
A Zepeda Rodríguez1, M E Miranda Martínez, D Cruz Robles
1Departamento de Patología, Instituto Nacional de Cardiología Ignacio Chávez, INCICH, México, D.F.
Archivos Del Instituto De Cardiologia De Mexico
|September 1, 1994
Summary
This study introduces a new method using osmium-thiocarbohydrazide-osmium (OTO) with Peldri II or hexamethyldisilazane (HMDS) to enhance biological specimen imaging in scanning electron microscopy (SEM). Both new techniques provide high-quality SEM images of rat myocardium.
Area of Science:
- Biological Sciences
- Microscopy
- Materials Science
Background:
- Biological specimens often lack electron conductivity, necessitating conductive coating for scanning electron microscopy (SEM).
- Standard coating methods can sometimes obscure fine details or damage delicate biological structures under electron beam irradiation.
Purpose of the Study:
- To evaluate an alternative SEM preparation technique combining osmium-thiocarbohydrazide-osmium (OTO) with Peldri II or hexamethyldisilazane (HMDS).
- To improve the imaging quality of biological specimens, specifically freeze-fractured rat myocardium, for SEM analysis.
Main Methods:
- Freeze-fractured rat myocardium specimens were prepared using three methods: standard technique, OTO with Peldri II, and OTO with HMDS.
- All specimens underwent fixation with cacodylate buffer and glutaraldehyde, followed by examination using scanning electron microscopy (SEM).
Main Results:
- Standard preparation allowed visualization of myocytes and subcellular components like nucleus, myofibrils, and mitochondria.
- Specimens treated with OTO and HMDS, or OTO and Peldri II, yielded high-quality SEM images.
- The alternative methods effectively improved the visualization of biological ultrastructure.
Conclusions:
- The combined OTO with Peldri II or HMDS offers a viable alternative for preparing biological specimens for SEM.
- These enhanced preparation techniques can lead to high-quality imaging, aiding in the detailed study of cellular and subcellular structures.
- This method may be beneficial for researchers seeking improved SEM imaging of biological tissues.