How to use the microscope to observe with the cutting method

If the sample is embedded in styrene resin for microscopic analysis, it's highly convenient to perform scanning electron microscope (SEM) observations through sectioning. This is because styrene is a polymer that can be easily dissolved by solvents such as propylene oxide and isovaleric acid. By changing the solvent multiple times, the styrene can be effectively removed. When preparing transmission electron microscopy (TEM) samples, several embedded capsules are typically made. Once the sample is fully polymerized, it is cut using the described method. The sample is then placed in propylene oxide for about two hours before being transferred to isocyanuric acid for critical point drying. Besides the resin-based cutting method, there are other techniques such as organic solvent-based methods (e.g., alcohol, isovaleric acid), water-soluble embedding agents (like dimethyl hydrazine or glycerin), and physical methods like freezing or freeze-fracturing. These methods are generally easier to implement. For example, with the alcohol-based method, the sample is first fixed and dehydrated using alcohol. Once dehydration is complete, the sample is placed into a capsule filled with anhydrous ether, ensuring no air bubbles form. The capsule is then placed in a low-temperature environment to cure. Afterward, the sample is sectioned. The cut sample is immediately immersed in anhydrous ethyl ether at room temperature, allowed to melt, and then transferred to isoamyl acetate for critical point drying. It should be noted that specimens prepared using the cutting method require high-resolution imaging, such as with a field emission scanning electron microscope. Standard SEMs have lower resolution compared to TEMs. If you're observing cellular structures like organelles, a field emission electron gun is essential for accurate imaging. Due to the strong three-dimensional effect of SEM, the cut sample reveals the spatial arrangement of structures like the Golgi apparatus, large and small vacuoles, mitochondria appearing as spacers, and the nucleus resembling a small mushroom attached to the endoplasmic reticulum "plate." However, some researchers argue that polarized light microscopy does not offer significant new insights into cell structure, which has limited its widespread use.

Kitchen Storage And Organization

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