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Updated: Sep 20, 2026
An Alternative Approach for Sample Preparation with Low Cell Number for TEM Analysis
Published on: October 12, 2016
Development of preparing small-amount samples for transmission electron microscopy
Y Kaji1, K Kawamoto, S Wakisaka
1Department of Neurosurgery, Miyazaki Medical College.
Abstract:
The means of examination of small-amount samples by transmission electron microscopy (TEM) is described. In this study, the amount of cells necessary for TEM was examined by using a KMU-100 cultured cell line. Floating cells diluted with phosphate buffer solution (PBS) to several concentrations, were gathered in the tip of Beem capsules by means of centrifuge. After fixation, dehydration, and embedding with Epon 812, the samples were cut with a diamond knife, followed by electron staining, and examined by TEM. As a result, we learned that an amount of cells over 1 x 10(3) could be observed by TEM. In spite of cell damage through this procedure, the microscopic structures of the cells were relatively maintained compared with the original cell monolayers. In the near future, it may be possible to examine the sorted cells by flow cytometry (FCM) with TEM.
Insights
Researchers determined the minimum cell quantity for transmission electron microscopy (TEM). Over 1 x 10^3 cells are sufficient for TEM, preserving cellular structures for detailed analysis.
Area of Science:
- Cell Biology
- Microscopy Techniques
Background:
- Transmission electron microscopy (TEM) requires specific sample preparation.
- Determining the minimum sample size is crucial for efficient analysis.
Purpose of the Study:
- To establish the minimum number of cells required for successful TEM examination.
- To evaluate the preservation of cellular structures in small-amount samples.
Main Methods:
- Cultured cells (KMU-100) were diluted to various concentrations in phosphate-buffered saline (PBS).
- Cells were collected via centrifugation, fixed, dehydrated, and embedded in Epon 812.
- Samples were sectioned, stained, and analyzed using TEM.
Main Results:
- A minimum of 1 x 10^3 cells were observable using TEM.
- Despite procedural damage, cellular microscopic structures were relatively maintained compared to cell monolayers.
Conclusions:
- TEM can effectively analyze small cell samples, requiring over 1 x 10^3 cells.
- This method allows for the examination of cellular ultrastructure from limited cell populations.
Related Concept Videos
01:15Transmission Electron Microscopy
01:20Preparation of Samples for Electron Microscopy