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Updated: May 13, 2026
09:46
A Method for Obtaining Serial Ultrathin Sections of Microorganisms in Transmission Electron Microscopy
Published on: January 17, 2018
A novel sample preparation method that enables nucleic acid analysis from ultrathin sections
Vincent P Klink1, Giselle Thibaudeau, Ronald Altig
1Department of Biological Sciences, Mississippi State University, Mississippi State, MS 39762, USA. vklink@biology.msstate.edu
Summary
This study introduces a new method for preparing ultrathin cell sections for nucleic acid analysis using laser capture microdissection. The technique successfully preserves cell integrity, enabling RNA isolation from small tissue samples.
Area of Science:
- Developmental Biology
- Cell Biology
- Molecular Biology
Background:
- Nucleic acid analysis of individual cells is crucial for understanding cell development.
- Existing methods may require larger sample sizes, limiting cell-specific studies.
Purpose of the Study:
- To present a novel biological sample preparation method for laser capture microdissection-assisted nucleic acid analysis.
- To evaluate the suitability of Technovit® 9100® as an embedding medium for ultrathin cell/tissue sections.
Main Methods:
- Cells from the mitotic bed of Lithobates sphenocephalus tadpole teeth were isolated and embedded in Technovit® 9100®.
- Thin sections were examined using bright-field and transmission electron microscopy.
- Laser capture microdissection (LCM) was used to isolate developmental series of tooth primordia for RNA analysis.
Main Results:
- The Technovit® 9100® embedding method preserved cytological and ultrastructural integrity of cells.
- Intact RNA was successfully isolated from cells prepared using this method.
- The technique requires minimal sample material, ideal for limited cell quantities.
Conclusions:
- Technovit® 9100® is an effective embedding medium for preparing small tissues and individual cells for LCM and nucleic acid analysis.
- This method facilitates cell-specific molecular studies, particularly when sample material is scarce.
- The developed technique advances the ability to perform detailed analyses on individual cells during development.
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