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Updated: Aug 18, 2025
Rigid Embedding of Fixed and Stained, Whole, Millimeter-Scale Specimens for Section-free 3D Histology by Micro-Computed Tomography
Published on: October 17, 2018
A 1-min double embedding method for small tissue specimens preserves comedone histology and eliminates the need for
De-Tian Xu1,2, Yan Zheng3, Yu Shi1
1Shanghai Skin Disease Hospital, Tongji University Medical School, Shanghai, China.
Background:
It is difficult to preserve the structure and microbial distribution inside comedonal plugs during routine processing.
Objective:
The objective of this study is to determine the optimal method to preserve the comedonal corneum plug structure and inherent microorganisms thereby eliminating the need to perform punch biopsies in relevant studies.
Methods:
Corneum plugs were extracted from comedones of acne vulgaris patients. Primary embedding using either a 2% agarose, 2% agar, 25% gelatin, or 2% agar + 2.5% gelatin solution was subsequently performed and the results compared. The specimens were then fixed, waxed, sectioned, and examined by light, fluorescence, and scanning electron microscopies to observe the structures and microorganisms within the plugs.
Results:
Both the 25% gelatin and 2% agarose solutions successfully preserved the structural integrity of corneum plugs and the inherent microorganisms. When considering other factors such as thermostability, reusability, and convenience, the 25% gelatin solution was the superior choice among the four materials.
Conclusion:
We report a simple and effective method for double embedding comedonal plugs and other small tissue specimens. The technique preserves the structure and microbial distribution in situ within comedonal corneum plugs, eliminates the need for punch biopsies. This method may also be applied to other tiny and fragile tissue specimens, thereby enabling a potentially wide array of future large-scale investigations and alleviated patients' pain.
Insights
Researchers developed a new double embedding method using 25% gelatin to preserve comedonal plugs and their microbes. This technique avoids painful punch biopsies for acne vulgaris studies.
Area of Science:
- Dermatology
- Microbiology
- Histology
Background:
- Preserving the structure and microbial distribution within comedonal plugs is challenging with standard processing.
- Routine methods often compromise the integrity of these delicate specimens.
Purpose of the Study:
- To identify an optimal preservation method for comedonal corneum plugs.
- To maintain the structure and microorganisms within plugs, eliminating the need for punch biopsies.
Main Methods:
- Comedonal plugs were extracted from acne vulgaris patients.
- Primary embedding was performed using agarose, agar, gelatin, or a combination.
- Specimens were processed and examined using light, fluorescence, and scanning electron microscopy.
Main Results:
- Both 25% gelatin and 2% agarose effectively preserved plug structure and microorganisms.
- 25% gelatin demonstrated superior thermostability, reusability, and convenience.
Conclusions:
- A simple, effective double embedding method using 25% gelatin was established.
- This technique preserves in situ structure and microbial distribution in comedonal plugs.
- The method eliminates the need for punch biopsies and can be applied to other small tissue specimens.
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