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Updated: Aug 8, 2026
Thin Sectioning of Slice Preparations for Immunohistochemistry
Published on: April 28, 2007
An improved method for preparing thick sections for immuno/histochemistry and confocal microscopy and its use to
P Monaghan1, P R Watson, H Cook
1Institute for Animal Health, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK. monaghan@bbsrc.ac.uk
Insights
Imaging thick tissue sections using a vibrating microtome and confocal microscopy enables detection of rare events like bacterial invasion and apoptosis. This method preserves tissue structure and minimizes antigenic damage for accurate analysis.
Area of Science:
- Biomedical Imaging
- Cell Biology
- Histology
Background:
- Detecting rare cellular events in solid tissues is challenging with standard imaging techniques.
- Immunocytochemistry and specific staining methods require optimal sample preservation.
Purpose of the Study:
- To present a method for imaging thick tissue sections for rare event detection.
- To evaluate the effectiveness of this method in preserving tissue structure and antigenicity.
Main Methods:
- Preparation of 40-100 micrometer thick paraformaldehyde-fixed solid tissue sections using a vibrating microtome.
- Immunolabeling of thick sections followed by confocal microscopy.
- TUNEL staining for apoptotic cell nuclei detection.
Main Results:
- The vibrating microtome method allows for excellent preservation of sample structure.
- Minimal antigenic damage was observed, facilitating effective immunolabeling.
- Individual invading Salmonella bacteria in bovine intestinal epithelium were detected.
- Rare apoptotic cell nuclei were successfully identified using TUNEL staining in thick sections.
Conclusions:
- Imaging thick tissue sections prepared by vibrating microtomy is a valuable technique for detecting rare events in solid tissues.
- This approach enhances the sensitivity of immunocytochemistry and specific staining methods.
- The method offers a good balance between sample integrity and detection sensitivity for biological research.
Abstract:
Detection of rare events within solid tissues by immunocytochemistry is aided by imaging thick sections. Sections of 40--100 microm thickness of paraformaldehyde-fixed solid tissue can be prepared by use of a vibrating microtome and when immunolabelled these sections can be imaged in a confocal microscope. This approach provides excellent preservation of the structure of the sample and imposes minimal antigenic damage. In studies of the invasion of the bovine intestinal epithelium by Salmonella, this method has allowed detection of individual invading bacteria within large samples. The thick vibrating microtome sections were also used for the detection of rare apoptotic cell nuclei identified by TUNEL staining.
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