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Updated: May 5, 2026
Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections
Published on: November 14, 2016
A protocol for concurrent high-quality immunohistochemical and biochemical analyses in adult mouse central nervous
Tina Notter1, Patrizia Panzanelli, Sandra Pfister
1Institute of Pharmacology and Toxicology, University of Zurich, CH-8057, Zurich, Switzerland; Neuroscience Center Zurich, Federal Institute of Technology and University of Zurich, Zurich, Switzerland.
Insights
This study introduces a novel protocol for simultaneous biochemical and immunohistochemical analysis of brain tissue. The method preserves tissue integrity, enabling combined analyses and reducing the number of animals needed for research.
Area of Science:
- Neuroscience
- Biochemistry
- Immunohistochemistry
Background:
- Biochemical analysis requires fresh homogenates, while immunohistochemistry uses fixed tissue.
- Current fixation methods compromise morphological preservation and protein antigenicity.
- Concurrent analysis from the same tissue sample is challenging.
Purpose of the Study:
- To develop a versatile protocol for simultaneous biochemical and immunohistochemical analysis from the same animal tissue.
- To improve tissue preservation for both biochemical and morphological assessments.
- To reduce the number of animals required for combined analytical studies.
Main Methods:
- A brief transcardiac perfusion with oxygenated, glucose-supplemented artificial cerebrospinal fluid.
- Isolation of brain regions of interest for subsequent homogenization or fixation.
- Concurrent biochemical assays and immunohistochemistry, including pre-embedding immunoelectron microscopy.
Main Results:
- The protocol allows for optimal biochemical and morphological analysis.
- Enhanced sensitivity and preservation of tissue structure were observed.
- A reduction in artifacts typically seen in perfusion-fixed tissue was achieved.
Conclusions:
- The developed protocol enables concurrent biochemical and immunohistochemical analyses from the same tissue.
- This method offers optimal sensitivity and tissue preservation, minimizing artifacts.
- The protocol is versatile and can reduce animal usage in research.
Abstract:
Biochemical analysis of central nervous system proteins and nucleic acids requires fresh-tissue homogenates, whereas immunohistochemistry usually is performed in sections prepared from perfusion-fixed tissue. Post-mortem immersion-fixation is possible, but largely impairs morphological preservation and protein antigenicity. Here, we present a simple, fast and versatile protocol allowing concurrent biochemical and immunohistochemical analysis, including pre-embedding immunoelectron microscopy, using tissue from the same animal. The protocol includes a brief transcardiac perfusion with ice-cold, oxygenated and glucose-supplemented artificial cerebrospinal fluid to maintain brain tissue alive, prior to isolation of regions of interest, followed by homogenisation for biochemistry or immersion-fixation for immunohistochemistry. We provide several examples demonstrating that this protocol allows optimal biochemical and morphological analysis, characterised with optimal sensitivity and preservation of tissue structure, along with a reduction of artefacts typically seen in perfusion-fixed tissue. This protocol should find widespread applications for combining analytical methods in tissue from the same animal, thereby reducing the number of mice required for a given experiment.
