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Updated: Apr 1, 2026
Use of Synaptic Zinc Histochemistry to Reveal Different Regions and Laminae in the Developing and Adult Brain
Published on: October 29, 2017
Immunohistochemical demonstration of specific antigens in the human brain fixed in zinc-ethanol-formaldehyde
D E Korzhevskii1, E G Sukhorukova, O V Kirik
1Laboratory of Functional Morphology of the Central and Peripheral Nervous System, Institute of Experimental Medicine. dek2@yandex.ru.
Insights
Zinc-ethanol-formalin (ZEF) fixation is effective for human brain histology and immunohistochemistry. This method preserves morphology and allows for the detection of 11 key antigens in brain tissue.
Area of Science:
- Neuroscience
- Histology
- Biochemistry
Background:
- Tissue fixation is a crucial step in preparing samples for histological analysis and immunohistochemistry.
- Traditional fixatives like formalin can sometimes compromise antigenicity or tissue morphology.
- A novel zinc-ethanol-formalin (ZEF) fixative was developed to potentially overcome these limitations.
Purpose of the Study:
- To evaluate the efficacy of the zinc-ethanol-formalin (ZEF) fixative for human brain tissue.
- To assess the preservation of human brain morphology using ZEF fixation.
- To determine the detectability of various antigens in human brain specimens fixed with ZEF.
Main Methods:
- Human brain tissue samples were fixed using the zinc-ethanol-formalin (ZEF) method.
- Standard histological techniques were applied to assess tissue morphology.
- Immunohistochemistry was performed to evaluate the preservation and detectability of 11 specific antigens.
Main Results:
- The ZEF fixative demonstrated suitability for general histological practice, enabling detailed characterization of human brain morphology.
- All 11 tested antigens, including neuronal markers (NeuN, synaptophysin) and glial markers (GFAP, Iba-1), were well-preserved in ZEF-fixed human brain specimens.
- ZEF fixation facilitated reliable antigen detection, indicating good antigenicity retention.
Conclusions:
- Zinc-ethanol-formalin (ZEF) fixation is a viable method for human brain histology, offering excellent morphological detail.
- ZEF fixation effectively preserves antigenicity, allowing for robust immunohistochemical analysis of various neuronal and glial markers in the human brain.
- This fixative shows significant potential for routine use in neuropathology and neuroscience research.
Abstract:
Tissue fixation is critical for immunohistochemistry. Recently, we developed a zinc-ethanol-formalin fixative (ZEF), and the present study was aimed to assess the applicability of the ZEF for the human brain histology and immunohistochemistry and to evaluate the detectability of different antigens in the human brain fixed with ZEF. In total, 11 antigens were tested, including NeuN, neuron-specific enolase, GFAP, Iba-1, calbindin, calretinin, choline acetyltransferase, glutamic acid decarboxylase (GAD65), tyrosine hydroxylase, synaptophysin, and α-tubulin. The obtained data show that: i) the ZEF has potential for use in general histological practice, where detailed characterization of human brain morphology is needed; ii) the antigens tested are well-preserved in the human brain specimens fixed in the ZEF.
