G-protein coupled short-chain fatty acid receptor GPR 43 fluorescence immunohistochemistry staining protocol for confocal microscopy in human colon cancer

Ilyés Tamás; Éva Korpos; Monica Focșan; Andrei Flaviu Zaharie; Simona Roxana Gheorghe; Ciprian Nicolae Silaghi; Alexandra Mărioara Crăciun

doi:10.2478/rrlm-2026-0012

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G-protein coupled short-chain fatty acid receptor GPR 43 fluorescence immunohistochemistry staining protocol for confocal microscopy in human colon cancer

Revista Romana de Medicina de Laborator

Volume 34 (2026): Issue 2 (April 2026)

By: Ilyés Tamás, Éva Korpos, Monica Focșan, Andrei Flaviu Zaharie, Simona Roxana Gheorghe, Ciprian Nicolae Silaghi and Alexandra Mărioara Crăciun

Open Access

|Apr 2026

Abstract

Background

G-protein coupled receptor 43 (GPR43) mediates the biological effects of short-chain fatty acids (SCFAs) in the colon, but its role in colon cancer (CC) remains insufficiently characterized. This pilot study presents in detail a cost-effective and easily reproducible fluorescence immunohistochemistry protocol for staining GPR43 in formaldehyde-fixed, paraffin-embedded human colon tissue.

Methods

Using samples from patients with colon adenocarcinoma (ACC) and signet ring cell carcinoma (SRCC), we optimized staining procedures with generic reagents and a custom 3D-printed slide tray, facilitating minimal reagent usage and consistent sample coverage.

Results

Confocal microscopy revealed distinct GPR43 localization on the apical and basal membranes of colonocytes and goblet cells in healthy tissue. In contrast, ACC tissue exhibited diffuse, disorganized staining, while SRCC samples showed minimal receptor expression. These findings suggest a link between GPR43 expression patterns and tissue differentiation status. Additionally, we observed autofluorescence at 488 nm of eosinophils without staining, as previously reported in literature.

Conclusions

These findings may support prognostic assessments based on intratumoral or peritumoral eosinophil counts. The present study shows in detail an easily reproducible IHC staining method for GPR 43 in paraffin embedded tissue samples, using only generic materials and highlights its potential role as a biomarker or therapeutic target in CC.

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Articles in this issue

DOI: https://doi.org/10.2478/rrlm-2026-0012 | Journal eISSN: 2284-5623 | Journal ISSN: 1841-6624

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Language: English

Page range: 157 - 166

Submitted on: Nov 22, 2025

Accepted on: Feb 10, 2026

Published on: Apr 24, 2026

Published by: Romanian Association of Laboratory Medicine

In partnership with: Paradigm Publishing Services

Publication frequency: 4 issues per year

Keywords:

colon cancer,

confocal microscopy,

GPR43,

immunohistochemistry,

short-chain fatty acids

Related subjects:

Microbiology and virology

© 2026 Ilyés Tamás, Éva Korpos, Monica Focșan, Andrei Flaviu Zaharie, Simona Roxana Gheorghe, Ciprian Nicolae Silaghi, Alexandra Mărioara Crăciun, published by Romanian Association of Laboratory Medicine
This work is licensed under the Creative Commons Attribution 4.0 License.

Volume 34 (2026): Issue 2 (April 2026)