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A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
Published on: March 30, 2020
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Simultaneous In Situ Detection of Ferroptosis-Related Multiple Molecules in Tissue Slices Using Thin-Layer
Tatsuya Nishihara1, Rika Tsukame1, Kazuki Bando2,3
1Department of Chemistry and Biological Science, College of Science and Engineering, Aoyama Gakuin University, 5-10-1 Fuchinobe, Chuo-ku, Sagamihara 252-5258, Japan.
ACS Sensors
|November 10, 2025
Summary
Researchers developed a new Raman spectroscopy method for detecting ferroptosis markers like glutathione (GSH) and iron (Fe2+) in tissues. This technique enables simultaneous in situ analysis, aiding ferroptosis research.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Spectroscopy
Background:
- In situ detection of glutathione (GSH) and Fe2+ in tissues is challenging due to their chemical instability.
- Conventional mass spectrometry imaging faces limitations in analyzing these ferroptosis-related molecules.
Purpose of the Study:
- To develop a method for simultaneous in situ detection of ferroptosis-related molecules in frozen tissue sections.
- To utilize Raman spectroscopy for multicomponent analysis of biological samples.
Main Methods:
- Developed a thin-layer chromatography (TLC) plate functionalized with Raman-tag probes (alkynes, nitriles).
- Attached frozen tissue sections to the functionalized TLC plate for GSH addition reactions and iron complex formation.
- Analyzed Raman spectra of TLC plates post-separation to detect target molecules.
Main Results:
- Successfully detected signals of probes in the presence of GSH and Fe2+ targets.
- Tracked changes in GSH and iron ion levels in mouse liver tissue.
- Observed a significant decrease in GSH and an increasing trend in iron ions during acetaminophen-induced ferroptosis.
Conclusions:
- The developed analytical platform enables powerful in situ study of ferroptosis-related molecule dynamics in tissues.
- This method has broad applications in basic research and clinical diagnostics for ferroptosis.
- The findings open new avenues for multiplexed molecular detection in biological tissues.
