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Optical Redox Imaging of Fixed Unstained Muscle Slides Reveals Useful Biological Information
He N Xu1,2,3,4, Huaqing Zhao5, Karthikeyani Chellappa6
1Department of Radiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. hexu2@pennmedicine.upenn.edu.
Molecular Imaging and Biology
|April 13, 2019
Summary
Optical redox imaging (ORI) of unstained FFPE muscle slides reveals age-related changes in cellular redox status. This technique offers quantitative biological insights for potential biomedical applications.
Area of Science:
- Biomedical Imaging
- Cellular Metabolism
- Histopathology
Background:
- Optical redox imaging (ORI) analyzes cellular autofluorescence from nicotinamide adenine dinucleotide (NADH) and oxidized flavoproteins (Fp).
- ORI is valuable for studying cellular energetics and metabolism, with potential in disease diagnosis.
- Quantitative ORI of formalin-fixed paraffin-embedded (FFPE) unstained tissues for disease biomarkers is underexplored.
Purpose of the Study:
- To investigate the utility of ORI for quantitative analysis of FFPE unstained skeletal muscle slides.
- To determine if ORI can provide relevant biological information, particularly regarding age-related redox differences.
Main Methods:
- ORI was performed on living mouse muscle fibers, frozen slides, and FFPE slides.
- FFPE muscle slides from young, mid-age, and Nampt transgenic mice were compared.
- Paraformaldehyde fixation effects on redox ratio were assessed.
Main Results:
- All sample types (living, frozen, FFPE) showed strong NADH and Fp autofluorescence signals.
- Paraformaldehyde fixation significantly increased the Fp/(NADH + Fp) redox ratio in muscle fibers.
- Mid-age muscles exhibited stronger NADH and Fp signals than young muscles; Nampt mice showed altered NADH and Fp levels compared to controls, with soleus muscles appearing more oxidative than gastrocnemius.
Conclusions:
- This study reports for the first time the quantification of aging effects on redox status using ORI of FFPE unstained muscle slides.
- Quantitative ORI of FFPE unstained slides holds promise for biomedical applications.
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