ArticlebioRxiv : the preprint server for biology2025
Fix or Freeze? Spectral Differences Arising from Tissue Preparation in Chemical Imaging.
Article in bioRxiv : the preprint server for biology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Spectrochemical imaging has emerged as a powerful, label-free modality for visualizing the biochemical composition of tissues based on intrinsic vibrational signatures. Specifically, mid-infrared spectrochemical imaging (MIRSI) is becoming essential for fundamental biomedical research studying disease mechanisms, identifying biomarkers, and guiding drug development. However, the sensitivity of MIRSI to sample preparation protocols and its impact on spectral data interpretation remain poorly characterized. Here, we systematically compared spectral data collected from rat kidney and liver tissues prepared using standard fresh frozen (FF) and formalin-fixed, paraffin-embedded (FFPE) tissue processing methods using quantum cascade laser (QCL)-based MIRSI. We applied frequently used spectral data processing techniques, including uniform manifold approximation and projection (UMAP), correlation matrices, and second-derivative spectral analysis, to characterize preparation-induced differences. FF samples preserved a broader range of biochemical signals, retaining the innate chemical composition of tissues, while FFPE tissues showed reduced spectral diversity and absorption signal intensity. Moreover, a unique spectral band at 1026 cm
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