Detection and identification of alginate in tissue-freezing media samples using Raman spectroscopy

Monireh Pourrahimi; Samaneh Ghazanfarpour; Alireza Sheikhsofla; Rafael Pena; Jennifer Morrissey; Sujith Chander Reddy Kollampally; Anna Sharikova; Yubing Xie; Melinda Larsen; Alexander Khmaladze

doi:10.1117/12.3003876

12 March 2024 Detection and identification of alginate in tissue-freezing media samples using Raman spectroscopy

Monireh Pourrahimi, Samaneh Ghazanfarpour, Alireza Sheikhsofla, Rafael Pena, Jennifer Morrissey, Sujith Chander Reddy Kollampally, Anna Sharikova, Yubing Xie, Melinda Larsen, Alexander Khmaladze

Author Affiliations +

Proceedings Volume 12854, Label-free Biomedical Imaging and Sensing (LBIS) 2024; 128540N (2024) https://doi.org/10.1117/12.3003876
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Alginate is a natural polysaccharide found in brown algae and has a unique feature, the ability to form a hydrogel upon encountering Ca²⁺. Its exceptional characteristics make alginate hydrogels highly desirable for a range of biomedical applications, such as drug delivery, wound healing, and in particular, tissue engineering and cell therapy, where it is used as scaffolding or as a cell delivery vehicle. After using alginate hydrogel for cell delivery in vivo, one of our objectives was to specifically detect alginate in mouse tissue cryosections containing cell-scaffold constructs to evaluate scaffold cell-scaffold integration with host tissue and degradation. Due to difficulties encountered in detecting alginate using immunohistochemistry with mouse-derived antibodies, we aimed to develop an alternative method to definitively identify alginate within tissue cryosection samples using Raman spectroscopy. The Raman spectra of pure tissue had specific peaks convenient for identification. We identified a region where alginate consistently had stronger signal than either tissue or tissue freezing media. We also detected alginate-specific Raman peaks at 816, 888, 959, 1309, 1433 cm^-1. By collecting the Raman spectra of the samples containing all three substances (alginate, freezing media, and tissue), analyzing them either by characteristic spectral peaks or classical least squares (CLS) method, and mapping the media, alginate, and tissue on the brightfield sample image, we were able to discriminate the alginate from tissue and freezing media. The notable sensitivity and specificity of Raman spectroscopy renders it a promising method for the identification of alginate and alginate-based materials in tissue engineering.

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(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

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Monireh Pourrahimi, Samaneh Ghazanfarpour, Alireza Sheikhsofla, Rafael Pena, Jennifer Morrissey, Sujith Chander Reddy Kollampally, Anna Sharikova, Yubing Xie, Melinda Larsen, and Alexander Khmaladze "Detection and identification of alginate in tissue-freezing media samples using Raman spectroscopy", Proc. SPIE 12854, Label-free Biomedical Imaging and Sensing (LBIS) 2024, 128540N (12 March 2024); https://doi.org/10.1117/12.3003876

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