Abstract
The alteration of the phospholipid composition within the cell, in particular the ratio between saturated and unsaturated fatty acids, can serve as an important biomarker to prognosis of the disease progression (e.g., fatty-liver disease, prostate cancer, or neurodegenerative disorders). Major techniques for lipid analysis in biological samples require a lipid extraction procedure that is not compatible with live cell studies. To address this challenge, we apply microRaman-Biomolecular Component Analysis (BCA) for comparative analysis of phospholipid composition and sensing the saturation degree of fatty acid lipid chain in live HeLa cells and lipids extracted from HeLa cells. After processing raw Raman data, acquired in lipid droplets (LDs) free cytoplasmic area, LDs and extracted lipids with BCA, the lipid component was isolated. Despite the similarity in general profiles of processed Raman spectra acquired in live cells and extracted lipids, some clear differences that reflect diversity in their phospholipids composition were revealed. Furthermore, using the direct relation between the number of double bonds in the fatty acid chain and the intensity ratio of the corresponding Raman bands, the saturation degree of fatty acids was estimated.
Highlights
It is known that lipids play vital roles in establishing cellular architecture and maintaining cellular processes, including but not limited to energy storage, signaling reactions, protection against some forms of cellular stress, and many others [1,2,3]
Aimed to figure the difference in phospholipid composition between different sites in live cells as well as to explore the impact of extraction of lipids procedure on phospholipids content compared with live cells, the phospholipid composition of lipid droplets (LDs) in live cells and lipids extracted from HeLa cells are similar, while
A comparative analysis of processed Raman spectra of LDs, LDs free cytoplasmic area, and extracted lipids revealed that phospholipid content is significantly different in live cells and extracted
Summary
It is known that lipids play vital roles in establishing cellular architecture and maintaining cellular processes, including but not limited to energy storage, signaling reactions, protection against some forms of cellular stress, and many others [1,2,3]. Their dysfunction has been linked to many diseases, e.g., obesity, diabetes, fatty liver disease, autoimmunity, prostate cancer, and some of the neurodegenerative disorders [4,5,6,7,8,9,10].
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