Astaxanthin as a new Raman probe for biosensing of specific subcellular lipidic structures: can we detect lipids in cell
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Cellular and Molecular Life Sciences
ORIGINAL ARTICLE
Astaxanthin as a new Raman probe for biosensing of specific subcellular lipidic structures: can we detect lipids in cells under resonance conditions? Krzysztof Czamara1 · Adriana Adamczyk1 · Marta Stojak1 · Basseem Radwan1 · Malgorzata Baranska1,2 Received: 2 July 2020 / Revised: 26 October 2020 / Accepted: 19 November 2020 © The Author(s) 2020
Abstract Here we report a new Raman probe for cellular studies on lipids detection and distribution. It is (3S, 3’S)-astaxanthin (AXT), a natural xanthophyll of hydrophobic properties and high solubility in lipids. It contains a chromophore group, a long polyene chain of eleven conjugated C=C bonds including two in the terminal rings, absorbing light in the visible range that coincides with the excitation of lasers commonly used in Raman spectroscopy for studying of biological samples. Depending on the laser, resonance (excitation in the visible range) or pre-resonance (the near infrared range) Raman spectrum of astaxanthin is dominated by bands at ca. 1008, 1158, and 1520 cm−1 that now can be also a marker of lipids distribution in the cells. We showed that AXT accumulates in lipidic structures of endothelial cells in time-dependent manner that provides possibility to visualize e.g. endoplasmic reticulum, as well as nuclear envelope. As a non-toxic reporter, it has a potential in the future studies on e.g. nucleus membranes damage in live cells in a very short measuring time. Keyword Carotenoids · Endothelium · Raman spectroscopy · TNF-α · Inflammation · Lipids · Raman imaging · Fluorescence imaging
Introduction Visualization of subcellular compartments gives valuable insight into cellular processes, which is important for detection and tracking the changes that occur during various cellular events such as cell signalling, metabolism as well as pathology development, consequently, enhancing diagnosis and treatment [1, 2]. Studies on subcellular lipidic structures such as lipid droplets (LDs), endoplasmic reticulum (ER) and nuclear envelope have gained substantial importance due to their roles in many aspects related to health and diseases development [3, 4]. Lipids are crucial for maintaining proper cell functioning by participating in cellular signalling, building membranes, * Malgorzata Baranska [email protected] 1
Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, 14 Bobrzynskiego Str., 30‑ 348 Krakow, Poland
Faculty of Chemistry, Jagiellonian University, 2 Gronostajowa Str., 30‑387 Krakow, Poland
2
and being an energy reservoir. Abnormalities of lipids content, distribution and structure can be a marker of pathological processes undergoing in the cells, e.g. inflammation or development of various lifestyle diseases [5–7]. The idea to use a different type of conjugated probes, e.g. fluorophores (rhodamine labelled phosphatidylethanolamine), hydrophobic dyes like BODIPY (boron dipyrromethene), Nile red or Oil red O (ORO), is to visualise lipids distribution in cells or
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