Abstract
Minimizing morbidities and mortalities associated with skin cancers requires sustained research with the goal of obtaining fresh insights into disease onset and progression under specific stimuli, particularly the influence of ultraviolet rays. In the present study, label-free profiling of skin fibroblasts exposed to time-bound ultra-violet radiation has been performed using quantitative phase imaging and Raman spectroscopy. Statistically significant differences in quantifiable biophysical parameters, such as matter density and cell dry mass, were observed with phase imaging. Accurate estimation of changes in the biochemical constituents, notably nucleic acids and proteins, was demonstrated through a combination of Raman spectroscopy and multivariate analysis of spectral patterns. Overall, the findings of this study demonstrate the promise of these non-perturbative optical modalities in accurately identifying cellular phenotypes and responses to external stimuli by combining molecular and biophysical information.
Highlights
Quantitative, label-free profiling of cellular phenotypes, during malignant transformation, remains an outstanding challenge in biomedical imaging
It is generally hypothesized that initiation of skin pathologies due to prolonged exposure to sun is caused by formation of free radicals, reactive oxygen and nitrogen species (ROS) and mobilization of transition metal ions leading to impairment of DNA damage repair pathways[27, 29]
Our present study aims at identifying changes in the structural and biochemical composition of skin cells in response to minimal and time-bound ultraviolet radiation (UVR) exposure in a label-free manner
Summary
Quantitative, label-free profiling of cellular phenotypes, during malignant transformation, remains an outstanding challenge in biomedical imaging. Novel approaches relying on global analysis of cellular features, from morphology to the composite biomolecular status (notably chemical composition and molecular conformation), is a pre-requisite for accurate monitoring of cellular processes In this milieu, Raman spectroscopy (RS) and Quantitative Phase Imaging (QPI) have emerged as powerful non-destructive tools for studying cellular pathophysiology, albeit through markedly different light scattering principles[1,2,3,4,5]. We have used QPI and RS as complementary, live cell analysis tools to evaluate the efficacy in identifying changes induced by minimal time bound UVR exposure in skin fibroblasts Combination of these two techniques, one suited for detection of subtle morphological/biophysical alterations while the other appropriate for capturing molecular perturbations, could pave the way to address issues of label-free monitoring of cellular responses in response to an external stimulus
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