Micro Raman spectroscopy for NETosis detection

Abstract

Over the past few years, Raman spectroscopy has become a powerful diagnostic tool in the life sciences. The present work is devoted to the application of Raman microspectroscopy for distinction of neutrophils transformed during NETosis and the quantitative determination of the level of their transformation based on the analysis of the neutrophil Raman spectra acquired from the samples of human blood at different levels of transformation. NETosis is a process of the programmed neutrophil cell death involved in the development of many diseases, including those associated with high mortality. Our goal was to search for possible spectral markers in neutrophil Raman spectra, caused precisely by NETotic transformation of neutrophils. The results of the work were (a) obtaining of neutrophil Raman spectra at different levels of cell transformation; (b) creation of spectral archetypes of neutrophils (as a "representative" Raman spectrum for spectra group) with a given level of cell transformation; (c) detection of statistically significant differences in the spectral archetypes of the neutrophil Raman spectra at different levels of transformation. K E Y W O R D S human blood neutrophils, NETosis, microRaman spectroscopy, spectra preprocessing, spectral marker 1 | INTRODUCTION Raman microspectroscopy is widely used in the studies of cell biology, microbiology, and medicine for optical analysis of biological objects at the cellular level. [1-6] Raman microspectroscopy allows exploring intracellular transformations and their features [7-12] and helps to understand the processes occurring in the cell when studying the properties of the selective interaction of reagents in the cell. [13-15] Recently, achievements in Raman microspectroscopy have opened up new prospects for the rapid and sensitive detection of bacteria of various types. [16-21] Neutrophils are the most common human blood leukocytes, which are the most important part of the innate immunity and carry out a fast response to microbial invasion. In 2004, a new type of programmed cell death of neutrophils, called NETosis, [22] was described. Currently,

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