Abstract
Blood cell analysis is one of the standard clinical tests. Despite the widespread use of exogenous markers for blood cell quantification, label-free optical methods are still of high demand due to their possibility for in vivo application and signal specific to the biochemical state of the cell provided by native fluorophores. Here we report the results of blood cell characterization using label-free fluorescence imaging techniques and flow-cytometry. Autofluorescence parameters of different cell types - white blood cells, red blood cells, erythrophagocytic cells - are assessed and analyzed in terms of molecular heterogeneity and possibilities of differentiation between different cell types in vitro and in vivo.
Highlights
Quantification of white blood cells (WBC) content and subtypes in blood is a routine clinical test
When measuring fluorescence signal from the blood vessels, WBC signal is superimposed on the background signal of blood plasma, and exciting radiation and fluorescence emission are effectively absorbed by RBC
Heterogeneity of fluorescence parameters was assessed using flow cytometry, which allowed for label-free characterization of six cell types
Summary
Quantification of white blood cells (WBC) content and subtypes in blood is a routine clinical test. It allows the fractionation and distributional analysis of immune cells within main subpopulations: granulocytes (polymorphonuclear leukocytes), divided into subsets of neutrophils, eosinophils, basophils, and agranulocytes (mononuclear leukocytes), divided into lymphocytes and monocytes [1]. Alterations of the populational distributions of leukocytes may be observed during such severe disorders as ischemic necrosis (e.g. myocardial infarction), leukemia and several hereditary diseases, which are accompanied by increased number of neutrophils, decrease in lymphocytes and eosinophils, and the decrease in neutrophils count is associated with anemia and several cancer types [1]. Routine analysis of WBC subpopulations is commonly performed using flow cytometry, which allows for fast cell counting and differentiation using light scattering and fluorescence emission, mostly of exogenous stains
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