TY - JOUR

T1 - 4π light scattering flow cytometry: enhancing the identification and characterization of individual cells

AU - Alexandrov, Evgeniy A

AU - Litvinenko, Alena L

AU - Yastrebova, Ekaterina S

AU - Strokotov, Dmitry I

AU - Nekrasov, Vyacheslav M

AU - Gilev, Konstantin V

AU - Chernyshev, Andrei V

AU - Karpenko, Andrey A

AU - Maltsev, Valeri P

N1 - The work was supported by Russian Science Foundation, theoretical simulation and data processing via Grant No. 23-25-00347.

PY - 2023/10/12

Y1 - 2023/10/12

N2 - The analysis of individual particles with complex morphologies from light scattering is crucial in disperse systems studies, such as blood cells. Characterization, which assumes determining particle characteristics, has a higher likelihood of succeeding in solving the inverse light-scattering problem if an instrument provides enough light-scattering data. In this study, we demonstrate how we extend the operating angular interval for the 4π Scanning Flow Cytometer (4πSFC), which measures angle-resolved light-scattering profiles (LSPs) of individual particles. The angular interval is extended by additionally measuring light scattering for the backward hemisphere. Currently, the 4πSFC setup uses three lasers, a single optical cell, and three photomultipliers. It enables the measurement of the LSP of individual particles within the angular interval of 10 to 170° for polar angles with integration over azimuth angles, which covers the spatial angle of 98.5% of the 4π angle. We demonstrate the 4πSFC's performance in measuring LSPs from the analysis of polymer beads, mature and spherized erythrocytes, and platelets. The 4πSFC has the potential to be very useful in identifying platelet dimers and granulocytes without labels, characterizing lymphocytes, monocytes, and abnormal erythrocytes.

AB - The analysis of individual particles with complex morphologies from light scattering is crucial in disperse systems studies, such as blood cells. Characterization, which assumes determining particle characteristics, has a higher likelihood of succeeding in solving the inverse light-scattering problem if an instrument provides enough light-scattering data. In this study, we demonstrate how we extend the operating angular interval for the 4π Scanning Flow Cytometer (4πSFC), which measures angle-resolved light-scattering profiles (LSPs) of individual particles. The angular interval is extended by additionally measuring light scattering for the backward hemisphere. Currently, the 4πSFC setup uses three lasers, a single optical cell, and three photomultipliers. It enables the measurement of the LSP of individual particles within the angular interval of 10 to 170° for polar angles with integration over azimuth angles, which covers the spatial angle of 98.5% of the 4π angle. We demonstrate the 4πSFC's performance in measuring LSPs from the analysis of polymer beads, mature and spherized erythrocytes, and platelets. The 4πSFC has the potential to be very useful in identifying platelet dimers and granulocytes without labels, characterizing lymphocytes, monocytes, and abnormal erythrocytes.

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85173973573&origin=inward&txGid=001b6dd2213d657754fd5b59d7d85f0d

U2 - 10.1039/d3ay01171b

DO - 10.1039/d3ay01171b

M3 - Article

C2 - 37781887

VL - 15

SP - 5218

EP - 5224

JO - Analytical Methods

JF - Analytical Methods

SN - 1759-9660

IS - 39

ER -