TY - JOUR

T1 - Ultraviolet light scattering scanning flow cytometry in the characterization of submicron microparticles

AU - Strokotov, Dmitry I

AU - Nekrasov, Vyacheslav M

AU - Gilev, Konstantin V

AU - Karpenko, Andrey A

AU - Maltsev, Valeri P

N1 - Funding information: Russian Science Foundation, Grant/AwardNumber: 17–75-20117. © 2023 International Society for Advancement of Cytometry.

PY - 2023/9

Y1 - 2023/9

N2 - Ultraviolet lasers are commonly used in flow cytometry to excite fluorochrome molecules with subsequent measurement of the specific fluorescence of individual cells. In this study, the performance of the ultraviolet light scattering (UVLS) in the analysis of individual particles with flow cytometry has been demonstrated for the first time. The main advantage of the UVLS relates to the improvement of the analysis of submicron particles due to the strong dependence of the scattering efficiency on the wavelength of the incident light. In this work, submicron particles were analyzed using a scanning flow cytometer (SFC) that allows measurements of light scattering in an angle-resolved regime. The measured light-scattering profiles of individual particles were utilized in solution of the inverse light-scattering problem to retrieve the particle characteristics using a global optimization. The standard polystyrene microspheres were successfully characterized from the analysis of UVLS which provided the size and refractive index (RI) of individual beads. We believe that the main application of UVLS relates to the analysis of microparticles in a serum, in particular in the analysis of chylomicrons (CMs). We have demonstrated the performance of the UVLS SFC in the analysis of CMs of a donor. The RI versus size scatterplot of CMs was successfully retrieved from the analysis. The current set-up of the SFC has allowed us to characterize individual CMs starting from the size of 160 nm that provides determination of the CM concentration in a serum with flow cytometry. This feature of the UVLS should help with the analysis of lipid metabolism measuring RI and size map evolution after lipase action.

AB - Ultraviolet lasers are commonly used in flow cytometry to excite fluorochrome molecules with subsequent measurement of the specific fluorescence of individual cells. In this study, the performance of the ultraviolet light scattering (UVLS) in the analysis of individual particles with flow cytometry has been demonstrated for the first time. The main advantage of the UVLS relates to the improvement of the analysis of submicron particles due to the strong dependence of the scattering efficiency on the wavelength of the incident light. In this work, submicron particles were analyzed using a scanning flow cytometer (SFC) that allows measurements of light scattering in an angle-resolved regime. The measured light-scattering profiles of individual particles were utilized in solution of the inverse light-scattering problem to retrieve the particle characteristics using a global optimization. The standard polystyrene microspheres were successfully characterized from the analysis of UVLS which provided the size and refractive index (RI) of individual beads. We believe that the main application of UVLS relates to the analysis of microparticles in a serum, in particular in the analysis of chylomicrons (CMs). We have demonstrated the performance of the UVLS SFC in the analysis of CMs of a donor. The RI versus size scatterplot of CMs was successfully retrieved from the analysis. The current set-up of the SFC has allowed us to characterize individual CMs starting from the size of 160 nm that provides determination of the CM concentration in a serum with flow cytometry. This feature of the UVLS should help with the analysis of lipid metabolism measuring RI and size map evolution after lipase action.

KW - inverse problem

KW - light scattering

KW - scanning flow cytometry

KW - Particle Size

KW - Flow Cytometry

KW - Ultraviolet Rays

KW - Lipid Metabolism

KW - Scattering, Radiation

KW - Cell-Derived Microparticles

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85163119709&origin=inward&txGid=148a92982d57f39ad95089b1a5abbe4d

UR - https://www.mendeley.com/catalogue/3a18eedb-427a-3a68-8577-c9f622a0890f/

U2 - 10.1002/cyto.a.24769

DO - 10.1002/cyto.a.24769

M3 - Article

C2 - 37306103

VL - 103

SP - 736

EP - 743

JO - Cytometry. Part A : the journal of the International Society for Analytical Cytology

JF - Cytometry. Part A : the journal of the International Society for Analytical Cytology

SN - 1552-4922

IS - 9

ER -