Kinetic study of the initial stages of agglutination process with scanning flow cytometer

Ivan V. Surovtsev; Maxim A. Yurkin; Alexander N. Shvalov; Galina F. Sivolobova

doi:10.1117/12.477906

19 June 2003 Kinetic study of the initial stages of agglutination process with scanning flow cytometer

Ivan V. Surovtsev, Maxim A. Yurkin, Alexander N. Shvalov, Galina F. Sivolobova

Proceedings Volume 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues; (2003) https://doi.org/10.1117/12.477906
Event: Biomedical Optics, 2003, San Jose, CA, United States

Abstract

The main disadvantage in ordinary agglutination immunoassay is difficulties to distinguish between specific and non-specific particle aggregation. We proposed to use Scanning Flow Cytometry to the kinetic study of the initial stages of agglutination process. The main advantage of the Scanning Flow Cytometry is a possibility to measure angular dependency of the light scattered by a single particle, an indicatrix. The most promising field for application of the indicatrix technology is a characterization of non-spherical particles. Validity of proposed method was verified by simultaneous measurements of the light scattering and fluorescence signal. We used Wentzel-Kramers-Brillouin approximation to simulate light scattering from two glued spheres and to explain the results obtained from measured indicatrices. To show an applicability of the proposed technique, the kinetic experiments were performed on latex particle covered with BSA (diameter 1.8 μm). Kinetics of dimer fraction growth initiated by mixing BSA-covered latex particles with anti-BSA immuoglobulins IgG was studied. In order to evaluate kinetic rate constant simple kinetic model involved only dimer growth reaction was applied for data treatment. Two kinetic rate constant for dimer fraction growth k_B=2.88•10^-12 cm³s^-1 and k_A=0.85•10^-12 cm³s^-1 were evaluated for two samples with the same origin but with different prehistory.

Citation Download Citation

Ivan V. Surovtsev, Maxim A. Yurkin, Alexander N. Shvalov, and Galina F. Sivolobova "Kinetic study of the initial stages of agglutination process with scanning flow cytometer", Proc. SPIE 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues, (19 June 2003); https://doi.org/10.1117/12.477906

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KEYWORDS

Particles

Light scattering

Optical spheres

Latex

Luminescence

Diffusion

Flow cytometry

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