DESIGN OF OPTICALLY ENCODED MICROSPHERES OF DIFFERENT DIAMETERS FOR MULTIPARAMETRIC FLOW CYTOMETRY
https://doi.org/10.56304/S2079562925010233
EDN: VIQHIN
Abstract
The development of innovative fluorescence detection methods based on optically encoded microspheres of different colors and sizes is an important direction of multiparameter detection and diagnostics of a wide range of diseases. Thus, the xMAP technology of the American company Luminex is based on the use of polystyrene microspheres colored with two or three organic fluorophores in different ratios, each of which has unique spectral characteristics. Despite the ability to detect up to 80 proteins or DNA sequences in a single test system, the need to use special equipment only from this company for multiplex analysis and the characteristic disadvantages of organic fluorophores (large Stokes shift, photo-bleaching under laser excitation) limit the application of this technology. The objectives of the present study consisted of the design and fabrication of microspheres encoded with semiconducting quantum dots. Carboxylated melamine-formaldehyde microspheres of two sizes were optically encoded with quantum dots of two colors immobilized on the surface of the microspheres by layer-by-layer adsorption of oppositely charged polyelectrolytes. As a result, six populations of microspheres with different sizes and/or unique optical codes were obtained with stability and homogeneity in aqueous solutions for a long time. Analysis of the obtained microspheres by dynamic light scattering, epifluorescence microscopy and flow cytometry methods showed their suitability for multiparameter analysis. At the same time, the use of quantum dots for optical encoding made it possible to exclude photodegradation of the signal and provided the possibility of excitation of all populations by the same wavelength of radiation with effective separation of signals from microspheres in different channels of a standard flow cytometer.
About the Authors
M. D. Sokolov
National Research Nuclear University MEPhI (Moscow Engineering Physics Institute)
Russian Federation
Russian Federation
E. S. Gerasimovich
National Research Nuclear University MEPhI (Moscow Engineering Physics Institute)
Russian Federation
Russian Federation
I. R. Nabiev
National Research Nuclear University MEPhI (Moscow Engineering Physics Institute);
Université de Reims Champagne-Ardenne
Russian Federation
Russian Federation
A. V. Sukhanova
Université de Reims Champagne-Ardenne
Russian Federation
Russian Federation
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Review
For citations:
Sokolov M.D., Gerasimovich E.S., Nabiev I.R., Sukhanova A.V. DESIGN OF OPTICALLY ENCODED MICROSPHERES OF DIFFERENT DIAMETERS FOR MULTIPARAMETRIC FLOW CYTOMETRY. Nuclear Physics and Engineering. 2025;16(5):721-727. (In Russ.) https://doi.org/10.56304/S2079562925010233. EDN: VIQHIN
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