Light-scattering flow cytometry for identification and characterization of blood microparticles

Konokhova, Anastasiya I.; Yurkin, Maxim A.; Moskalensky, Alexander E.; Chernyshev, Andrei V.; Tsvetovskaya, Galina A.; Chikova, Elena D.; Maltsev, Valeri P.

doi:10.1117/1.jbo.17.5.057006

Cited by 50 publications

(65 citation statements)

References 19 publications

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“…The activation was recorded by flow cytometry, which allows estimating the size of the cells (the change of front light scattering) and the granularity of their cytoplasm (the degree of lateral light scattering) (18). Since the activation of platelets is accompanied by changes in their shape and granulation, the change of front and lateral light scattering showed the level of platelet activation (Figure 5).…”

Section: Resultsmentioning

confidence: 99%

Antiplatelet and anti-proliferative action of disintegrin from Echis multisquamatis snake venom

et al. 2017

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AimTo purify the platelet aggregation inhibitor from Echis multisquamatis snake venom (PAIEM) and characterize its effect on platelet aggregation and HeLa cell proliferation.MethodsSodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) were used for PAIEM identification. Platelet aggregation in the presence of PAIEM was studied on aggregometer Solar-AP2110. The changes of shape and granularity of platelets in the presence of PAIEM were studied on flow cytometer COULTER EPICS XL, and degranulation of platelets was estimated using spectrofluorimetry. Indirect enzyme-linked immunosorbent assay was used for the determination of target of PAIEM on platelet surface. An assay based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was used to evaluate the effect of PAIEM on the proliferation of HeLa cells in cell culture.ResultsThe molecular weight of the protein purified from Echis multisquamatis venom was 14.9 kDa. Half-maximal inhibitory concentration (IC50) of PAIEM needed to inhibit adenosine diphosphate (ADP)-induced platelet aggregation was 7 μM. PAIEM did not affect thrombin- or ADP-induced platelet activation, but it did prevent binding of the anti-IIb antibody to glycoprotein IIb/IIIa (GPIIbIIIa)-receptor of adhered platelets and inhibited the viability of HeLa cells by 54%.ConclusionAs a member of the disintegrin family, PAIEM inhibited platelet aggregation and cell proliferation possibly by blocking integrin-mediated interactions. However, it did not impair cellular signaling causing any changes in platelet shape and granularity and did not affect ADP-induced platelet degranulation. This disintegrin was shown to be a potent inhibitor of integrin-mediated cellular interactions including platelet aggregation or cancer cell proliferation.

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Section: Resultsmentioning

confidence: 99%

Antiplatelet and anti-proliferative action of disintegrin from Echis multisquamatis snake venom

et al. 2017

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“…3). Recently, the refractive index range of vesicles was estimated to range between 1.36 and 1.45 [22], which would result in a size estimate of 190-1040 nm corresponding to a signal of 10 À6 mW in Fig. 3.…”

Section: Flow Cytometrymentioning

confidence: 99%

Innovation in detection of microparticles and exosomes

Pol

Coumans

Varga

et al. 2013

Journal of Thrombosis and Haemostasis

224

194

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Summary. Cell-derived or extracellular vesicles, including microparticles and exosomes, are abundantly present in body fluids such as blood. Although such vesicles have gained strong clinical and scientific interest, their detection is difficult because many vesicles are extremely small with a diameter of less than 100 nm, and, moreover, these vesicles have a low refractive index and are heterogeneous in both size and composition. In this review, we focus on the relatively high throughput detection of vesicles in suspension by flow cytometry, resistive pulse sensing, and nanoparticle tracking analysis, and we will discuss their applicability and limitations. Finally, we discuss four methods that are not commercially available: Raman microspectroscopy, micro nuclear magnetic resonance, small-angle X-ray scattering (SAXS), and anomalous SAXS. These methods are currently being explored to study vesicles and are likely to offer novel information for future developments.

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“…2 The characteristics of particles can be accurately determined when the particle geometry is relatively simple. [5][6][7] Scattering of light by particles with complicated geometry is more difficult to interpret, mainly due to a larger number of characteristics. [5][6][7] Scattering of light by particles with complicated geometry is more difficult to interpret, mainly due to a larger number of characteristics.…”

Section: Introductionmentioning

confidence: 99%

Additivity of light-scattering patterns of aggregated biological particles

et al. 2014

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The paper is focused on light scattering by aggregates of optically soft particles with a size larger than the wavelength, in particular, blood platelets. We conducted a systematic simulation of light scattering by dimers and larger aggregates of blood platelets, each modeled as oblate spheroids, using the discrete dipole approximation. Two-dimensional (2-D) light scattering patterns (LSPs) and internal fields showed that the multiple scattering between constituent particles can be neglected. Additionally, we derived conditions of the scattering angle and orientation of the dimer, under which the averaging of the 2-D LSPs over the azimuthal scattering angle washes out interference in the far field, resulting in averaged LSPs of the aggregate being equal to the sum of that for its constituents. We verified theoretical conclusions using the averaged LSPs of blood platelets measured with the scanning flow cytometer (SFC). Moreover, we obtained similar results for a model system of aggregates of polystyrene beads, studied both experimentally and theoretically. Finally, we discussed the potential of discriminating platelet aggregates from monomers using the SFC.

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Light-scattering flow cytometry for identification and characterization of blood microparticles

Cited by 50 publications

References 19 publications

Antiplatelet and anti-proliferative action of disintegrin from Echis multisquamatis snake venom

Antiplatelet and anti-proliferative action of disintegrin from Echis multisquamatis snake venom

Innovation in detection of microparticles and exosomes

Additivity of light-scattering patterns of aggregated biological particles

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