Fluorescence‐free flow cytometry for measurement of shape index distribution of resting, partially activated, and fully activated platelets

Litvinenko, Alena L.; Moskalensky, Alexander E.; Karmadonova, N.A.; Nekrasov, Vyacheslav M.; Strokotov, Dmitry I.; Konokhova, Anastasiya I.; Yurkin, Maxim A.; Pokushalov, Evgeny; Чернышев, А. В.; Maltsev, Valeri P.

doi:10.1002/cyto.a.23003

Cited by 22 publications

(18 citation statements)

References 47 publications

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“…Moreover, it can be easily combined with consumable-free analysis of blood platelets (37), thus covering the two most representative cells in blood. Moreover, it can be easily combined with consumable-free analysis of blood platelets (37), thus covering the two most representative cells in blood.…”

Section: Technical Notementioning

confidence: 99%

“…To conclude, absence of consumables and fast analysis time make the proposed method perfect for routine clinical diagnostics, significantly increasing its descriptiveness and sensitivity to pathologies interfering with RBC characteristics. Moreover, it can be easily combined with consumable-free analysis of blood platelets (37), thus covering the two most representative cells in blood.…”

Section: Technical Notementioning

confidence: 99%

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Advanced consumable‐free morphological analysis of intact red blood cells by a compact scanning flow cytometer

et al. 2017

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Whereas modern automated blood cell analyzers measure the volume of individual red blood cells (RBCs), leading to four RBC indices (mean corpuscular volume, MCV; mean corpuscular hemoglobin, MCH; mean corpuscular hemoglobin concentration, MCHC; red cell distribution width, and RDW), the RBC shape has not been assessed by clinical screening tools. We applied the scanning flow cytometer (SFC) for complete characterization of intact RBC morphology in terms of diameter, maximal and minimal thicknesses, volume, surface area, sphericity index, spontaneous curvature, hemoglobin concentration, and content. The above-mentioned individual RBC characteristics were measured without fluorescent markers and other chemicals by a SFC equipped only with 660 nm laser for RBC illumination and single detector for measurement of angle-resolved light scattering. The distributions over all RBC characteristics were constructed and processed statistically to form the novel 31 RBC indices for 22 donor samples. Our results confirm the possibility of precise, label-free, enhanced morphological analysis of individual intact RBCs with compact single-detector flow cytometer. Detailed characterization of RBCs with high statistics and precision can be used to increase the value of screening examinations and to reveal pathologies accompanied by abnormality of RBC shape. © 2017 International Society for Advancement of Cytometry.

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Section: Technical Notementioning

confidence: 99%

Section: Technical Notementioning

confidence: 99%

Advanced consumable‐free morphological analysis of intact red blood cells by a compact scanning flow cytometer

et al. 2017

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“…Note also that the presented model of activated platelet not only possess one or two additional morphological parameter in comparison with models of resting platelets, but also lacks axial symmetry, which brings about the influence of Euler angle γ on the light-scattering patterns. These three additional parameters complicate the solution of the inverse light scattering problem and hinder the incorporation of the presented model into the existing algorithms [ 9 , 10 ]. Further theoretical and experimental research concerning optical properties of blood platelets is needed to extract new information and develop novel diagnostic techniques.…”

Section: Methodsmentioning

confidence: 99%

Method for the simulation of blood platelet shape and its evolution during activation

et al. 2018

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We present a simple physically based quantitative model of blood platelet shape and its evolution during agonist-induced activation. The model is based on the consideration of two major cytoskeletal elements: the marginal band of microtubules and the submembrane cortex. Mathematically, we consider the problem of minimization of surface area constrained to confine the marginal band and a certain cellular volume. For resting platelets, the marginal band appears as a peripheral ring, allowing for the analytical solution of the minimization problem. Upon activation, the marginal band coils out of plane and forms 3D convoluted structure. We show that its shape is well approximated by an overcurved circle, a mathematical concept of closed curve with constant excessive curvature. Possible mechanisms leading to such marginal band coiling are discussed, resulting in simple parametric expression for the marginal band shape during platelet activation. The excessive curvature of marginal band is a convenient state variable which tracks the progress of activation. The cell surface is determined using numerical optimization. The shapes are strictly mathematically defined by only three parameters and show good agreement with literature data. They can be utilized in simulation of platelets interaction with different physical fields, e.g. for the description of hydrodynamic and mechanical properties of platelets, leading to better understanding of platelets margination and adhesion and thrombus formation in blood flow. It would also facilitate precise characterization of platelets in clinical diagnosis, where a novel optical model is needed for the correct solution of inverse light-scattering problem.

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“…We used a scanning flow cytometer (SFC), described in detail elsewhere (37)(38)(39)(40)(41). The SFC was fabricated by Cytonova LLC (Novosibirsk, Russia).…”

Section: Instrumentationmentioning

confidence: 99%

Proposed Dynamics of CDB3 Activation in Human Erythrocytes by Nifedipine Studied with Scanning Flow Cytometry

et al. 2019

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Nifedipine is calcium channels and pumps blocker widely used in medicine. However, mechanisms of nifedipine action in blood are not clear. In particular, the influence of nifedipine on erythrocytes is far from completely understood. In this work, applying scanning flow cytometry, we observed experimentally for the first time the dynamics behind a significant increase of HCO 3 − /Cl − transmembrane exchange rate of CDB3 (main anion exchanger, AE1, Band 3, SLC4A1) of human erythrocytes in the presence of nifedipine in blood. It was found that the rate of CDB3 activation is not limited by the rate of nifedipine binding and/or Ca 2+ transport. In order to explain the experimental data, we suggested a kinetic model assuming that the rate of CDB3 activation is limited by the dynamics of the balance between two intracellular processes (1) the activation of CDB3 limited by its interaction with intracellular Ca 2+ , and (2) the spontaneous deactivation of CDB3. Thus the use of scanning flow cytometry allowed to clarify quantitatively the molecular kinetic mechanism of nifedipine action on human erythrocytes. In particular, the efficiency (~30) and rates of activation (~0.3 min −1 ) and deactivation (~10 −3 min −1 ) of CDB3 in human erythrocytes was evaluated for two donors.Nifedipine is widely used in medicine under the name of Adalat and other drugs. One common usage is the treatment of cardiovascular pathology such as atherosclerosis, coronary heart disease, hypertension, cerebral ischemia, and others (1). Other uses include painful spasms of the esophagus (2,3), Raynaud's phenomenon (4), tocolytic therapy (5). However, the effect of the treatment by nifedipine is not always positive (6). In particular, it is not clear (7) the influence of nifedipine on the rate of HCO 3− /Cl − transmembrane CDB3 protein (AE1, Band 3), which plays an important role in bicarbonate metabolism (8). Overall, a more complete understanding of nifedipine action in blood is needed.Nifedipine belongs to the subclass of dihydropyridines (DHPs), which are known as inhibitors of Ca 2+ transport across the erythrocyte membrane in both pathways: (1) inward leak through Ca 2+ entry channels (9-11), and (2) extrusion by plasma membrane Ca 2+ pumps (PMCA) (12,13). It was reported (13) that about 50% inhibition of PMCA was obtained at extracellular nifedipine concentration of 300 μM. Interestingly, the same extracellular concentration of 300 μM nifedipine was reported (14) to cause less (about 30%) inhibition of Ca 2+ entry channels of erythrocyte membrane. Stronger inhibition of PMCA than entry channels leads to the accumulation of intracellular Ca 2+ in erythrocytes in the presence of extracellular

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Fluorescence‐free flow cytometry for measurement of shape index distribution of resting, partially activated, and fully activated platelets

Cited by 22 publications

References 47 publications

Advanced consumable‐free morphological analysis of intact red blood cells by a compact scanning flow cytometer

Advanced consumable‐free morphological analysis of intact red blood cells by a compact scanning flow cytometer

Method for the simulation of blood platelet shape and its evolution during activation

Proposed Dynamics of CDB3 Activation in Human Erythrocytes by Nifedipine Studied with Scanning Flow Cytometry

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