Super‐resolved calibration‐free flow cytometric characterization of platelets and cell‐derived microparticles in platelet‐rich plasma

Konokhova, Anastasiya I.; Chernova, Daria N.; Moskalensky, Alexander E.; Strokotov, Dmitry I.; Yurkin, Maxim A.; Chernyshev, Andrei V.; Maltsev, Valeri P.

doi:10.1002/cyto.a.22621

Cited by 30 publications

(40 citation statements)

References 48 publications

(65 reference statements)

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“…Submicron particles are defined as particles with sizes below 1 µm in diameter, while nanoparticles are generally defined as particles that have a diameter between 1 and 100 nm in at least one dimension. Currently, a number of microscopic and biophysical techniques are used to characterize submicron and nanoparticles . Unfortunately, there is not a universal “gold” standard test that can be used to quantify and characterize submicron and nanoparticles in suspension.…”

Section: Detection Range Of Particle Sizesmentioning

confidence: 99%

Characterization, detection, and counting of metal nanoparticles using flow cytometry

2015

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There is a need to accurately detect, characterize, and quantify nanoparticles in suspensions. This study helps to understand the complex interactions between similar types of nanoparticles. Before initiating a study of metal nanoparticles, five submicron PS beads with sizes between 200 nm and 1 mm were used to derive a reference scale that was useful in evaluating the flow cytometer for functionality, sensitivity, resolution, and reproducibility. Side scatter intensity (SSC) from metal nanoparticles was obtained simultaneously from 405 nm and 488 nm lasers. The 405 nm laser generally yielded histogram distributions with smaller CVs, less side scatter intensity, better separation indices between beads and decreased scatter differences between different sized particles compared with the 488 nm laser. Submicron particles must be diluted to 10 6 and 10 7 particles/mL before flow cytometer analysis to avoid coincidence counting artifacts. When particles were too concentrated the following occurred: swarm, electronic overload, coincidence counting, activation of doublet discrimination and rejection circuitry, increase of mean SSC histogram distributions, alterations of SSC and pulse width histogram shape, decrease and fluctuations in counting rate and decrease or elimination of particulate water noise and 1 mm reference bead. To insure that the concentrations were in the proper counting range, the nanoparticle samples were mixed with a known concentration of 1mm counting beads. Sequential dilutions of metal nanoparticles in a 1 mm counting bead suspension helped determine the diluted concentration needed for flow cytometer analysis. It was found that the original concentrated nanoparticle samples had to be diluted, between 1:10,000 and 1:100,000, before characterization by flow cytometry. The concentration of silver or gold nanoparticles in the undiluted sample were determined by comparing them with a known concentration (1.9 3 10 6 beads/mL) of 1 mm polystyrene reference beads. Published 2015 Wiley Periodicals Inc., on behalf of ISAC

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Section: Detection Range Of Particle Sizesmentioning

confidence: 99%

Characterization, detection, and counting of metal nanoparticles using flow cytometry

2015

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“…Specifically, light scattering profile (LSP) is measured in a wide range of scattering angles. This approach has been successfully applied to RBCs (26,27), as well as to other biological particles, including platelets, bacteria, blood microparticles, and milk fat globules (24,(28)(29)(30)(31). This approach has been successfully applied to RBCs (26,27), as well as to other biological particles, including platelets, bacteria, blood microparticles, and milk fat globules (24,(28)(29)(30)(31).…”

Section: Introductionmentioning

confidence: 99%

“…The core of the characterization method is the solution of the inverse light-scattering (ILS) problem, using a parametric shape model of the characterized particle (24,25). This approach has been successfully applied to RBCs (26,27), as well as to other biological particles, including platelets, bacteria, blood microparticles, and milk fat globules (24,(28)(29)(30)(31). Conceptually similar approach, based on LSP measurement combined with fitting theoretical LSPs, has been recently demonstrated for determination of RBC diameters in a microfluidic channel (32).…”

Section: Introductionmentioning

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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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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“…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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Super‐resolved calibration‐free flow cytometric characterization of platelets and cell‐derived microparticles in platelet‐rich plasma

Cited by 30 publications

References 48 publications

Characterization, detection, and counting of metal nanoparticles using flow cytometry

Characterization, detection, and counting of metal nanoparticles using flow cytometry

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

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

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