scholarly journals A new microparticle size calibration standard for use in measuring smaller microparticles using a new flow cytometer

2011 ◽  
Vol 9 (6) ◽  
pp. 1216-1224 ◽  
Author(s):  
W. L. CHANDLER ◽  
W. YEUNG ◽  
J. F. TAIT
1991 ◽  
Vol 66 (03) ◽  
pp. 350-354 ◽  
Author(s):  
Rob Fijnheer ◽  
Christa H E Homburg ◽  
Berend Hooibrink ◽  
Martine N Boomgaard ◽  
Dirk de Korte ◽  
...  

SummaryThrombin-induced changes in cytosolic free Ca2+ ([Ca2+]i) were studied in human platelets that had been stored for up to 6 days. Changes in [Ca2+]i were measured with Indo-1-loaded platelets and quantitated with two different methods: (i) measurement of the changes in total fluorescence; (ii) measurement of the [Ca2+]i changes in individual platelets in a flow cytometer, allowing the detection of non-responding platelets. The maximal concentration of [Ca2+]i after stimulation with 0.5 U of thrombin/ml decreased from 544 ± 58 nM (mean ± SEM, n = 6) on day 0, to 276 ± 9 nM on day 3 and to 203 ± 23 nM on day 6. The percentage of platelets responding to 0.5 U of thrombin/ml declined from 90 ± 2% on day 0 to 72 ± 4% on day 3, and to 47 ± 8% on day 6. Nevertheless, also the responding platelets showed a decreased rise in [Ca2+]i.The study shows that during platelet storage a decrease in the rise in [Ca2+]i upon thrombin stimulation occurs. This decrease is partly due to the formation of a subpopulation of platelets that is completely unresponsive and partly due to a decreased responsiveness in the remainder of the platelets; it is not due to a gradual decline in [Ca2+]i rise in all platelets. This phenomenon provides new insight in the functional defect of stored platelets.


Vsyo o myase ◽  
2020 ◽  
pp. 56-60
Author(s):  
Zayko E.V. ◽  
◽  
Kuznetsova O.A. ◽  
Bataeva D.S. ◽  
Grudistova M.A. ◽  
...  

The problem of the uncontrolled use of antibiotics currently remains unresolved. Step-by-step monitoring of meat using modern methods will reduce the risk of using contaminated meat raw materials for food production. Qualitative monitoring will identify samples containing residual antimicrobial substances. The use of methods for identifying groups of antibiotics will help narrow the search for antibiotics by expensive chromatographic methods. A study was carried out of beef, pork and poultry meat, which is used in meat processing plants in the production of raw smoked sausages, using two methods. At the first stage, using a qualitative microbiological method, the raw meat was evaluated for the presence of antimicrobial substances, then their group was determined using a NovoCyte flow cytometer. According to the results of a study on a flow cytometer, it was found that out of 10 groups of antibiotics that can be determined by the tested method, the group of lincosamides was not found in all meat samples. The most common groups of chemotherapy drugs in pork were sulfonamides – 29.6 %, tetracycline group – 18.5 % and beta-lactams – 14.8 %, and in beef aminoglycosides – 36.7 %, phenicols – 30 % and beta-lactams – 13.3 %. In poultry meat samples, the most common were sulfonamides – 23.2 %, fenicols – 23.2 %, and beta-lactams – 16 %. Five groups of antibiotics were found in all studied types of meat: fenicols, β-lactams, macrolides, polypeptide antibiotics, and quinolones. This indicates their widespread use in animal husbandry and poultry farming.


2021 ◽  
Vol 10 (2) ◽  
pp. 319
Author(s):  
Hee Cheol Yang ◽  
Won Jong Rhee

Because cancers are heterogeneous, it is evident that multiplexed detection is required to achieve disease diagnosis with high accuracy and specificity. Extracellular vesicles (EVs) have been a subject of great interest as sources of novel biomarkers for cancer liquid biopsy. However, EVs are nano-sized particles that are difficult to handle; thus, it is necessary to develop a method that enables efficient and straightforward EV biomarker detection. In the present study, we developed a method for single step in situ detection of EV surface proteins and inner miRNAs simultaneously using a flow cytometer. CD63 antibody and molecular beacon-21 were investigated for multiplexed biomarker detection in normal and cancer EVs. A phospholipid-polymer-phospholipid conjugate was introduced to induce clustering of the EVs analyzed using nanoparticle tracking analysis, which enhanced the detection signals. As a result, the method could detect and distinguish cancer cell-derived EVs using a flow cytometer. Thus, single step in situ detection of multiple EV biomarkers using a flow cytometer can be applied as a simple, labor- and time-saving, non-invasive liquid biopsy for the diagnosis of various diseases, including cancer.


2021 ◽  
Vol 22 (15) ◽  
pp. 8256
Author(s):  
Adolfas K. Gaigalas ◽  
Yu-Zhong Zhang ◽  
Linhua Tian ◽  
Lili Wang

A stochastic model of the flow cytometer measurement process was developed to assess the nature of the observed coefficient of variation (CV%) of the mean fluorescence intensity (MFI) from a population of labeled microspheres (beads). Several sources of variability were considered: the total number of labels on a bead, the path through the laser beam, the optical absorption cross-section, the quantum yield, the numerical aperture of the collection optics, and the photoelectron conversion efficiency of the photomultiplier (PMT) cathode. The variation in the number of labels on a bead had the largest effect on the CV% of the MFI of the bead population. The variation in the path of the bead through the laser beam was minimized using flat-top lasers. The variability in the average optical properties of the labels was of minor importance for beads with sufficiently large number of labels. The application of the bead results to the measured CV% of labeled B cells indicated that the measured CV% was a reliable measure of the variability of antibodies bound per cell. With some modifications, the model can be extended to multicolor flow cytometers and to the study of CV% from cells with low fluorescence signal.


Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1320
Author(s):  
Kristýna Pekárková ◽  
Jakub Soukup ◽  
Marie Kostelanská ◽  
Jan Širc ◽  
Zbyněk Straňák ◽  
...  

Extracellular vesicles (EVs) from liquid biopsies are extensively analyzed by flow cytometry, a technology that is continuously evolving. Thresholding utilizing a violet 405 nm laser side scatter (VSSC) has recently been implemented. Here, we collected set of large EV (lEV) samples from cord blood, which we analyzed using a standard flow cytometer improved via a 405 nm laser side scatter. Samples were analyzed using two distinct thresholding methods—one based on VSSC, and one based on VSSC combined with fluorescence thresholding on stained phosphatidylserine. Through these thresholding methods, we compared lEVs from pre-term births and control cord blood. Double-labeled lEVs with platelet CD36+/CD41+, activated platelet CD41+/CD62P+ and endothelial CD31+/CD105+ antibodies were used. Apart from comparing the two groups together, we also correlated measured lEVs with the thresholding methods. We also correlated the results of this study with data analyzed in our previous study in which we used a conventional 488 nm laser SSC. We did not find any difference between the two cord blood groups. However, we found highly concurrent data via our correlation of the thresholding methods, with correlation coefficients ranging from 0.80 to 0.96 even though the numbers of detected lEVs differed between thresholding methods. In conclusion, our approaches to thresholding provided concurrent data and it seems that improving the cytometer with the use of a VSSC increases its sensitivity, despite not being particularly critical to the validity of flow cytometric studies that compare pathological and physiological conditions in liquid biopsies.


1999 ◽  
Vol 45 (10) ◽  
pp. 1879-1880 ◽  
Author(s):  
Andreas Lun ◽  
Reinhard Ziebig ◽  
Hannes Hammer ◽  
Uwe Otting ◽  
Guido Filler ◽  
...  

2005 ◽  
Vol 12 (12) ◽  
pp. 1416-1424 ◽  
Author(s):  
Kovit Pattanapanyasat ◽  
Surada Lerdwana ◽  
Egarit Noulsri ◽  
Thanyanan Chaowanachan ◽  
Punneeporn Wasinrapee ◽  
...  

ABSTRACT Use of the standard dual-platform flow cytometric method for determination of CD4+ T-lymphocyte counts, which needs both a flow cytometer (FCM) and hematological analyzer, would inevitably lead to increased variability. The development of new single-platform (SP) FCMs that provide direct CD4+ T-lymphocyte counts for improved assay precision and accuracy have recently attracted attention. This study evaluated one of those systems, CyFlowgreen (Partec), a single-parameter SP volumetric FCM. The performance of CyFlowgreen was compared with those of two reference standard SP microbead-based technologies of the three-color TruCOUNT tube with the FACScan FCM and a two-color FACSCount system (Becton Dickinson Biosciences). Absolute CD4+ and CD8+ T-lymphocyte counts in 200 human immunodeficiency virus type 1-seropositive blood specimens were determined. Statistical analysis for correlation and agreement were performed. A high correlation of absolute CD4 counts was shown when those obtained with CyFlowgreen were compared with those obtained with the bead-based three-color TruCOUNT system (R 2 = 0.96; mean bias, −69.1 cells/μl; 95% confidence interval [CI], −225.7 to +87.5 cells/μl) and the FACSCount system (R 2 = 0.97; mean bias, −40.0 cells/μl; 95% CI, −165.1 to +85.1 cells/μl). The correlation of the CD4+ T-lymphocyte counts obtained by the two bead-based systems was high (R 2 = 0.98). Interestingly, CyFlowgreen yielded CD4+ T-lymphocyte counts that were 21.8 and 7.2 cells/μl lower than those obtained with the TruCOUNT and the FACSCount systems, respectively, when CD4+ T-lymphocyte counts were <250 CD4+ T-lymphocyte counts/μl range or 17.3 and 5.8 cells/μl less, respectively, when CD4+ T-lymphocyte counts were <200 cells/μl. The single-parameter CyFlowgreen volumetric technology performed well in comparison with the performance of the standard SP bead-based FCM system. However, a multicenter comparative study is needed before this FCM machine is implemented in resource-limited settings.


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