scholarly journals Noninvasive imaging of flowing blood cells using label-free spectrally encoded flow cytometry

2012 ◽  
Vol 3 (6) ◽  
pp. 1455 ◽  
Author(s):  
Lior Golan ◽  
Daniella Yeheskely-Hayon ◽  
Limor Minai ◽  
Eldad J Dann ◽  
Dvir Yelin
Keyword(s):  
Flow Cytometry ◽  
Blood Cells ◽  
Noninvasive Imaging ◽  
Label Free ◽  
Flowing Blood ◽  
Spectrally Encoded

scholarly journals Photoacoustic Flow Cytometry for Single Sickle Cell DetectionIn VitroandIn Vivo

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Chengzhong Cai ◽  
Dmitry A. Nedosekin ◽  
Yulian A. Menyaev ◽  
Mustafa Sarimollaoglu ◽  
Mikhail A. Proskurnin ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Red Blood Cells ◽  
Real Time ◽  
Sickle Cell ◽  
Blood Cells ◽  
Label Free ◽  
Linear Mode ◽  
Signal Fluctuation

Control of sickle cell disease (SCD) stage and treatment efficiency are still time-consuming which makes well-timed prevention of SCD crisis difficult. We show here thatin vivophotoacoustic (PA) flow cytometry (PAFC) has a potential for real-time monitoring of circulating sickled cells in mouse model.In vivodata were verified byin vitroPAFC and photothermal (PT) and PA spectral imaging of sickle red blood cells (sRBCs) expressing SCD-associated hemoglobin (HbS) compared to normal red blood cells (nRBCs). We discovered that PT and PA signal amplitudes from sRBCs in linear mode were 2–4-fold lower than those from nRBCs. PT and PA imaging revealed more profound spatial hemoglobin heterogeneity in sRBCs than in nRBCs, which can be associated with the presence of HbS clusters with high local absorption. This hypothesis was confirmed in nonlinear mode through nanobubble formation around overheated HbS clusters accompanied by spatially selective signal amplification. More profound differences in absorption of sRBCs than in nRBCs led to notable increase in PA signal fluctuation (fluctuation PAFC mode) as an indicator of SCD. The obtained data suggest that noninvasive label-free fluctuation PAFC has a potential for real-time enumeration of sRBCs bothin vitroandin vivo.

scholarly journals Multi-pronged approach to human mesenchymal stromal cells senescence quantification with a focus on label-free methods

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Weichao Zhai ◽  
Jerome Tan ◽  
Tobias Russell ◽  
Sixun Chen ◽  
Dennis McGonagle ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Preclinical Model ◽  
Label Free ◽  
Current Standard ◽  
Differentiation Potential ◽  
Human Mesenchymal Stromal Cells ◽  
Endogenous Fluorophores

AbstractHuman mesenchymal stromal cells (hMSCs) have demonstrated, in various preclinical settings, consistent ability in promoting tissue healing and improving outcomes in animal disease models. However, translation from the preclinical model into clinical practice has proven to be considerably more difficult. One key challenge being the inability to perform in situ assessment of the hMSCs in continuous culture, where the accumulation of the senescent cells impairs the culture’s viability, differentiation potential and ultimately leads to reduced therapeutic efficacies. Histochemical $$\upbeta $$ β -galactosidase staining is the current standard for measuring hMSC senescence, but this method is destructive and not label-free. In this study, we have investigated alternatives in quantification of hMSCs senescence, which included flow cytometry methods that are based on a combination of cell size measurements and fluorescence detection of SA-$$\upbeta $$ β -galactosidase activity using the fluorogenic substrate, C$${_{12}}$$ 12 FDG; and autofluorescence methods that measure fluorescence output from endogenous fluorophores including lipopigments. For identification of senescent cells in the hMSC batches produced, the non-destructive and label-free methods could be a better way forward as they involve minimum manipulations of the cells of interest, increasing the final output of the therapeutic-grade hMSC cultures. In this work, we have grown hMSC cultures over a period of 7 months and compared early and senescent hMSC passages using the advanced flow cytometry and autofluorescence methods, which were benchmarked with the current standard in $$\upbeta $$ β -galactosidase staining. Both the advanced methods demonstrated statistically significant values, (r = 0.76, p $$\le $$ ≤ 0.001 for the fluorogenic C$${_{12}}$$ 12 FDG method, and r = 0.72, p $$\le $$ ≤ 0.05 for the forward scatter method), and good fold difference ranges (1.120–4.436 for total autofluorescence mean and 1.082–6.362 for lipopigment autofluorescence mean) between early and senescent passage hMSCs. Our autofluroescence imaging and spectra decomposition platform offers additional benefit in label-free characterisation of senescent hMSC cells and could be further developed for adoption for future in situ cellular senescence evaluation by the cell manufacturers.

Role of erythrocytes and platelets in the hypercoagulable status in polycythemia vera through phosphatidylserine exposure and microparticle generation

2013 ◽  
Vol 109 (06) ◽  
pp. 1025-1032 ◽  
Author(s):  
Chunyan Gao ◽  
Xue Yang ◽  
Jianan Li ◽  
Wei Wang ◽  
Jinxiao Hou ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cells ◽  
Significant Inhibition ◽  
Polycythaemia Vera ◽  
Factor V ◽  
Healthy Controls ◽  
Clotting Time ◽  
Thrombotic Risk ◽  
Few Data

SummaryThe development of thrombosis in polycythaemia vera (PV) involves multifactorial processes including pathological activation of blood cells. Release of microparticles (MPs) by activated cells in diseases is associated with thrombotic risk, but relatively few data are available in PV. The aim of the present study was to investigate the increase in MP release and exposure of phosphatidylserine (PS) on the outer membrane of MP-origin cells in patients with PV, and to analyse their procoagulant activity (PCA). PS-positive MPs and cells were detected by flow cytometry, while PCA was assessed with clotting time and purified coagulation complex assays. We found that PV patients had elevated circulating lactadherin+ MPs, which mostly originating from erythrocytes, platelets, granulocytes, and endothelial cells, as well as increased PS exposing erythrocytes/platelets as compared to secondary polycythaemia patients or healthy controls. These PS-bearing MPs and cells were highly procoagulant. Moreover, lactadherin competed factor V and VIII to PS and inhibited about 90% of the detected PCA in a dose-response manner while anti-TF antibody did no significant inhibition. Treatment with hydroxyurea is associated with a decrease in PS exposure and lactadherin+ MP release of erythrocytes/platelets. Our data demonstrate that PV patients are characterised by increased circulating procoagulant MPs and PS exposing erythrocytes/platelets, which could contribute to the hypercoagulable state in these patients.

Deep Learning-enabled Holographic Imaging Flow-Cytometry for Label-Free Detection of Giardia Lamblia in Water Samples

2021 ◽  
Author(s):  
Zoltán Göröcs ◽  
David Baum ◽  
Fang Song ◽  
Kevin de Haan ◽  
Hatice Ceylan Koydemir ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Deep Learning ◽  
Water Samples ◽  
Giardia Lamblia ◽  
Label Free ◽  
Holographic Imaging ◽  
Imaging Flow Cytometry ◽  
Label Free Detection

scholarly journals Leukocytes in bovine virgin mammary gland: flow cytometry imaging during development and resolution of induced influx

2001 ◽  
Vol 46 (No. 7–8) ◽  
pp. 190-198
Author(s):  
Z. Sládek ◽  
D. Ryšánek ◽  
M. Faldyna
Keyword(s):  
Flow Cytometry ◽  
Mammary Gland ◽  
Blood Cells ◽  
Cell Types ◽  
Mammary Glands ◽  
Mononuclear Phagocytes ◽  
Peripheral Blood Cells ◽  
Leukocyte Influx ◽  
Separate Region ◽  
Blood Leukocyte

Distribution of leukocyte types present in virgin bovine mammary glands was analysed in dot plots obtained by flow cytometry (FACS) of samples collected from 10 non-pregnant heifers after induction of leukocyte influx. Changes of percentage of leukocyte types during development and resolution of induced influx in comparison with blood leukocyte pattern allow identification of these cell types on FACS dot plot. The positions of mammary gland granulocyte and lymphocyte regions were identical with those of the corresponding peripheral blood cells. Two basic morphologically distinct types occupying separate regions in dot plots were observed in the population of mononuclear phagocytes (MoP): non-vacuolised monocyte-like macrophages (MoMAC) and vacuolised macrophages (MAC). Influx resolution was characterised by a marked shift of the MoMAC region towards that of MAC recognisable in dot plots by a separate region of intermediate MoP forms. The study provides a pattern of dynamics of percentages of mammary gland leukocyte types during influx development and resolution as imaged by FACS.

Microparticles in stored red blood cells: an approach using flow cytometry and proteomic tools

Vox Sanguinis ◽  
2008 ◽  
Vol 95 (4) ◽  
pp. 288-297 ◽  
Author(s):  
O. Rubin ◽  
D. Crettaz ◽  
G. Canellini ◽  
J.-D. Tissot ◽  
N. Lion
Keyword(s):  
Flow Cytometry ◽  
Red Blood Cells ◽  
Blood Cells

scholarly journals Simultaneous acoustic and photoacoustic microfluidic flow cytometry for label-free analysis

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Vaskar Gnyawali ◽  
Eric M. Strohm ◽  
Jun-Zhi Wang ◽  
Scott S. H. Tsai ◽  
Michael C. Kolios
Keyword(s):  
Flow Cytometry ◽  
Label Free ◽  
Microfluidic Flow

Classification of blood cells and tumor cells using label-free ultrasound and photoacoustics

2015 ◽  
Vol 87 (8) ◽  
pp. 741-749 ◽  
Author(s):  
Eric M. Strohm ◽  
Michael C. Kolios
Keyword(s):  
Tumor Cells ◽  
Blood Cells ◽  
Label Free

7.2 Hematology including Flow Cytometry of Blood Cells

2015 ◽  
Author(s):  
Giuseppe Banfi ◽  
Walter G. Guder ◽  
Sheshadri Narayanan
Keyword(s):  
Flow Cytometry ◽  
Blood Cells
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