scholarly journals A Novel Rapid Method of Red Blood Cell and Platelet Permeabilization and Staining for Flow Cytometry Analysis

2019 ◽  
Vol 96 (5) ◽  
pp. 426-435 ◽  
Author(s):  
Sixtine Fauré ◽  
Andreas Van Agthoven ◽  
Denis Bernot ◽  
Alexandre Altié ◽  
Michel Grino ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cell ◽  
Rapid Method ◽  
Red Blood Cell ◽  
Flow Cytometry Analysis

Flow cytometry analysis of dual red blood cell populations after bone marrow transplantation

2008 ◽  
Vol 89 (4) ◽  
pp. 741-747 ◽  
Author(s):  
D. Blanchard ◽  
V. Bruneau ◽  
F. Germond-Arnoult ◽  
D. Bernard ◽  
A. Gourbil ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Bone Marrow Transplantation ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Marrow Transplantation ◽  
Cell Populations ◽  
Flow Cytometry Analysis

scholarly journals Enumeration of the Invasion Efficiency ofPlasmodium falciparumIn Vitro in Four Different Red Blood Cell Populations Using a Three‐Color Flow Cytometry‐Based Method

2019 ◽  
Vol 95 (7) ◽  
pp. 737-745 ◽  
Author(s):  
Sinmanus Vimonpatranon ◽  
Kesinee Chotivanich ◽  
Kasama Sukapirom ◽  
Sakaorat Lertjuthaporn ◽  
Ladawan Khowawisetsut ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Cell Populations ◽  
Color Flow

A preliminary comparison of flow cytometry and tube agglutination assays in detecting red blood cell-associated C3d

2003 ◽  
Vol 13 (1) ◽  
pp. 35-42 ◽  
Author(s):  
D. F. Stroncek ◽  
J. M. Njoroge ◽  
J. L. Procter ◽  
R. W. Childs ◽  
J. Miller
Keyword(s):  
Flow Cytometry ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Preliminary Comparison

Activated neutrophil-mediated sickle red blood cell adhesion to lung vascular endothelium: role of phosphatidylserine-exposed sickle red blood cells

2006 ◽  
Vol 291 (4) ◽  
pp. H1679-H1685 ◽  
Author(s):  
Johnson Haynes ◽  
Boniface Obiako ◽  
Judy A. King ◽  
Raymond B. Hester ◽  
Solomon Ofori-Acquah
Keyword(s):  
Flow Cytometry ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Vascular Endothelium ◽  
Pulmonary Circulation ◽  
Membrane Surface ◽  
Annexin V ◽  
Specific Marker ◽  
Double Labeling

Activated neutrophils (ANs) increase sickle red blood cell (SRBC) retention/adhesion in the pulmonary circulation. This study investigates the role of neutrophil activation and SRBC retention/adhesion in the pulmonary circulation through a mechanism that involves increasing phosphatidylserine (PS) exposure on the external membrane surface of the SRBCs (PS-exposed). With the use of flow cytometry, double-labeling studies were performed with a calcium-dependent phospholipid-binding protein, annexin V-fluorescein isothiocyanate fluorescence, and the erythroid-specific marker glycophorin A to assess for the percentage of PS-exposed normal and SRBCs at baseline and after coincubation with ANs. Additional studies were performed that assessed retention/adhesion of SRBCs in the isolated rat lung using51Cr-labeled SRBC alone, SRBC + AN, SRBC + AN + zileuton, and SRBC + AN + annexin V. Specific activities of lung and perfusate were measured, and the number of retained SRBCs per gram lung was calculated. Flow cytometry demonstrated that ANs increased the percentage of PS-exposed normal and SRBCs. The 5-lipoxygenase inhibitor zileuton attenuated AN-mediated increases in PS-exposed SRBCs and decreased SRBC retention/adherence in the lung on histological sections. Similarly, in the isolated perfused lung and in histological lung sections, retention/adherence of SRBCs cloaked with annexin V was attenuated in the presence of ANs. We conclude that ANs enhance the adhesion of SRBCs to vascular endothelium by increasing red blood cell membrane externalization of PS. Zileuton attenuation of AN-mediated SRBC PS externalization suggests that a 5-lipoxygenase product(s), secreted by the AN, plays a vital role in altering the adhesive properties of PS-exposed SRBCs to vascular endothelium.

scholarly journals Rapid Cl−/HCO3−exchange kinetics of AE1 in HEK293 cells and hereditary stomatocytosis red blood cells

2013 ◽  
Vol 305 (6) ◽  
pp. C654-C662 ◽  
Author(s):  
Etienne Frumence ◽  
Sandrine Genetet ◽  
Pierre Ripoche ◽  
Achille Iolascon ◽  
Immacolata Andolfo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cell ◽  
Red Blood Cells ◽  
Anion Exchange ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Hek293 Cells ◽  
Single Amino Acid ◽  
Stopped Flow ◽  
Wild Type

Anion exchanger 1 (AE1) or band 3 is a membrane protein responsible for the rapid exchange of chloride for bicarbonate across the red blood cell membrane. Nine mutations leading to single amino-acid substitutions in the transmembrane domain of AE1 are associated with dominant hereditary stomatocytosis, monovalent cation leaks, and reduced anion exchange activity. We set up a stopped-flow spectrofluorometry assay coupled with flow cytometry to investigate the anion transport and membrane expression characteristics of wild-type recombinant AE1 in HEK293 cells, using an inducible expression system. Likewise, study of three stomatocytosis-associated mutations (R730C, E758K, and G796R), allowed the validation of our method. Measurement of the rapid and specific chloride/bicarbonate exchange by surface expressed AE1 showed that E758K mutant was fully active compared with wild-type (WT) AE1, whereas R730C and G796R mutants were inactive, reinforcing previously reported data on other experimental models. Stopped-flow analysis of AE1 transport activity in red blood cell ghost preparations revealed a 50% reduction of G796R compared with WT AE1 corresponding to a loss of function of the G796R mutated protein, in accordance with the heterozygous status of the AE1 variant patients. In conclusion, stopped-flow led to measurement of rapid transport kinetics using the natural substrate for AE1 and, conjugated with flow cytometry, allowed a reliable correlation of chloride/bicarbonate exchange to surface expression of AE1, both in recombinant cells and ghosts and therefore a fine comparison of function between different stomatocytosis samples. This technical approach thus provides significant improvements in anion exchange analysis in red blood cells.

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