scholarly journals Cell Surface Flow Cytometry Staining of Whole Blood v4 (protocols.io.babbiain)

protocols.io ◽  
2019 ◽  
Author(s):  
Sam Li
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Whole Blood ◽  
Surface Flow ◽  
Flow Cytometry Staining

Glycoconjugates on the surface of spores of the pathogenic fungus Phytophthora cinnamomi studied using fluorescence and electron microscopy and flow cytometry

1990 ◽  
Vol 36 (3) ◽  
pp. 183-192 ◽  
Author(s):  
A. R. Hardham ◽  
E. Suzaki
Keyword(s):  
Flow Cytometry ◽  
Plasma Membrane ◽  
Cell Surface ◽  
Phytophthora Cinnamomi ◽  
Pathogenic Fungus ◽  
Fluorescein Isothiocyanate ◽  
Pathogenic Fungi ◽  
Surface Flow ◽  
Soybean Agglutinin ◽  
Binding Material

Glycoconjugates on the surface of zoospores and cysts of the pathogenic fungus Phytophthora cinnamomi have been studied using fluorescein isothiocyanate labelled lectins for fluorescence microscopy and flow cytometry, and ferritin- and gold-labelled lectins for ultrastructural analysis. Of the five lectins used, only concanavalin A (ConA) binds to the surface of the zoospores, including the flagella and water expulsion vacuole. This suggests that of accessible saccharides, glucosyl or mannosyl residues predominate on the outer surface of the zoospore plasma membrane. Early in encystment, a system of flat disc-like cisternae, which underlie the zoospore plasma membrane, vesiculate. These and other small peripheral vesicles quickly disappear. After the induction of encystment, ConA is no longer localised close to the plasma membrane but binds to material loosely associated with the cell surface. Quantitative measurements by flow cytometry indicate that the ConA-binding material is gradually lost from the cell surface. The cyst wall is weakly labelled, but the site of germ tube emergence stains intensely. During the first 2 min after the induction of encystment, material that binds soybean agglutinin, Helix pommatia agglutinin, and peanut agglutinin appears on the surface of the fungal cells. The distribution of this material, rich in galactosyl or N-acetyl-D-galactosaminosyl residues, is initially patchy, but by 5 min the material evenly coats most of the cell surface. Labelling of zoospores in which intracellular sites are accessible indicates that the soybean agglutinin binding material is stored in vesicles that lie beneath the plasma membrane. Quantitation of soybean agglutinin labelling shows that maximum binding occurs 2–3 min after the induction of encystment. Key words: cell surface, flow cytometry, lectins, pathogenic fungi, Phytophthora cinnamomi.

scholarly journals Bicentric evaluation of stabilizing sampling tubes for assessment of monocyte HLA-DR expression in clinical samples

2021 ◽  
Author(s):  
guillaume monneret
Keyword(s):  
Flow Cytometry ◽  
Whole Blood ◽  
Human Leukocyte ◽  
Clinical Samples ◽  
University Hospitals ◽  
Short Delay ◽  
Leukocyte Antigen ◽  
Hla Dr ◽  
Healthy Donors ◽  
Flow Cytometry Staining

Background. Diminished expression of human leukocyte antigen DR on circulating monocytes (mHLA-DR), measured by standardized flow cytometry procedure, is a reliable indicator of immunosuppression in severely injured intensive care unit patients. As such, it is used as stratification criteria in clinical trials evaluating novel immunostimulating therapies. Pre-analytical constraints relative to the short delay between blood sampling and flow cytometry staining have nevertheless limited its use in multicentric studies. The objective of the present work was to compare mHLA-DR expression between whole blood samples simultaneously drawn in EDTA or Cyto-Chex BCT tubes. Methods. In 2 university hospitals, mHLA-DR was assessed in fresh whole blood from septic patients (n = 12) and healthy donors (n = 6) simultaneously sampled on EDTA and Cyto-Chex BCT tubes. Staining was performed immediately after sampling and after blood storage at room temperature. Results. We observed the remarkable stability of mHLA-DR results when blood was collected in Cyto-Chex BCT tubes (until 48-72 h). On baseline values, despite good correlation between tubes (r = 0.98, p< 0.001), mHLA-DR expression was systematically lower with Cyto-Chex BCT. Conclusion. The present reports confirms the great potential of Cyto-Chex BCT tubes to delay mHLA-DR staining in centers without rapid access to flow cytometry facilities. However, a 30 % gap exists between results obtained with EDTA and Cyto-Chex BCT tubes. As current thresholds for clinical decisions were obtained with EDTA samples, further studies are needed to confirm clinical thresholds with Cyto-Chex BCT tubes.

T-Cell Surface Markers in Human Peripheral Whole Blood Using Flow Cytometry

2008 ◽  
pp. 85-105 ◽  
Author(s):  
Manjula Reddy ◽  
Cuc Davis ◽  
Hugh Davis ◽  
Charles Pendley ◽  
Uma Prabhakar
Keyword(s):  
Flow Cytometry ◽  
T Cell ◽  
Cell Surface ◽  
Whole Blood ◽  
Surface Markers ◽  
Cell Surface Markers

Flow cytometry analysis of coxsackievirus B receptors expression in human CaCo-2 cells

2014 ◽  
Vol 9 (7) ◽  
pp. 699-707
Author(s):  
Samira Riabi ◽  
Rafik Harrath ◽  
Imed Gaâloul ◽  
Hind Hamzeh-Cognasse ◽  
Olivier Délezay ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Expression Patterns ◽  
Surface Expression ◽  
Surface Flow ◽  
Surface Proteins ◽  
Flow Cytometry Analysis ◽  
Coxsackie Adenovirus Receptor ◽  
Coxsackievirus B ◽  
Adenovirus Receptor

AbstractA subset of coxsackieviruses B (CV-B) is able to initiate intestinal infection via the attachment to two cell surface proteins, decayaccelerating factor (DAF) and coxsackie adenovirus receptor (CAR). The aim of the present study was to investigate the expression pattern of these receptors in the polarized CaCo-2 cell line using flow cytometry. The expression of CAR-specific mRNA and proteins was analyzed by reverse transcriptase polymerase chain reaction and western blotting, respectively. Flow cytometry analysis was used to study the surface expression patterns of CAR and DAF. CAR and DAF were well detected at the surface of CaCo-2 cells by flow cytometry. Despite the fact that CAR was susceptible to the action of trypsin, a few amounts of the latter enzyme and a precise dilution did not impair its correct detection by flow cytometry. This technique was used to demonstrate that the density of cells did not influence the expression of CAR at the cell surface. CaCo-2 cells express high levels of CAR and DAF at their surface. Flow cytometry, if used adequately, represents a helpful tool for the study of the interactions between these cells and various viral targets.

scholarly journals Cell-surface expression of PrPC and the presence of scrapie prions in the blood of goats

2012 ◽  
Vol 93 (5) ◽  
pp. 1127-1131 ◽  
Author(s):  
Rohana P. Dassanayake ◽  
David A. Schneider ◽  
Lynn M. Herrmann-Hoesing ◽  
Thomas C. Truscott ◽  
William C. Davis ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Whole Blood ◽  
Surface Expression ◽  
Infected Sheep ◽  
Cell Surface Expression ◽  
Prion Infectivity ◽  
Infected Goat ◽  
Scrapie Infection ◽  
Relative Differences

Although host-encoded prion protein (PrPC) expression in ovine PBMCs and prion infectivity in scrapie-infected sheep blood have been demonstrated, such studies have not been reported in goats. Therefore, this study characterized cell-surface expression of PrPC on PBMC subsets derived from normal goats and sheep, by flow cytometry, and determined prion infectivity in blood from a scrapie-infected goat using a transfusion bioassay in goat kids. Cell-surface PrPC expression was detected on all subsets of goat PBMCs. The highest PrPC cell-surface expression was found in CD2+ T lymphocytes in goats. Transmission of infection was detected in all three recipients who received whole blood from a goat with classical scrapie. It was concluded that caprine PBMCs express PrPC similarly to sheep but with relative differences among PBMCs subsets, and that blood-borne infectious prions can be detected in scrapie-infected goats. Thus, similar to sheep, goat blood may be a suitable diagnostic target for the detection of scrapie infection.

A Simple and Rapid Method to Determine GPIIb/IIIa-Receptor Inhibitor Activity in Whole Blood

1999 ◽  
Vol 19 (03) ◽  
pp. 134-138
Author(s):  
Gitta Kühnel ◽  
A. C. Matzdorff
Keyword(s):  
Flow Cytometry ◽  
Platelet Count ◽  
Blood Sample ◽  
Platelet Activation ◽  
Whole Blood ◽  
Receptor Occupancy ◽  
Aggregate Formation ◽  
Inhibitor Activity ◽  
Receptor Inhibitor

SummaryWe studied the effect of GPIIb/IIIa-inhibitors on platelet activation with flow cytometry in vitro. Citrated whole blood was incubated with increasing concentrations of three different GPIIb/IIIa-inhibitors (c7E3, DMP728, XJ757), then thrombin or ADP were added and after 1 min the sample was fixed. Samples without c7E3 but with 0.1 U/ml thrombin had a decrease in platelet count. Samples with increasing concentrations of c7E3 had a lesser or no decrease in platelet count. The two other inhibitors (DMP 725, XJ757) gave similar results. GPIIb/IIIa-inhibitors prevent aggregate formation and more single platelets remain in the blood sample. The agonist-induced decrease in platelet count correlates closely with the concentration of the GPIIb/IIIa inhibitor and receptor occupancy. This correlation may be used as a simple measure for inhibitor activity in whole blood.

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