We use cookies to understand how you use our site and to improve your experience. This includes personalizing content and advertising. To learn more, click here. By continuing to use our site, you accept our use of cookies. Cookie Policy.

Features Partner Sites Information LinkXpress
Sign In
Advertise with Us
PURITAN MEDICAL

Download Mobile App




Flow Cytometry Technique Assessed for Signaling Pathways

By LabMedica International staff writers
Posted on 24 Apr 2018
Print article
Image: The Navios clinical flow cytometer (Photo courtesy of Beckman Coulter).
Image: The Navios clinical flow cytometer (Photo courtesy of Beckman Coulter).
Platelet aggregation is a key parameter in arterial thrombosis, leading to myocardial infarction or ischemic stroke. Platelets have several receptors sensitive to different stimuli and drugs that modulate aggregation with potential critical clinico-pathological significance.

Platelet intracellular signaling is essential for platelet function but difficult to study because of the small size and the need for delicate manipulation of platelets and because the reference techniques require large cell volumes and sample preparation. Flow cytometry is often used in routine clinical laboratories.

A team of scientists collaborating with CHU Saint-Etienne, Hôpital Nord (Saint-Etienne, France) assessed degree of platelet vasodilator-stimulated phosphoprotein (VASP) phosphorylation on patients hospitalized for a non-cardio-embolic ischemic stroke, included in an observational study. The patients, 12 men, seven women with a mean age of 72 years, were enrolled in the study between November 2014 and September 2015, and were treated orally with a regular daily dose of 75 mg of clopidogrel for at least five days to be at steady state, before blood was drawn.

The VASP assay was performed using the PLT VASP/P2Y12 kit. In addition, the Platelet PerFix flow cytometry method was developed from the PerFix EXPOSE kit provided for leukocytes. The team adapted the manufacturer's instruction for this kit. The fixation, permeabilization, staining and washing reagents of the kit were used. Each step was performed at room temperature. Platelets were analyzed on a Navios cytometer using Navios software (Beckman-Coulter) within one hour after the preparation was completed. Maximum platelet aggregation induced by adenosine diphosphate (ADP) was measured by Light Transmittance Aggregometry or LTA.

The investigators found poor agreement between prevention by adenosine diphosphate (ADP) of PGE1-induced cAMP-mediated VASP phosphorylation and ADP induced aggregation assessed by Light Transmittance Aggregometry. They did find a significant correlation between the PLT VASP/P2Y12 kit and the PerFix EXPOSE kit. The PerFix EXPOSE kit may also be helpful to monitor adverse effects of second-generation tyrosine kinase inhibitors on platelets as this new method can detect low responders to clopidogrel. The study was published originally on-line on February 16, 2018, in the journal Practical Laboratory Medicine.

Related Links:
CHU Saint-Etienne

Platinum Member
COVID-19 Rapid Test
OSOM COVID-19 Antigen Rapid Test
Specimen Collection & Transport
Anti-Cyclic Citrullinated Peptide Test
GPP-100 Anti-CCP Kit
Gold Member
Real-time PCR System
GentierX3 Series

Print article

Channels

Clinical Chemistry

view channel
Image: Reaching speeds up to 6,000 RPM, this centrifuge forms the basis for a new type of inexpensive, POC biomedical test (Photo courtesy of Duke University)

POC Biomedical Test Spins Water Droplet Using Sound Waves for Cancer Detection

Exosomes, tiny cellular bioparticles carrying a specific set of proteins, lipids, and genetic materials, play a crucial role in cell communication and hold promise for non-invasive diagnostics.... Read more

Molecular Diagnostics

view channel
Image: The study showed the blood-based cancer screening test detects 83% of people with colorectal cancer with specificity of 90% (Photo courtesy of Guardant Health)

Blood Test Shows 83% Accuracy for Detecting Colorectal Cancer

Colorectal cancer is the second biggest cause of cancer deaths among adults in the U.S., with forecasts suggesting 53,010 people might die from it in 2024. While fewer older adults are dying from this... Read more

Microbiology

view channel
Image: The new platform is designed to perform blood-based diagnoses of nontuberculosis mycobacteria (Photo courtesy of 123RF)

New Blood Test Cuts Diagnosis Time for Nontuberculous Mycobacteria Infections from Months to Hours

Breathing in nontuberculous mycobacteria (NTM) is a common experience for many people. These bacteria are present in water systems, soil, and dust all over the world and usually don't cause any problems.... Read more

Industry

view channel
Image: These new assays are being developed for use on the recently introduced DxI 9000 Immunoassay Analyzer (Photo courtesy of Beckman Coulter)

Beckman Coulter and Fujirebio Expand Partnership on Neurodegenerative Disease Diagnostics

Beckman Coulter Diagnostics (Brea, CA, USA) and Fujirebio Diagnostics (Tokyo, Japan) have expanded their partnership focused on the development, manufacturing and clinical adoption of neurodegenerative... Read more
Copyright © 2000-2024 Globetech Media. All rights reserved.