The Effects of Shear Stress on Platelets Treated with Antibodies Against Mechanosensory Receptors

Abstract

Source: Quach, M. E. et al., A Uniform Shear Assay for Human Platelet and Cell Surface Receptors via Cone-plate Viscometry. J. Vis. Exp. (2019)

This video demonstrates the method to assess the impact of shear stress on platelets. Antibody treatment induces dissociation and conformational changes in mechanoreceptor complexes, activating the platelets and expressing markers on the surface. These markers are observed via immunostaining and flow cytometry.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Blood Draw and Platelet Isolation Draw human blood from consenting healthy adult donors via venipuncture on the day of the experiment into 3.8% trisodium citrate. One 4.5 mL tube of blood is sufficient to yield enough platelet-rich plasma (PRP) for 20-25 conditions in donors whose platelet counts are close to 250 x 103 per μL…

Materials

APC anti-human CD62P (P-Selectin)	BioLegend	304910
Brookfield Cap 2000+ Viscometer	Brookfield	–
FITC-conjugated Erythrina cristagalli lectin (ECL)	Vector Labs	FL-1141
Pooled Normal Human Plasma	Precision Biologic	CCN-10
Vacutainer Light Blue Blood Collection Tube (Sodium Citrate)	BD	369714
Vacutainer Blood Collection Set, 21G x ¾" Needle	BD	367287