A Turbidimetric Analysis to Monitor Collagen-Induced Aggregation of Pretreated Human Platelets

In This Article

Abstract

Source: Molica, F. et al., Turbidimetry on Human Washed Platelets: The Effect of the Pannexin1-inhibitor Brilliant Blue FCF on Collagen-induced Aggregation. J. Vis. Exp. (2017)

This video demonstrates turbidimetric analysis to assess collagen-induced platelet aggregation in pre-treated, washed human platelets. Control wells show collagen-induced aggregation, whereas a high concentration of inhibitors blocks Pannexin-1 channels, thereby inhibiting collagen-induced platelet aggregation, evident from unchanged turbidity.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board.

1. Buffer Preparation for Human Blood Collection and Washed Platelet Isolation

Prepare a 100 mL aqueous solution of acid-citrate-dextrose (ACD) by dissolving 1.4 g citric acid monohydrate (C₆H₈O₇•H₂O, 66.6 mM), 2.5 g trisodium citrate dihydrate (Na₃C₆H₅O₇•2 H₂O, 85 mM) and 2 g of anhy....

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Results

Chromatography setup showing liquid phase separation, experimental workstation, detection units.
Figure 1: Aggregometry. (A) Representative image of glass cuvettes (containing a stirring magnet) used for aggregometry. The cuvette on the left shows resting platelets while the cuvette on the right illustrates platelet aggregates after co.......