Instrument and technique for the in vitro screening of platelet activation from whole blood samples.
The measurement of platelet activation is very difficult to accomplish clinically as platelets are readily activated by in vitro manipulations. Although techniques such as platelet aggregation and flow cytometry exist to estimate platelet function, important limitations prevent these techniques to be widely accepted. In this study, low-fouling surfaces used to limit ex vivo platelet activation were locally bioactivated to rapidly detect platelet activation from whole blood through the selective local adhesion and aggregation of artificially activated platelets. To achieve this result, a fabrication method was developed to create arrays of anti-CD62 and anti-CD61 proteins covalently immobilized on substrates covered by low-fouling graft layers. Moreover, to further limit ex vivo platelet activation and to obtain reproducible results, a custom-made flow chamber was designed and fabricated with the help of computer-assisted mathematical modeling to create defined shear environments. This diagnostic instrument has the potential to allow the rapid estimation of platelet activation levels in whole blood.
Martin
Yves
Y
Laboratoire de Bioingénierie et de Biophysique de l'Université de Sherbrooke, Department of Chemical Engineering, Université de Sherbrooke, 2500 Boulevard de l'Université, Sherbrooke, Quebec, Canada. yves.martin@usherbrooke.ca
Lépine
Mariette
M
Bannari
Abdelfettah
A
Vermette
Patrick
P
D001792:Blood Platelets / Q000166:cytology* / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D015539:Platelet Activation / Q000502:physiology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity