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Platelet meeting Feb 24

Platelet meeting Feb 24. Sample preparation. Platelets suspension from citrated blood Centrifuged 2x at 800g/1.5 min to collect PRP PRP is centrifuged at 680g for 15 min, supernatant removed and resuspended in citrated Tyrode’s x 2 Last resuspension in Tyrode’s with Apyrase

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Platelet meeting Feb 24

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  1. Platelet meeting Feb 24

  2. Sample preparation • Platelets suspension from citrated blood • Centrifuged 2x at 800g/1.5 min to collect PRP • PRP is centrifuged at 680g for 15 min, supernatant removed and resuspended in citrated Tyrode’s x 2 • Last resuspension in Tyrode’s with Apyrase • Two microcentrifuge tubes are prepared • Platelet suspension + ACD • Platelets suspension + 12 uM TRAP • After a couple of minutes ACD is added as well to balance the conductivity of the samples

  3. Measurement setup • All syringes, tubings, fixture and chip is treated with 5% BSA in PBS for at least 1h • Two plastic syringes are connected to a 3-way valve that connects to the fixture/chip • One syringe per sample • Oil as sheath flow • Resulting in pulsations which change the core width and thus widens the amplitude distribution

  4. Detection software • Now only detects peaks in the lowest frequency • Uses the detected peak coordinates to extract data from other frequencies • Phase, opacity and magnitude is calculated from the X and Y data

  5. Peak widening due to pulsations Two kinds of particles in PBS with oil or water as sheath

  6. Peak widening due to pulsations Two kinds of particles in PBS with oil or water as sheath

  7. Data trace – particles and platelets

  8. Phase vs Magnitude

  9. Opacity vs Magnitude

  10. PCA analysis in JMP Platelets and 10 um polystyrene particles

  11. PCA analysis in JMP Platelets only, 10 um polystyrene particles excluded

  12. Whole blood

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