Mouse and human platelet (2×108 platelets/ml) aggregation was performed with washed platelets using a Chrono-log Model 700 Whole Blood/Optical Lumi-Aggregometer with AGGRO/LINK software (Chrono-log), stirring at 1200 rpm at 37°C. Adenosine diphosphate (ADP) induced aggregation was performed in the presence of human fibrinogen (0.5mg/ml, Sigma-Aldrich). Agonists were used at concentrations indicated: ADP (Chrono-log), Collagen (Chrono-log), thrombin (Roche), U46619 (Cayman Chemical Company).
Model 700 whole blood optical lumi aggregometer
Manufactured by Chrono-Log
Sourced in United States
The Chrono-log Model 700 Whole Blood/Optical Lumi-Aggregometer is a laboratory instrument designed to measure platelet aggregation and luminescence in whole blood samples. It is used to assess the functional activity of platelets in response to various agonists.
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7 protocols using model 700 whole blood optical lumi aggregometer
1
Platelet Aggregation Assay with Agonists
2
Murine Platelet Aggregation Assay
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Gel-filtered murine platelets were used to measure the platelet aggregation in response to thrombin. Platelets were diluted to the final concentration of 5 × 10 7 platelets/mL in a 300-μL reaction volume, which contains 295 μL of gel-filtered platelets and 5 μL agonist. Platelet aggregation was recorded using the CHRONO-LOG Model 700 Whole Blood/Optical Lumi-Aggregometer paired with AGGRO/LINK 8 program software (Chrono-log Corporation).
3
Platelet Aggregation Assay Methodology
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For in vitro aggregation washed platelets (300.000 cells μL−1) were pre treated with antioxidants/inhibitors at 37°C for 10 min in the presence of apyrase VII 10 U mL−1 and ASA 100 μM. Platelets were transferred into cuvettes and incubated with CaCl2 1 mM and MgSO4 1 mM for 1 min at 37°C under continuous stirring at 1000 rpm. Platelet aggregation was monitored for 5 min after the addition of the agonist, using the Born’s turbidimetric method in a four channel aggregometer (APACT 4004, Labitech and Chrono-log Model 700 Whole Blood/Optical Lumi-Aggregometer, Chrono-Log Corp.). The rate of platelet aggregation was calculated as change in percentage of transmitted light (%T), according to Born and Cross (1963) (link). For the ex vivo experiments, aggregation tests were performed using PRP in the presence/absence of apyrase VII 10 U mL−1 with platelet count adjusted at 300.000 platelets μL−1.
4
Simultaneous Measurement of ATP-Release and Platelet Aggregation
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For simultaneous measurement of the adenosine triphosphate-release and aggregation WT and reeler platelets, the CHRONO-LOG Model 700 Whole Blood/Optical Lumi Aggregometer (CHRONO-LOG Corp) was used. A total of 1×10 5 platelets/μL in presence of CaCl 2 (2 mmol/L) and fibrinogen (70 μg/mL) were measured for each sample. The warmed (37°C) platelet suspension was put into CHRONO-LOG cuvettes (312), stirred at 1000 rpm, and incubated with luciferase (0.2 μmol/L) for 2 minutes. Then, the platelets were treated with indicated agonists and inhibitors. The duration of the aggregation measurement was fixed at 10 minutes.
5
Platelet Aggregation Evaluation by Optical Lumiaggregometry
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Platelet aggregometry was performed using whole-blood optical lumiaggregometry (Model 700 Whole Blood/Optical Lumi-aggregometer, CHRONO-LOG). Maximal platelet aggregation using electrical impedance in whole-blood samples was determined after stimulation by various platelet agonists under conditions of continuous stirring (1200 rpm).19
6
Evaluating Antiplatelet Therapy Response
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Impedance aggregometry was performed to determine whether rabbits appropriately responded to dual antiplatelet therapy. Approximately 5 ml of whole blood collected from the butterfly catheter was transferred into 3.2% sodium citrate collection tubes (BD Vacutainer) and inverted several times to deter clotting. Samples were run within 3 hours of collection as per the manufacturer's guidelines. To test for platelet response to ASA and clopidogrel, 500 µl of blood was diluted with 500 µl of 0.9% saline in a disposable cuvette with a siliconized stir bar. Collagen (1-5 µg/ml) and adenosine diphosphate (5-10 µM) were subsequently added to the sample of whole blood to test for responsiveness to ASA and clopidogrel, respectively. Samples were then analyzed using a Model 700 Whole Blood/Optical Lumi-Aggregometer (Chrono-Log Corp.). The aggregometer was set to 37°C, a gain of 0.01 times, and a speed of 1200 RPM.
7
Platelet Aggregation Assay with Vaa-SP-VX
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Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared as described in Leonardi et al. [42 (link)]. To 250 μL of PRP, 25 μL of VaaSP-VX solution was added, resulting in a final concentration of 1 µM VaaSP-VX; it was then pre-incubated for 5 min at 37 °C. In the control experiment, 25 μL of water was added to 250 μL of PRP. Twenty-five microliters of collagen solution (DiaMed, Cressier,, Switzerland), giving a final concentration of 10 μg/mL; ristocetin (Chrono-Log Corporation, Havertown, PA, USA), resulting in a final concentration of 1 mg/mL; or ADP (Chrono-Log Corporation, USA) at a final concentration of 10 μM, were lastly added and the change in the optical density was monitored for 5 min using a Model 700 Whole Blood/Optical Lumi-Aggregometer (Chrono-Log Corporation, Havertown, PA, USA). The cuvette containing PRP corresponded to 0% light transmission and the cuvette with PPP to 100% light transmission. Blood was donated by healthy human volunteers according to permission no. 53/08/11 of the National Medical Ethics Committee of the Republic of Slovenia.
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