Calibrated automated thrombogram
The Calibrated Automated Thrombogram is a laboratory instrument designed to measure the thrombin generation potential in a sample. It provides quantitative data on the kinetics of thrombin generation, offering insights into the hemostatic and thrombotic status of the sample.
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8 protocols using calibrated automated thrombogram
Thrombin Generation Assay with Reversal Agents
Thrombogram Analysis of Platelet-Poor Plasma
Thrombin Generation Assay in Platelet-Poor Plasma
To prepare PPP, whole blood was collected through approved IRB protocol HSC‐MS‐09‐0314 from eight healthy volunteers into 4.5 ml vacutainers containing 3.2% sodium citrate. Samples were centrifuged for 10 minutes at 200g and platelet rich plasma supernatant was centrifuged again for 10 minutes at 2,000g to obtain PPP. Samples from each donor were pooled and aliquoted. CAT was performed using a Calibrated Automated Thrombogram (Diagnostica Stago, Parsippany NJ). In a 96‐well plate, 20 μl of phospholipid reagent was added to 72 μl of PPP and either 8 μl of CCTs suspended in PBS at concentration 106 cells per milliliter or PBS vehicle. Thrombin generation was initiated with 20 μl of 0.1 M calcium chloride and fluorogenic substrate solution. Samples were performed in duplicate. Reactions were measured using a Fluoroskan Ascent plate reader (ThermoFisher, Waltham, MA).
Thrombin Generation Assay Protocol
Apolipoprotein and Coagulation Factors Profiling
All coagulation-related laboratory measurements (FVII, FVIII, FIX, FXI, vWF:Ag) were analyzed on the ACL-Top 700 Analyzer (Instrumentation Laboratory). FVIII, FIX, and FXI levels were measured using modified activated partial thromboplastin time assays with immunodepleted plasmas. Similarly, FVII was determined using a modified prothrombin time assay. vWF antigen was measured using an automated latex–enhanced immunoassay with the HemosIL vWF:Ag Reagent Kit [10 (link)].
The thrombin generation potential was assessed by the Calibrated Automated Thrombogram (Diagnostica Stago) according to the specifications of the manufacturer [20 (link)]. The fluorescent signal representing generated thrombin was monitored in a Fluoroskan Ascent fluorometer (Thermo Fisher Scientific), and the parameters were calculated with the Thrombinoscope software (Thrombinoscope BV). Endogenous thrombin potential (ETP) was the parameter determined through the thrombin generation assay [11 (link),15 (link)].
Calibrated Automated Thrombogram Analysis
Comprehensive Blood Coagulation Analysis
Thrombin Generation Assay Protocol
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