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Thrombodynamics® analysis takes place in a single-use reaction cuvette upon initiation with an activation insert.  A key innovation of Thrombodynamics® is the use of time-lapse video-microscopy to analyze and quantify spatiotemporal parameters of fibrin clot growth/lysis.

Thrombodynamics-Principle-of-Thrombodynamics Measurement

Thrombodynamics® is both a qualitative and quantitative evaluation of blood plasma coagulation status by means of measuring and analyzing the spatiotemporal dynamics of clot growth and lysis in a heterogeneous in vitro system.

The Thrombodynamics® method aims to imitate in vitro physiological and pathophysiological processes that occur in vivo during hemostatic plug formation or thrombosis.  The coagulation process in Thrombodynamics® starts at a localized surface which has immobilized tissue factor mimicking a damaged blood vessel wall.

Unlike other coagulation assays the fibrin clot growth process in the Thrombodynamics® assay develops in space and time rather than only in time.  The fibrin clot starts to form, growing from the tissue factor bearing surface, then propagates into the bulk of the plasma sample without interaction with the tissue factor bearing surface.

 

Competitive advantages of Thrombodynamics® test:

 

  • Ability to reveal plasma with either a high thrombotic or a bleeding risk
  • Allows for study and analysis of the efficacy of anticoagulant drugs, including the newest generation of DOACs
  • Allows for study and analysis of the efficacy of factor replacement drugs.
  • An easy to understand and interpret global hemostasis test.
  • Based on the latest findings of biological science and nanotechnology

 

  • Small 120 µL sample volume
  • 3 simple pipetting steps from plasma
  • Global assay sensitive to both hypo and hypercoaguable blood
  • 2 channels for parallel testing
  • Bar-Coded reagents
  • Specific and sensitive reagent kits
  • Windows-based software
  • Open system for research applications
  • Define new tests in the system software
  • On board thermostat for precise temperature control
  • 1x Thrombodynamics® T2F Analyzer with Power Cable
  • 1x Thrombodynamics® Laptop
  • 1x Thrombodynamics® Mouse
  • 1x USB (A-B) Interface Cable
  • 1x RJ-45 Ethernet Interface Cable
  • 1x HUB USB-LAN
  • 1x Thrombodynamics® User Manual
  • 1x Thrombodynamics® Quick-Start Guide
  • 1x Thrombodynamics® Service Kit

The Thrombodynamics® research analyzer records and measures spatiotemporal characteristics of the fibrin clot formation process in a thin layer of unstirred blood plasma.  The coagulation cascade is initiated as soon as the plasma comes into contact with the localized and immobilized tissue factor (TF) and the fibrin clot starts growing from the end face of the activation insert growing into the bulk of the plasma, mimicking damaged vasculature.  The process of fibrin clot formation is recorded by the Thrombodynamics® T2F Analyzer using an adjustable time time-lapse video microscopy by means of a dark-field light scattering method.  The obtained series of photos show how the form, size, and density of the fibrin clot changes over time.  On the basis of the recorded photos, the Thrombodynamics® Analytical Software calculates the numerical parameters of spatiotemporal dynamics of fibrin clot formation.

  • Tlag [min]: Lag Time
  • Tsp [min]: Spontaneous Clot Formation Time
  • V [µm/min]: Rate of Clot Growth
  • Vi [µm/min]: Initial Rate of Clot Growth
  • Vst [µm/min]: Stationary Rate of Clot Growth
  • CS [µm]: Clot Size
  • D [a.u.]: Clot Density

 

Parameter Definition Description Influencing Factor
Tlag [min]: Lag Time The time from the beginning of the measurement until the first significant levels of fibrin can be detected. The initiation phase of coagulation.  Analogous to the prothrombin (clotting) time. This parameter is sensitive to the initial stage of blood coagulation and reactions of the extrinsic pathway.
Tsp [min]: Spontaneous Clot Formation Time The time that spontaneous clots appear in the sample which has had no contact with the activator insert.  Defined as the time to spontaneous clots reaches 5% of the overall measurement region. Characterizes clotting that is independent of the activator surface.  0 under normal conditions. Spontaneous clotting is induced by circulating activators, active coagulation factors, and microparticles.
V [µm/min]: Rate of Clot Growth The average rate of clot growth.  The default range is during the interval of 15-25 minutes after Tlag but is automatically adjusted in the presence of spontaneous clots. Characterizes the propagation stage of blood coagulation This parameter is sensitive to all coagulation cascade reactions, including the contact pathway and excluding the initiation reactions of the extrinsic pathway.
Vi [µm/min]: Initial Rate of Clot Growth The average rate of clot growth calculated on the interval 2-6 minutes following Tlag. Like Tlag, it describes the initial stages of clot growth but in terms of spatial elongation rather than local increase in thrombin concentration. This parameter is more sensitive to the reactions and components of the propagation stage than is Tlag, yet retains sensitivity to the initiation reactions.
Vst [µm/min]: Stationary Rate of Clot Growth The average rate of clot growth in the interval of 15-25 minutes after Tlag.  It is not calculated in the presence of spontaneous clots. Characterizes the propagation stage of blood coagulation This parameter is sensitive to all coagulation cascade reactions, including the contact pathway and excluding the initiation reactions of the extrinsic pathway.
CS [µm]: Clot Size The clot size at the 30th minute (default value) of measurement. An integral parameter characterizing overall fibrin clot formation reflecting overall coagulation cascade performance. Sensitive to all major components and processes of blood coagulation.
D [a.u.]: Clot Density An optical parameter equal to the intensity of light scattering from a fibrin clot and is proportional to the density of the fibrin clot mesh. Characterizes firmness and structure of the formed clot but cannot replace a direct measurement of fibrinogen. Sensitive to fibrinogen and Factor XIII activity.

 

In addition to those, a new feature of the Thrombodynamics® assay is the ability to visualize and quantify the fibrinolysis process in the presence of plasmin activators.  For this, several extra parameters can be calculated by the software:

  • LOT [min]: Lysis Onset Time
  • LP [%/min]: Lysis Progression
  • CLT [min]: Clot Lysis Time
  • LTE [min]: Lysis Time Estimation
  • LI [%]: Lysis Index

 

Parameter Definition Description
LOT [min]: Lysis Onset Time The time at which the lysis is calculated to have initiated. Describes the initial stages of fibrinolysis
LP [%/min]: Lysis Progression The linear rate of light scattering intensity decrease as a percentage of the initial value in the 5 minutes following LOT Characterizes the maximal functional activity of plasmin.
CLT [min]: Clot Lysis Time The time span from the maximum Clot density (lmax) and when it reaches 5% of lmax. Description of overall lysis activity.
LTE [min]: Lysis Time Estimation The time span between the lmax and the point when the clot density is projected to be 0. Like CLT, but extrapolated to total clot dissolution based on the calculated LP.
LI [%] The relationship of clot density at time moment LOT+5 to the lmax Characterizes the percent of clot density remaining 5 minutes after LOT

 

Dashkevich, N. M., Ovanesov, M. V., Baladina, A. N., Karamzin, S. S., Shestakov, P. I., Soshitova, N. P., . . . Ataullakhanov, F. I. (2012, November 21). Thrombin Activity Propagates in Space During Blood Coagulation as an Excitation Wave. Biophysical Journal, 103(10), 2233-2240. doi:10.1016/j.bpj.2012.10.011

Kuprash, A. D., Shibeko, A. M., Vijay, R., Nair, S. C., Srivastava, A., Ataullakhanov, F. I., . . . Balandina, A. N. (2018, December 18). Sensitivity and Robustness of Spatially Dependent Thrombin Generation and Fibrin Clot Propagation. Biophysical Journal, 115(12), 2461-2473. doi:10.1016/j.bpj.2018.11.009

Panteleev, M. A., Dashkevich, N. M., & Ataullakhanov, F. I. (2015, October). Hemostasis and thrombosis beyond biochemistry: roles of geometry, flow and diffusion. Thrombosis Research, 136(4), 699-711. doi: https://doi.org/10.1016/j.thromres.2015.07.025

Tuktamyshov, R., & Zhdanov, R. (2015). The method of in vivo evaluation of hemostais: Spatial thrombodynamics. Hematology, 20(10), 584-586. doi:10.1179/1607845415Y.0000000022

Zhalyalov, A. S., Panteleev, M. A., Gracheva, M. A., Ataullakhanov, F. I., & Shibeko, A. M. (2017). Co-ordinated spatial propagation of blood plasma clotting and fibrinolytic fronts. PLoS One, 12(7), e0180668. doi:10.1371/journal.pone.0180668