Pfa 100

Manufactured by Siemens

Sourced in Germany, United States

The PFA-100 is a lab equipment product manufactured by Siemens. It is designed to measure platelet function in blood samples. The device uses a specialized cartridge to analyze the ability of platelets to aggregate in response to specific agonists, providing information about the platelet's activity and function.

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38 protocols using pfa 100

Platelet Function Analysis for Hemostasis

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There are numerous recommendations to also use desmopressin acetate as a treatment for platelet function disorders or mild thrombocytopenia. Desmopressin acetate is used to improve primary haemostasis—not because of the enhanced aggregability of the platelets, but rather due to their increased adhesiveness [16 ].
Platelet function was tested using platelet function analysis (PFA-100, Dade Behring, Germany), which measures the time required for a platelet plug to form and seal a semipermeable membrane. This membrane is coated in collagen. As under physiological conditions, it serves as an initial matrix for platelet adhesion in relation to the von Willebrand factor found in the tested blood. During the test, the platelets bind directly with the collagen, which in turn leads to platelet activation. Adherent platelets are degranulated on contact with the agonists' epinephrine and adenosine phosphate, which are also found in the membrane. Activation of free-flowing platelets and blood clot aggregation is the outcome. Consequently, the opening in the membrane gradually closes (Figure 4) [17 ]. The standard values for membrane closure range from 68 to 120 seconds for adenosine phosphate and 54 to 165 seconds for epinephrine. The uneventful use of PFA-100 in neurosurgical patients is documented in the literature [18 (link)].

Kapapa T., Röhrer S., Struve S., Petscher M., König R., Wirtz C.R, & Woischneck D. (2014). Desmopressin Acetate in Intracranial Haemorrhage. Neurology Research International, 2014, 298767.

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Anticoagulation and Antiplatelet Regimen for CF-LVAD Patients

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All the CF-LVAD patients in the study were treated with our standard anticoagulation and antiplatelet regimen. Anticoagulation was initiated with a titrated heparin dose with the goal for partial thromboplastin time of 40–45s once chest drainage was less than 30 mL/h for at least 4 hours. Thereafter the goal was aimed to have an anti-Xa activity level of 0.1–0.15 U/mL. The anticoagulation medication was subsequently converted to warfarin with a targeted international normalized ratio (INR) for the HeartMate II (1.8–2.3), the Jarvik 2000 (2.0–3.0) and the HeartWare HVAD (2.0–3.0). Antiplatelet agents were added to the anticoagulation regimen and the dosage was titrated based on measurements of platelet function using a platelet function analyzer (PFA-100^®, Dade Behring, Inc, Deerfield, IL) and thrombelastogram (TEG) (TEG® 5000 Thrombelastograph® Hemostasis Analyzer System, Haemonetics Corporation, Braintree, MA). All the patients received pentoxifylline to improve red blood cell deformability in the hope of mitigating shear-induced hemolysis. Although individual patient anticoagulation/antiplatelet regimen might vary day-to-day, a standard strategy was followed for all the patients.

Mondal N.K., Chen Z., Trivedi J.R., Sorensen E.N., Pham S.M., Slaughter M.S., Griffith B.P, & Wu Z.J. (2017). Oxidative stress induced modulation of platelet Integrin α2bβ3 expression and shedding may predict the risk of major bleeding in heart failure patients supported by continuous flow left ventricular assist devices. Thrombosis research, 158, 140-148.

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Assessing Platelet Function via PFA-100

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The whole blood was immediately anticoagulated with 3.2% buffered citrate and using the PFA-100™ (Dade Behring, Marburg, Germany) to check platelet function. The blood sample was sent for investigation within 4 hours after collection. PFA-100 monitored platelet interaction on two cartridges. Both held a membrane with a 150 μm aperture and separately coated with collagen and either epinephrine or ADP for CEPI or CADP cartridge. This instrument provided a closure time which was determined by the time required for the platelet plug to occlude the aperture and was expressed in seconds. The normal range of the closure time is 85 to 165 seconds for CEPI and 71 to 118 seconds for CADP in the general population.

Chen C.Y., Liou H.H., Chang M.Y., Wang H.H., Lee Y.C., Ho L.C., Lin T.M, & Hung S.Y. (2022). The use of a low-flux hemo-dialyzer is associated with impaired platelet aggregation in patients undergoing chronic hemodialysis. Medicine, 101(43), e31623.

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Platelet Aggregation and Bleeding Time in Dogs

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Platelet aggregation time (PAT) was assessed after platelet stimulation using a mixture of collagen and adrenaline (PFA-100®, Dade-Behring, Paris) with a maximum cut-off of 300 s. In dogs, a normal PAT value is considered < 210 s [49 (link)], whereas internal validation according to our experimental conditions indicated a normal PAT value as < 213 s. Buccal mucosal bleeding time was performed in dogs in which platelet aggregation time was increased and it was considered normal if it was below 4 min.

Monteiro B.P., Lambert C., Bianchi E., Genevois J.P., Soldani G, & Troncy E. (2019). Safety and efficacy of reduced dosage ketoprofen with or without tramadol for long-term treatment of osteoarthritis in dogs: a randomized clinical trial. BMC Veterinary Research, 15, 213.

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Anticoagulation and Antiplatelet Strategies for CF-LVAD

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After CF-LVAD implantation, anticoagulation was initiated with a titrated heparin dose with the goal for partial thromboplastin time of 40-45s once chest drainage was less than 30 mL/h for at least 4 hours. Thereafter the goal was aimed to have an anti-Xa activity level of 0.1-0.15 U/mL. The anticoagulation medication was subsequently converted to warfarin with a targeted international normalized ratio (INR) from 1.8 to 2.3 for the HeartMate II, 2 to 3 for the Jarvik 2000 and the HeartWare HVAD. Antiplatelet agents were added to the anticoagulation regimen and the dosage was titrated based on measurements of platelet function using a platelet function analyzer (PFA-100^®, Dade Behring, Inc, Deerfield, IL) and thrombelastogram (TEG) (TEG® 5000 Thrombelastograph® Hemostasis Analyzer System, Haemonetics Corporation, Braintree, MA). All the patients received pentoxifylline to improve red blood cell (RBC) deformability in the hope of mitigating shear-induced hemolysis.

Mondal N.K., Sorensen E.N., Pham S.M., Koenig S.C., Griffith B.P., Slaughter M.S, & W Z.J. (2015). Systemic inflammatory response syndrome in end stage heart failure patients following continuous-flow left ventricular assist devices implantation: differences in plasma redox status and leukocyte activation. Artificial organs, 40(5), 434-443.

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Platelet Function Assessment Protocol

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To assess platelet function, we performed arachidonic acid (AA)‐induced whole‐blood aggregometry and measured platelet function under high shear rates, both standard methods to assess responsiveness to ASA treatment 14.
Whole‐blood aggregation was determined using the multiple electrode aggregometry (MEA) on the Multiplate Analyzer (Dynabyte Medical, Munich, Germany). AA‐ and ADP‐induced platelet aggregation were performed as explained previously 15 (Appendix S1). Based on other trials, we chose a cut‐off of > 30 U (arbitrary units) 16, 17, 18.
The platelet function analyzer‐100 (PFA‐100; Dade Behring, Marburg, Germany) was used for measuring platelet function under high shear rates (5000–6000 s⁻¹) as described previously 19 (Appendix S1). Collagen/epinephrine (PFA‐EPI) and collagen/ADP (PFA‐ADP) coated cartridges were used. As a cut‐off value for HTPR, a closure time < 193 s for PFA‐EPI was recommended 20.
Thromboxane B2 (TXB2) levels were measured by enzyme‐linked immunoassay (ELISA; Cayman Chemical, Ann Arbor, MI, USA) as previously reported 20.

Schoergenhofer C., Hobl E., Schwameis M., Gelbenegger G., Staudinger T., Heinz G., Speidl W.S., Zauner C., Reiter B., Lang I, & Jilma B. (2017). Acetylsalicylic acid in critically ill patients: a cross‐sectional and a randomized trial. European Journal of Clinical Investigation, 47(7), 504-512.

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Anticoagulation Regimen for CF-LVAD Patients

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After CF-LVAD implantation, anticoagulation was initiated with a titrated heparin dose with the goal for partial thromboplastin time of 40–45s once chest drainage was less than 30 mL/h for at least 4 hours. Thereafter the goal was aimed to have an anti-Xa activity level of 0.1–0.15 U/mL. The anticoagulation medication was subsequently converted to warfarin with a targeted international normalized ratio (INR) from 1.8 to 2.3 for the HeartMate II, 2 to 3 for the Jarvik and the HeartWare. Antiplatelet agents were added to the anticoagulation regimen and the dosage was titrated based on measurements of platelet function using a platelet function analyzer (PFA-100® (Dade Behring, Inc, Deerfield, IL) and thrombelastogram (TEG) (TEG® 5000 Thrombelastograph® Hemostasis Analyzer System, Haemonetics Corporation, Braintree, MA). All the patients received pentoxifylline to improve RBC deformability in the hope of mitigating shear-induced hemolysis.

Mondal N.K., Sorensen E.N., Feller E.D., Pham S.M., Griffith B.P, & Wu Z.J. (2015). Comparison of Intraplatelet Reactive Oxygen Species, Mitochondrial Damage and Platelet Apoptosis after Implantation of Three Continuous Flow Left Ventricular Assist Devices: HeartMate II, Jarvik 2000 and HeartWare. ASAIO journal (American Society for Artificial Internal Organs : 1992), 61(3), 244-252.

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Anticoagulation and Antiplatelet Therapy Post-CF-LVAD

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After CF-LVAD implantation, anticoagulation was initiated with a titrated heparin dose with the goal for partial thromboplastin time of 40-45s once chest drainage was less than 30 mL/h for at least 4 hours. Thereafter the goal was aimed to have an anti-Xa activity level of 0.1-0.15 U/mL. The anticoagulation medication was subsequently converted to warfarin with a targeted international normalized ratio (INR) from 1.8 to 2.3 for the HeartMate II, 2 to 3 for the Jarvik and the HeartWare. Antiplatelet agents were added to the anticoagulation regimen and the dosage was titrated based on measurements of platelet function using a platelet function analyzer (PFA-100^® (Dade Behring, Inc, Deerfield, IL) and thrombelastogram (TEG) (TEG^® 5000 Thrombelastograph^® Hemostasis Analyzer System, Haemonetics Corporation, Braintree, MA). All the patients received pentoxifylline to improve RBC deformability in the hope of mitigating shear-induced hemolysis.

Mondal N.K., Sorensen E.N., Feller E.D., Pham S.M., Griffith B.P, & Wu Z.J. (2015). Systemic Inflammatory Response Syndrome after Contentious-Flow Left Ventricular Assist Device Implantation and Change in Platelet Mitochondrial Membrane Potential. Journal of cardiac failure, 21(7), 564-571.

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Anticoagulation and Antiplatelet Therapy for LVAD Patients

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Anticoagulation was initiated with a titrated heparin dose with the goal for partial thromboplastin time of 40–45s once chest drainage was less than 30 mL/h for at least 4 hours. Thereafter the goal was aimed to have an anti-Xa activity level of 0.1–0.15 U/mL. The anticoagulation medication was subsequently converted to warfarin with a targeted international normalized ratio (INR) for the HeartMate II (1.8 – 2.3), the Jarvik 2000 (2.0 – 3.0) and the HeartWare HVAD (2.0 – 3.0). Antiplatelet agents were added to the anticoagulation regimen and the dosage was titrated based on measurements of platelet function using a platelet function analyzer (PFA-100^®, Dade Behring, Inc, Deerfield, IL) and thrombelastogram (TEG) (TEG® 5000 Thrombelastograph® Hemostasis Analyzer System, Haemonetics Corporation, Braintree, MA). All the patients received pentoxifylline to improve red blood cell deformability in the hope of mitigating shear-induced hemolysis.

Mondal N.K., Li T., Chen Z., Chen H.H., Sorensen E.N., Pham S.M., Sobieski M.A., Koenig S.C., Slaughter M.S., Griffith B.P, & Wu Z.J. (2017). Mechanistic Insight of Platelet Apoptosis leading to Non-surgical Bleeding among Heart Failure Patients supported by Continuous-Flow Left Ventricular Assist Devices. Molecular and cellular biochemistry, 433(1-2), 125-137.

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Platelet Aggregation and Adherence Assessment

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The effects of the compounds on platelet aggregation/activation and adherence in whole blood under conditions of high shear stress flow were evaluated using a collagen/epinephrine cartridge (Dade Behring, Marburg, Germany) and platelet function analyzer (PFA-100), as previously described [20 (link)]. Blood samples were drawn from healthy donors using 3.8% (w/v) sodium citrate. Aliquots of whole blood were treated with different concentrations of compounds and incubated for 5 min at room temperature. Blood samples were added to a collagen/epinephrine cartridge, and the time required for occlusion of the aperture was measured using the analyzer and reported as the closure time. Saline-treated blood was used as a control.

Choi J.H, & Kim S. (2022). In Vitro Antithrombotic, Hematological Toxicity, and Inhibitor Studies of Protocatechuic, Isovanillic, and p-Hydroxybenzoic Acids from Maclura tricuspidata (Carr.) Bur. Molecules, 27(11), 3496.

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