Experimental and imaging techniques for examining fibrin clot structures in normal and diseased states

J Vis Exp. 2015 Apr 1:(98):e52019. doi: 10.3791/52019.

Abstract

Fibrin is an extracellular matrix protein that is responsible for maintaining the structural integrity of blood clots. Much research has been done on fibrin in the past years to include the investigation of synthesis, structure-function, and lysis of clots. However, there is still much unknown about the morphological and structural features of clots that ensue from patients with disease. In this research study, experimental techniques are presented that allow for the examination of morphological differences of abnormal clot structures due to diseased states such as diabetes and sickle cell anemia. Our study focuses on the preparation and evaluation of fibrin clots in order to assess morphological differences using various experimental assays and confocal microscopy. In addition, a method is also described that allows for continuous, real-time calculation of lysis rates in fibrin clots. The techniques described herein are important for researchers and clinicians seeking to elucidate comorbid thrombotic pathologies such as myocardial infarctions, ischemic heart disease, and strokes in patients with diabetes or sickle cell disease.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Anemia, Sickle Cell / blood
  • Blood Chemical Analysis / methods*
  • Factor XIIIa / chemistry
  • Factor XIIIa / metabolism
  • Fibrin / analysis*
  • Fibrin / chemistry
  • Fibrinogen / chemistry
  • Fibrinogen / metabolism
  • Fibrinolysis
  • Humans
  • Microscopy, Confocal / methods
  • Myocardial Infarction / blood
  • Myocardial Ischemia / blood
  • Stroke / blood
  • Thrombin / chemistry
  • Thrombin / metabolism
  • Thrombosis / blood

Substances

  • Fibrin
  • Fibrinogen
  • Factor XIIIa
  • Thrombin