Analyzing fibrin clot structure using a microplate reader

Blood Coagul Fibrinolysis. 2002 Sep;13(6):533-9. doi: 10.1097/00001721-200209000-00008.

Abstract

Fibrin clot structure studies are often performed using optical methods. For example, the clot's fiber structure can be assessed by measuring light scattering as a function of wavelength. From these measurements, one can calculate the mass/length ratio (mu), a relative measure of fibrin thickness. Fiber thickness has important functional correlates in terms of clot stability and resistance to fibrinolysis. Typically, measurements to calculate mass/length ratios are carried out on high-end spectrophotometers. However, limitations of this instrument include the large sample volume required and the inability to read multiple samples at one time. To circumvent these limitations, a plate-reading spectrophotometer is more commonly used to monitor clot formation; increases in absorbance indicate clot formation, while decreases indicate clot lysis. However, it is unclear whether plate-reading spectrophotometers can be used to quantitatively evaluate fibrin fiber structure. In the current study, we compared spectrophotometric analysis of fibrin gels on single-sample and plate-reading spectrophotometers. Results show that a plate-reading spectrophotometer does not give accurate measurements of the fiber mass/length ratio. However, the plate-reading spectrophotometer can provide a qualitative measure of fiber structure for both purified fibrinogen and plasma. We suggest that plate-reading spectrophotometers can provide a convenient, rapid, and inexpensive means of analyzing fibrin clot structure.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Calcium / pharmacology
  • Dextrans / pharmacology
  • Dose-Response Relationship, Drug
  • Fibrin / analysis*
  • Fibrin / metabolism
  • Fibrin / ultrastructure*
  • Fibrinolysis
  • Humans
  • Kinetics
  • Reproducibility of Results
  • Spectrum Analysis / instrumentation
  • Spectrum Analysis / standards
  • Thrombin / pharmacology

Substances

  • Dextrans
  • Fibrin
  • Thrombin
  • Calcium