Original ArticlesAnalyzing fibrin clot structure using a microplate readerWolberg, A. S.; Gabriel, D. A.; Hoffman, M.Author Information A. S. Wolberg and M. Hoffman are with the Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA; D. A. Gabriel and M. Hoffman are with the Department of Medicine and Center for Thrombosis and Hemostasis, University of North Carolina, Chapel Hill, North Carolina, USA; M. Hoffman is also with the Durham Veterans Affairs Medical Center, Durham, North Carolina, USA; and A. S. Wolberg is presently with the Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, North Carolina, USA. (Received 1 February 2002; revised 5 April 2002; accepted 11 April 2002) Sponsorship: This study was supported in part by grants HL09978 and HL48320 from the National Institutes of Health and 0160418U from the American Heart Association, by the Institute for Medical Research of the Durham VA Medical Center, and by the Department of Veterans Affairs. Address correspondence to Dr Maureane Hoffman, Durham VA Medical Center, 508 Fulton Street (113), Durham, NC 27705, USA. Tel: (+1) 919 286 6925; fax: (+1) 919 286 6818; e-mail: email@example.com Blood Coagulation & Fibrinolysis: September 2002 - Volume 13 - Issue 6 - p 533-539 Buy 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 (μ), 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. © 2002 Lippincott Williams & Wilkins, Inc.