Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1995 Apr;68(4):1551–1560. doi: 10.1016/S0006-3495(95)80327-9

Three-dimensional reconstruction of fibrin clot networks from stereoscopic intermediate voltage electron microscope images and analysis of branching.

T C Baradet 1, J C Haselgrove 1, J W Weisel 1
PMCID: PMC1282049  PMID: 7787040

Abstract

Fibrin polymerizes to produce branching fibers forming a three-dimensional network, which has been difficult to visualize by conventional microscopy. Three-dimensional images of whole clots at high resolution were obtained from stereo-pair intermediate-voltage electron micrographs. Computer software was developed to produce three-dimensional reconstructions of the networks in the form of a pattern of links that connect branching junctions. Network parameters were measured and analyzed to characterize the clots quantitatively. Models in which all links were moved to the origin, while preserving their orientation, allowed visualization of some network parameters and facilitated comparison of networks. Fibrin clots formed in three different conditions were analyzed and compared by these methods. Clots formed in 0.20 M saline buffer consist of fibers of uniform size, and most of the branching junctions consist of three links. Fibrin clots formed in 0.05 M saline buffer are made up of very large diameter fiber bundles with far fewer branching junctions and correspondingly longer links. Clots formed in 0.40 M saline buffer consist of very fine fibers with numerous branching junctions and very short links. In summary, the extent of lateral aggregation is directly related to the distance between branching junctions and inversely related to the total number of branching junctions. These observations must be considered in defining possible mechanisms of fibrin branching.

Full text

PDF
1551

Images in this article

1553FIGURE 1
on p.1553
1553FIGURE 2
on p.1553

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blombäck B., Carlsson K., Hessel B., Liljeborg A., Procyk R., Aslund N. Native fibrin gel networks observed by 3D microscopy, permeation and turbidity. Biochim Biophys Acta. 1989 Jul 27;997(1-2):96–110. doi: 10.1016/0167-4838(89)90140-4. [DOI] [PubMed] [Google Scholar]
  2. Blombäck B., Okada M. Fibrin gel structure and clotting time. Thromb Res. 1982 Jan 1;25(1-2):51–70. doi: 10.1016/0049-3848(82)90214-6. [DOI] [PubMed] [Google Scholar]
  3. Carr M. E., Jr, Gabriel D. A., McDonagh J. Influence of Ca2+ on the structure of reptilase-derived and thrombin-derived fibrin gels. Biochem J. 1986 Nov 1;239(3):513–516. doi: 10.1042/bj2390513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dang C. V., Shin C. K., Bell W. R., Nagaswami C., Weisel J. W. Fibrinogen sialic acid residues are low affinity calcium-binding sites that influence fibrin assembly. J Biol Chem. 1989 Sep 5;264(25):15104–15108. [PubMed] [Google Scholar]
  5. Erickson H. P., Fowler W. E. Electron microscopy of fibrinogen, its plasmic fragments and small polymers. Ann N Y Acad Sci. 1983 Jun 27;408:146–163. doi: 10.1111/j.1749-6632.1983.tb23242.x. [DOI] [PubMed] [Google Scholar]
  6. Fowler W. E., Hantgan R. R., Hermans J., Erickson H. P. Structure of the fibrin protofibril. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4872–4876. doi: 10.1073/pnas.78.8.4872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hantgan R. R., Hermans J. Assembly of fibrin. A light scattering study. J Biol Chem. 1979 Nov 25;254(22):11272–11281. [PubMed] [Google Scholar]
  8. Hantgan R., Fowler W., Erickson H., Hermans J. Fibrin assembly: a comparison of electron microscopic and light scattering results. Thromb Haemost. 1980 Dec 19;44(3):119–124. [PubMed] [Google Scholar]
  9. Hardy J. J., Carrell N. A., McDonagh J. Calcium ion functions in fibrinogen conversion to fibrin. Ann N Y Acad Sci. 1983 Jun 27;408:279–287. doi: 10.1111/j.1749-6632.1983.tb23251.x. [DOI] [PubMed] [Google Scholar]
  10. LATALLO Z. S., FLETCHER A. P., ALKJAERSIG N., SHERRY S. Inhibition of fibrin polymerization by fibrinogen proteolysis products. Am J Physiol. 1962 Apr;202:681–686. doi: 10.1152/ajplegacy.1962.202.4.681. [DOI] [PubMed] [Google Scholar]
  11. Langer B. G., Weisel J. W., Dinauer P. A., Nagaswami C., Bell W. R. Deglycosylation of fibrinogen accelerates polymerization and increases lateral aggregation of fibrin fibers. J Biol Chem. 1988 Oct 15;263(29):15056–15063. [PubMed] [Google Scholar]
  12. Medved' L., Ugarova T., Veklich Y., Lukinova N., Weisel J. Electron microscope investigation of the early stages of fibrin assembly. Twisted protofibrils and fibers. J Mol Biol. 1990 Dec 5;216(3):503–509. doi: 10.1016/0022-2836(90)90376-W. [DOI] [PubMed] [Google Scholar]
  13. Mosesson M. W., DiOrio J. P., Müller M. F., Shainoff J. R., Siebenlist K. R., Amrani D. L., Homandberg G. A., Soria J., Soria C., Samama M. Studies on the ultrastructure of fibrin lacking fibrinopeptide B (beta-fibrin). Blood. 1987 Apr;69(4):1073–1081. [PubMed] [Google Scholar]
  14. Mosesson M. W., DiOrio J. P., Siebenlist K. R., Wall J. S., Hainfeld J. F. Evidence for a second type of fibril branch point in fibrin polymer networks, the trimolecular junction. Blood. 1993 Sep 1;82(5):1517–1521. [PubMed] [Google Scholar]
  15. Mosesson M. W., Siebenlist K. R., Amrani D. L., DiOrio J. P. Identification of covalently linked trimeric and tetrameric D domains in crosslinked fibrin. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1113–1117. doi: 10.1073/pnas.86.4.1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Müller M. F., Ris H., Ferry J. D. Electron microscopy of fine fibrin clots and fine and coarse fibrin films. Observations of fibers in cross-section and in deformed states. J Mol Biol. 1984 Apr 5;174(2):369–384. doi: 10.1016/0022-2836(84)90343-7. [DOI] [PubMed] [Google Scholar]
  17. Ris H. The cytoplasmic filament system in critical point-dried whole mounts and plastic-embedded sections. J Cell Biol. 1985 May;100(5):1474–1487. doi: 10.1083/jcb.100.5.1474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Weisel J. W. Fibrin assembly. Lateral aggregation and the role of the two pairs of fibrinopeptides. Biophys J. 1986 Dec;50(6):1079–1093. doi: 10.1016/S0006-3495(86)83552-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Weisel J. W., Nagaswami C. Computer modeling of fibrin polymerization kinetics correlated with electron microscope and turbidity observations: clot structure and assembly are kinetically controlled. Biophys J. 1992 Jul;63(1):111–128. doi: 10.1016/S0006-3495(92)81594-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Weisel J. W., Nagaswami C., Makowski L. Twisting of fibrin fibers limits their radial growth. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8991–8995. doi: 10.1073/pnas.84.24.8991. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES