Comparing different preparation methods to study human fibrin fibers and platelets using TEM
Article first published online: 29 DEC 2011
Copyright © 2011 Wiley Periodicals, Inc.
Microscopy Research and Technique
Volume 75, Issue 6, pages 801–806, June 2012
How to Cite
Buys, A. V. and Pretorius, E. (2012), Comparing different preparation methods to study human fibrin fibers and platelets using TEM. Microsc. Res. Tech., 75: 801–806. doi: 10.1002/jemt.21129
- Issue published online: 21 MAY 2012
- Article first published online: 29 DEC 2011
- Manuscript Accepted: 31 OCT 2011
- Manuscript Received: 22 SEP 2011
- Medical Research Council of South Africa
- South African National Blood Services
- fibrin fibers;
- fixation methods
For the study of cellular ultrastructure, the sample needs to be stabilized by fixation, with the ultimate aim to preserve the native tissue organization and to protect the tissue against later stages of preparation. Chemical and freezing fixation are most used, and chemical fixation employs agents that permeate tissues and cells by diffusion and covalently bind with their major biochemical constituents to fix them. Most widely used chemical fixatives are aldehydes, e.g., formaldehyde and glutaraldehyde, which are noncoagulating, crosslinking agents. Cryofixation methods for ultrastructural studies are also popular, and high-pressure freezing immobilizes all cell constituents and arrests biological activity by removing the thermal energy from the system. In the current research, we used platelet-rich plasma (PRP) to study expansive fibrin fibers and platelet ultrastructure to compare the two fixation techniques. We also used thrombin and calcium chloride as a clotting agent to determine the technique most suitable for the formation of extensive fibrin networks. Chemically fixated fibrin fibers were more compact and condensed and also showed a banding pattern on longitudinal sections. High-pressure frozen samples were more dispersed while platelets fixated showed better preserved cellular membranes and organelle structure. PRP coagulated by addition of CaCl2 showed blood platelets that are noticeably more activated compared with PRP; however, with thrombin, a sharp ultrastructure was seen. We conclude that PRP mixed with thrombin, and freeze substituted, is the most suitable method for the study of extensive fibrin fibers as well as platelets. Microsc. Res. Tech. 75:801–806, 2012. © 2011 Wiley Periodicals, Inc.