A simple method for exposing and examining the interior of fragile biological materials
| Autor: | Arthur J. Wasserman, Y. Pedro Kato, Frederick H. Silver |
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| Rok vydání: | 1990 |
| Předmět: | |
| Zdroj: | Proceedings, annual meeting, Electron Microscopy Society of America. 48:858-859 |
| ISSN: | 2690-1315 0424-8201 |
| DOI: | 10.1017/s0424820100161850 |
| Popis: | Examining the inside of delicate and fragile dry biomaterials is difficult because they are vulnerable to mechanical damage. Compression and sheering of a sample during exposure of the interior can produce artifacts making interpretation of the ultrastructure difficult. In this report a simple method for exposing and retaining the interior substructure of delicate specimens and mounting them for scanning electron microscopic (SEM) observation is described.Collagen fibers were prepared as described previously. In brief, 1% collagen dispersion, prepared from bovine hide, was extruded through polyethylene tubing with an inner diameter of 0.28 mm into a 37°C, pH 7.5, fiber formation buffer. After 45 mins in the buffer, the fibers were rinsed in isopropyl alcohol for 4 hrs followed by distilled water for 20 mins. The fibers were then crosslinked.The interior of collagen fibers were exposed by deep freezing 1 cm segments of fiber in liquid nitrogen. Using iris scissors the first and last piece of each segment (which contained ends previously exposed to the atmosphere) were snapped away and discarded. Each frozen segment was then snapped in half. By this method each half of the original 1 cm segment had a freshly cleaved top and bottom surface. The segments were transferred to an aluminum foil pouch (in liquid nitrogen). |
| Databáze: | OpenAIRE |
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