A new, simplified technique for the radioautography of freeze-fracture specimens has been developed using freeze-fractured liver tissues from rats given injection of ('3)H-thymidine following partial hepatectomy, or intestine from rats given injection of ('3)H-fucose. / The specimens are fractured, replicated with platinum and carbon, and freeze-dried in-vacuo in about six hours. The replicated side of the specimen is covered with Ilford L(,4) emulsion for radioautography. After exposure and photographic processing, the emulsion is protected in three ways before removing the tissue underlying the replica by digestion in sodium hypochlorite solution: its silver grains are coated with gold, a carbon layer is evaporated over it, and grease is applied over the carbon layer. The specimen is then taken through hypochlorite and the grease dissolved in Freon 14 before it is mounted on grid for electron microscopic examination. / The validity of the technique was tested in two ways. First, freeze-fracture radioautography of the liver tissues from rats given injections of ('3)H-thymidine showed a label distribution identical to that in routine radioautographs. Secondly, the localization of ('3)H-fucose was traced to the E-face of intestinal microvilli. These preliminary results demonstrate the potential of the described technique.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.68570 |
| Date | January 1981 |
| Creators | Kan, Frederick Wai Kwong |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Anatomy) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 000112463, proquestno: AAINK51994, Theses scanned by UMI/ProQuest. |
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