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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Venter, Chantelle Van Der Merwe, Christiaan Frederick Oberholzer, Hester Magdalena Bester, Megan Jean Taute, Helena |
| Description | Country affiliation: South Africa Author Affiliation: Venter C ( Department of Anatomy, Faculty of Health Sciences, University of Pretoria, South Africa.) |
| Abstract | Fixation of biological samples is an important process especially related to histological and ultrastructural studies. Chemical fixation was the primary method of fixing tissue for transmission electron microscopy for many years, as it provides adequate preservation of the morphology of cells and organelles. High pressure freezing (HPF) and freeze substitution (FS) is a newer alternative method that rapidly freezes non-cryoprotected samples that are then slowly heated in the FS medium, allowing penetration of the tissue to insure adequate fixation. This study addresses several issues related to tissue preservation for electron microscopy. Using mice liver tissue as model the difference between samples fixed chemically or with HPF immediately after excision, or stored before chemical or HPF fixation were tested with specific focus on the nuclear membrane. Findings are that immediate HPF is the method of choice compared to chemical fixation. Of the chemical fixatives, immediate fixation with 2.5% glutaraldehyde (GA)/formaldehyde (FA) is the best in preserving membrane morphology, 2.5% GA can be used as alternative for stored and then chemically processed samples, with 10% formalin being suitable as a storage medium only if followed by HPF fixation. Overall, storage leads to lower ultrastructural preservation, but HPF with FS can minimize these artifacts relative to other processing protocols. |
| File Format | HTM / HTML |
| ISSN | 1059910X |
| Issue Number | 9 |
| Volume Number | 76 |
| e-ISSN | 10970029 |
| Journal | Microscopy Research and Technique |
| Language | English |
| Publisher | Wiley-Liss |
| Publisher Date | 2013-09-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Chemistry Techniques Freeze Substitution Methods Freezing Hydrostatic Pressure Tissue Fixation Animals Fixatives Metabolism Liver Ultrastructure Mice Microscopy, Electron, Transmission Nuclear Envelope Comparative Study Evaluation Studies Journal Article |
| Content Type | Text |
| Resource Type | Article |
| Subject | Anatomy Histology Instrumentation Medical Laboratory Technology |
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