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On-block UV light polymerization of LR Gold-embedded flat-mounted tissue using low temperature modules
Johannes Kacza1, Andrei Nestler, Wolfgang Schilbach
1Department of Anatomy, Histology and Embryology, University of Leipzig, Germany. kacza@vetmed.uni-leipzig.de
Summary
Low temperature embedding for electron microscopy is challenging. A new UV light low temperature module (LTM) enables rapid, flat embedding of tissue specimens, improving fine structure preservation and sectioning parallel to the surface.
Area of Science:
- Materials Science
- Microscopy Techniques
- Neuroscience
Background:
- Low temperature embedding for microscopy requires oxygen-free polymerization, posing challenges for achieving flat specimens, especially for thin sections or retinal flat-mounts.
- Complete filling and tight sealing of containers are crucial but difficult under these conditions.
Purpose of the Study:
- To design and test a novel low temperature module (LTM) for UV light polymerization.
- To overcome difficulties in obtaining flat-embedded specimens for electron microscopy at low temperatures.
Main Methods:
- Development of a low temperature module (LTM) for UV light polymerization.
- Embedding vibratome sections of rat superior colliculus in LR Gold resin.
- UV curing of specimens within the LTM.
Main Results:
- The LTM facilitated instant creation of ready specimens with an almost planar surface.
- Electron microscopy confirmed good preservation of fine structure in embedded rat superior colliculus sections.
- The UV light polymerization technique proved suitable for obtaining ultrathin sections parallel to the embedded material's surface.
Conclusions:
- The developed LTM effectively addresses challenges in low temperature embedding.
- This technique is advantageous for examining specifically oriented or laminated structures in electron microscopy.
- The method offers improved specimen preparation for high-resolution imaging.