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Immuno correlative light and electron microscopy on Tokuyasu cryosections
Viola M J Oorschot1, Tamar E Sztal2, Robert J Bryson-Richardson2
1Monash Micro Imaging, Monash University, Melbourne, Victoria, Australia.
Methods in Cell Biology
|October 8, 2014
Summary
Correlative light- and immunoelectron microscopy (CLEM) on Tokuyasu cryosections enables researchers to find rare structures. This method uses fluorescence microscopy to locate targets and electron microscopy for high-resolution imaging.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biophysics
Background:
- Locating rare cellular structures using electron microscopy is challenging.
- Correlative light- and immunoelectron microscopy (CLEM) offers a solution by combining light and electron microscopy.
- Tokuyasu cryosections are essential for preserving cellular structures for CLEM.
Purpose of the Study:
- To provide a detailed guide for performing CLEM on Tokuyasu cryosections.
- To demonstrate the application of CLEM in identifying rare aggregates in a zebrafish myopathy model.
- To highlight the versatility of the CLEM method for various biological samples.
Main Methods:
- Sample embedding and cryosectioning using Tokuyasu method.
- Immunolabeling of fluorescently tagged structures.
- Correlative imaging using light microscopy and high-resolution electron microscopy.
Main Results:
- A comprehensive protocol for CLEM on Tokuyasu cryosections is presented.
- The method successfully identified and imaged eGFP-actin aggregates in zebrafish muscle fibers.
- CLEM enabled high-resolution visualization of rare structures within their cellular context.
Conclusions:
- CLEM on Tokuyasu cryosections is a powerful technique for locating and characterizing rare cellular structures.
- The described method is adaptable for diverse biological samples, including small organisms and tissues.
- This approach significantly enhances the efficiency of ultrastructural analysis in biological research.
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