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Fabian F. Voigt edited this page Jul 16, 2018
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Welcome to the mesoSPIM wiki! This wiki serves as documentation on how to set up, align and operate a mesoSPIM.
The mesoSPIM (mesoscale selective plane illumination microscopy) project aims at creating open-hardware microscopy platforms for imaging in cleared tissue. Core features of the current generation mesoSPIM setups are:
- compatibilty with all major clearing techniques including CLARITY and iDISCO
- large field-of-views, ranging from 2 to 20 mm using an Olympus MVX-10 macroscope
- dual-sided excitation
- optimized for fast screening of whole mouse brain samples, yielding relatively small datasets (10-14 GB/brain/color)
- near-isotropic sampling (5-10 µm in X,Y & Z) due to an axially swept lightsheet which leads to uniform z-resolution across the field-of-view
- modular sample holders and quick sample exchange
- large travel range (45 x 45 x 100 mm) to allow imaging of very large samples
Inspired by the openSPIM and openSPIN projects, the documentation for building and extending your own mesoSPIM is available here.
- You are looking for a versatile imaging platform for cleared tissue that can be tailored to your needs.
- Research groups and imaging facilities with experience in building and supporting custom microscopes.
- screening of large numbers of samples for clearing & labeling quality
- visualization of sparse cell populations or other structures (blood vessels, plaques)
- imaging live samples (this is the domain of classical light-sheet microscopes)
- high-resolution (<1 µm pixelsize) scans with large number of tiles across whole mouse brains (this is better done using light-sheet instruments not based on a zoom macroscopes such as the Zeiss Z.1., the COLM)
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Background
- mesoSPIM history
- Optical design
- Electronics
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Setting up a mesoSPIM
- First steps
- Preparing the software and electronics
- Preparing the microscope optics
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Setting the microscope up
- General alignment tips and tricks
- Installing the microscope base
- Setting up the detection path
- Alignment of the detection path
- Setup of the sample XYZ stages
- Setup of the excitation path
- Immersion cuvettes
- Set up a microscope config file
- Light-sheet co-alignment
- Set up initial ETL parameters
- Setting up lasers with the GUI
- Sample Handling
- Test Samples
- Troubleshooting
- Upgrades and custom variants