TriM Scope overview - LaVision BioTec GmbH

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TriM Scope II - Introduction

The TriM Scope II is a modular 2-photon microscope that can be adapted to a various number of applications: from developmental biology, cancer research and immunology to neuroscience. Thereby, different types of measurements from 3D timelapse experiments to fast calcium imaging are possible.


TriM Scope II – Overview
The TriM Scope II is a modular 2-photon microscope platform that can be adapted to a various number of applications: from developmental biology, cancer research and immunology to neuroscience. Thereby, different types of measurements are possible: from 3D timelapse experiments over hours, large scanfields, fast 3D stack recording and fast line scanning to calcium imaging. 

Potential measurements
- Fast intravital imaging with multiplexed beams and resonant scanner   
- FRAP, photo-activation / -stimulation, uncaging, optogenetics, and line scanning  
- SHG and THG imaging with OPO and Ti:Sa laser
- Integrated FLIM imaging with LaVision BioTec’s FLIM detector
- Ultrafast 64 beam imaging with up to 100 frames/s @ 2560x2160 pixel

Benefits for 2-photon microscopy
- Excitation of red dyes
- Reduced phototoxicity and bleaching
- High imaging depth with 1100 - 1600 nm @ 200 fs
- Optimized SHG and THG excitation and detection


The TriM Scope II
The TriM Scope II comes with an upright, inverted, or both stands (the Doubleheader system) and allows simultaneous scanning with up to two Ti:Sa lasers, one or two OPOs, and various visible laser lines.
To increase the imaging speed it can be equipped with LaVision BioTec‘s 64 Beam Splitter, LaVision BioTec‘s Cloud Scanner, and a resonant scanner. In addition to its imaging capabilities the TriM Scope II provides various line scanning and advanced photo-treatment modes that allow fast data acquisition, photo-stimulation, uncaging, and bleaching.
Finally the TriM Scope II can be combined with a variety of detectors: up to 2 imaging camera detectors for 64 beam imaging, up to 8 NDD PMT detectors, up to 3 descanned PMT detectors, and LaVision BioTec’s FLIM detector.
To correct for tissue induced aberrations the Adaptive Optics Module can be integrated into the TriM Scope II. All the TriM Scope II add ons, the resonant scanner, independent galvo scanners, beam multiplexing devices or the Adaptive Optics Module, can be directly integrated into the TriM Scope II setup but also as an upgrade at a later time point.

Fluorescence, second, and third harmonic imaging of B16/F10 melanoma cells collective invasion into mouse dermis. By courtesy of Bettina Weigelin and Peter Friedl, Radboud University Nijmegen


Hardware features of the TriMScope II
- Combination of up to 2 Ti:Sa + OPO
- Chirp compensation optional
- Two internal EOMs, up to 8 EOM or AOM in total
- Adjustment control with 4 Quadrant Diodes
- Variable telescope, large diameter optics
- Motorized tubelens
- Various number of detectors
- ROI scanning
- Line scanning (high speed single line, polygon scanning)
- Mulitpoint access, mixed scan modes
- Line averaging scans
- Mosaic scans for large scan fields
Advanced Photo-treatment
- Photo-treatment in single and multiple ROIs
- Laser cutting
- Local power adaption for imaging and treatment
- Visible and UV lasers for photo-activation and photo-switching

Extended infrared multi-photon imaging
Most standard 2-photon microscopes are equipped with Ti:Sapphire lasers that deliver NIR laser radiation in the spectral range between 680-1040 nm. Therefore, 2-photon microscopy is limited to blue, green, and yellow dyes or fluorescent proteins. Red dyes or red fluorescent proteins are almost excluded from this deep imaging technique. Exciting red dyes or red fluorescent proteins requires pulsed laser radiation in the far infrared (>1100 nm), which is delivered by OPOs. Moreover, OPOs and lasers in the range between 1100nm and 1300nm enables efficient SHG and THG measurements. LaVision BioTec’s spectral detector allows simultaneous SHG, THG, and fluorescence detection. In combination with the transmission port the detection of SHG and THG is even more efficient.  
New lasers now deliver the wavelength range of Ti:Sapphire and OPO lasers in one system. The InSight® laser of Spectra Physics and the Chameleon Discovery® laser of Coherent have one tunable beam from 680 nm to 1300 nm and a second fixed beam around 1040 nm. Because both beams are coupled into the TriM Scope II, simultaneous excitation of several dyes with two different wavelengths is possible.  

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