TriM Scope II

The modular 2-photon microscope

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 scan- fields, fast 3D stack recording and fast line scanning to calcium imaging.

About 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.

Collective invasion of B16/F10 melanoma cells into mouse dermis, detected by infrared multiphoton microscopy. Tumor cells expressing E2-Crimson and Histone-2B-GFP are (false-colored) yellow (cytoplasm) and orange (nuclei). Nerve fibers and fat cells are cyan (third harmonic) and collagen bundles and muscle fibers are green (second harmonic), respectively. Blood vessels and phagocytic cells are labeled with TM-RHodamine-dextran in red.
Bettina Weigelin and Peter Friedl, Radboud University Nijmegen

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.  

Collective invasion of MCA101 fibrosarcoma cells into mouse dermis, detected by infrared multiphoton microscopy. Tumor cells expressing Histone-2B-mCherry are (false-colored) green (nuclei). Nerve fibers and fat cells are gray (third harmonic) and collagen bundles are blue (second harmonic), respectively. Blood vessels and phagocytic cells are labeled with Al750 in red.
Bettina Weigelin and Peter Friedl, Radboud University Nijmegen

Publications

Mossakowski AA, Pohlan J, Niesner R, Radbruch H, et al. Tracking CNS and systemic sources of oxidative stress during the course of chronic neuroinflammation, Acta Neuropathol. (2015)

Radbruch H, Bremer D, Mothes R, Günther R, Rinnenthal JL, Pohlan J, Ulbricht C, Hauser AE, Niesner R, Intravital FRET: Probing Cellular and Tissue Function in Vivo, Int J Mol Sci., 21;16(5):11713-27 (2015)


Nobis M, McGhee EJ, Morton JP, Anderson KI, et al. Intravital FLIM-FRET imaging reveals dasatinib-induced spatial control of src in pancreatic cancer, Cancer Res., 73(15):4674-86 (2013)

Bakker GJ, Andresen V, Hoffman RM, Friedl P, Fluorescence lifetime microscopy of tumor cell invasion, drug delivery, and cytotoxicity. Methods Enzymol, 504:109-25 (2012)

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TriM Scope II

Simultaneous scanning

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 -stimu- lation, 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.