Multiphoton microscopy is the method of choice when you want to observe cells in action deep in living tissue at subcellular resolution. Depending on what you want to find out, you have to look into the sample from above, below, or from the side. Some applications require a lot of space for sample support, stage-top incubators, or even virtual reality setups.
To facilitate your work, we have rethought the concept of upright and inverted configurations for multiphoton microscopy and developed a completely new microscope stand. With its open design, configurability, and 360° rotatable lens mount, the TriM Scope Matrix adapts to any experiment in intravital imaging and beyond.
With regard to the most common applications in immunology, immuno-oncology, and neurobiology, the TriM Scope Matrix features two types of stages, the Intravital Stage and the Compact Stage. This opens up different configuration options for optimal sample support during your intravital experiments.
In research fields like immunology or immuno-oncology, the study of dynamic biological interactions related to tissue metabolism, angiogenesis, tissue remodeling, immunity, and inflammation is essential. Multiphoton microscopy shows what is happening in vivo in real time. Discover the features and add-on modules, making the TriM Scope Matrix a powerful tool to observe pathogenic processes and host responses within the living organism.
Upright or inverted? Each configuration is optimally suited for particular organs or tissues. Thanks to the turnable objective lens of the TriM Scope Matrix, there is no need to commit to one configuration, because samples can be observed from all perspectives.
Multicolor multiphoton microscopy experiments try to capture all actors in complex biological processes. This involves excitation and detection of several distinct fluorophores simultaneously.
With the TriM Scope Matrix, up to six targets can be captured on separate color channels in only one measurement. The standard 4× detector port enables the use of up to four GaAsP PMT detectors in epi-direction. The 6× detector port allows the simultaneous use of up to six GaAsP, GaAs, and/or Bialkaline PMT detectors in epi-direction. Latest shutter technology protects your valuable PMT detectors when no image is acquired.
When examining cell dynamics and interactions, the focus is sometimes more on speed or sometimes more on high resolution. Of course, the TriM Scope Matrix covers both.
Certain cell dynamics and interactions must be scanned at ultra-fast speeds, which requires a resonant scanner. Other applications require flexibility in scan speed, scan geometry, and pixel-dwell time. These can all be obtained with a galvanometric scanner in a flexible way. The TriM Scope Matrix scan head allows an easy switch between ultra-fast resonant and high resolution galvanometric scanning – or a combination of both with dedicated laser beams on each scanner pair.
Faster, brighter, smoother: With the 4× Cloud Scanner, the excitation process is parallelized by distributing the laser power to four individual foci, thus increasing the number of fluorescence photons per voxel and per time without raising photodamage.
In multiphoton microscopy experiments, fluorescent labeling of living tissue can be done by using both genetic and chemical tools, such as transgenic mice expressing fluorescent proteins or intravenous delivery of fluorescent dyes. But multiphoton microscopy is capable of even more as it can detect several unlabeled structures too.
Background noise can be annoying, but may contain meaningful information. Unlabeled structures and molecules, such as NAD(P)H, FAD, collagen, or tyrosine can be detected with the TriM Scope Matrix using weak but identifiable endogenous fluorescence (autofluorescence). This allows mapping of their relative abundance and spatial distribution. These structures are easily accessible with infrared light and thus deliver direct insights into tissue morphology and physiological processes.
The fluorescence-lifetime imaging (FLIM) module for the TriM Scope Matrix is a quantitative analysis tool to study molecular dynamics in different tissue structures. It is not the intensity but the lifetime of the fluorophore that is used to create the image contrast in FLIM. The fluorescence lifetime is not only different for different fluorophores, it also depends on the molecular environment of the fluorophore. Thus, FLIM can be used as an indicator for pH, polarity, temperature, oxygen saturation, protein interaction, or chemical species concentration, providing insights into metabolism or cell activation. With the help of FLIM, tumor cells, for example, can be directly distinguished from healthy tissue. Ultimately, fluorophores with the same excitation and emission spectra can still be distinguished with FLIM by their different fluorescent lifetimes. Accordingly, FLIM can also be used to distinguish signals from fluorescent labels and background autofluorescence.
The most interesting phenomena often occur deeper in the tissue. In order to work as least intrusive as possible, the penetration depth of the excitation light is of crucial importance. The TriM Scope Matrix offers solutions for deep tissue imaging.
The optics of the TriM Scope Matrix are designed for wavelengths up to 1750 nm, making it the perfect tool for three-photon microscopy. Longer excitation wavelengths scatter less in biological tissues. This extends the penetration depth, reduces out-of-focus excitation, and increases the signal-to-noise ratio at unprecedented depths. Long-term intravital imaging of bone marrow helps to uncover crucial steps of hematopoiesis, stem-cell migration, and immunological memory. Imaging entire lymph node vasculatures in vivo or penetrating deeper regions of the lymph node than two-photon microscopy would help to reveal previously unknown immune cell behavior. Three-photon microscopy takes optical penetration-depth into bones or lymph nodes to a new level.
When going deeper into the tissue, there is a risk that aberrations of inhomogeneous samples affect image
quality. The Adaptive Optics module includes a deformable mirror device that corrects for aberrations to restore the focal spot deep in the specimen and thus increases the fluorescence signal-to-noise ratio and resolution.
In order to fully understand both normal nervous function as well as diseases, the neural processes behind it need to be studied within the living tissue. Multiphoton microscopy helps scientists to understand neural networks, from single synapses to entire circuits, and how changes in the neural networks relate to behavior. Learn about the TriM Scope Matrix's special features and optional modules that will support your challenges in neurobiology.
The Twin Scanner for resonant and galvanometric scanning allows to capture fast dynamics, but also to run arbitrary scan patterns, even simultaneously. While the galvanometric scanner directs the laser to selected cells to activate subcellular components, the resonance scanner images the sample to monitor neuronal activity of individual cells within a neuronal network by excitation of Ca2+ indicators.
The In Vivo Stabilization module gets the most out of imaging living rodents. This hardware- and software-based motion stabilization tool includes a piezo-driven tripod to move the objective in all directions while the images are recorded with the resonant scanner. Together with additional software correction algorithms (x and y), it prevents blurring due to heartbeat and breathing or motion artifacts that occur in behavioral experiments with awake rodents in virtual reality systems. The In Vivo Stabilization module preserves all valuable data because no cropping is required as is the case with standard post processing.
When it comes to capturing the smallest signals as in calcium imaging, the TriM Scope Matrix is perfectly suited for highly efficient light collection and detection.
The 4× Cloud Scanner is the module of choice for calcium imaging and long-term in vivo measurements. The excitation process is parallelized by distributing the laser power to four individual foci, increasing the number of fluorescence photons per voxel and per time without raising photodamage.
Thanks to its modular design, spacious construction, and interfaces, the TriM Scope Matrix can be adapted to a wide range of experiments at both the hardware and software levels.
The TriM Scope Matrix provides numerous interfaces for your individual advanced optics and software tools. An additional treatment port allows for direct routing of (additional) modulated light sources to the sample that bypasses the scanners. The large breadboard top in the high-rack construction enables direct setup of individual beam routings for scanned laser beams, as well as for descanned fluorescent signal. Finally, the ImSpector acquisition software comes with a Python interface to give you direct access to numerous components of the microscope hardware to individualize scan-routines and use online data processing for direct feedback control.
Not sure which features and modules you need?
Set up your individual configuration together with one of our multiphoton microscopy specialists.
The TriM Scope Matrix is not an off-the-shelf device. Due to its modular design and upgradability with special modules, it is our goal to provide you with the exact device for your needs. Find the specifications of all essential and optional features and modules here or download the TriM Scope Matrix brochure.
|Resonant scanner||Galvo scanner|
|Unidirectional scan||Bidirectional scan|
|Scan frequency||8 kHz||1.2 kHz||2.0 kHz|
|Frame rate||30 fps at 512×512 pixels||2.3 fps at 512×512 pixels||3.3 fps at 512×512 pixels|
|Scan field (with 20× objective)||>1000 µm diagonal||>1000 µm diagonal|
|Quantum efficiency at peak wavelength||40% at 580 nm||25% at 375 nm||18% at 480 nm|
|Spectral response||300–720 nm||230–870 nm||230–920 nm|
|Photocathode area||dia. = 5 mm||dia. = 8 mm||dia. = 8 mm|
|Sensitivity adjustment range||1:50||1:10,000||1:10,000|
|Front-illuminated sCMOS camera||Front-illuminated CMOS camera|
|Sensor format||4.2 megapixels||4.2 megapixels|
|Active pixels (w × h)||2048×2048||2048×2048|
|Pixel size||6.5 µm × 6.5 µm||5.5 µm × 5.5 µm|
|Sensor size||13.3 mm × 13.3 mm, 18.8 mm diagonal||11.26 mm × 11.26 mm , 15.9 mm diagonal|
|Pixel well depth||45,000 e-||13,500 e-|
|Pixel frame rate||40 fps (100 fps optional)||80 fps|
|Readout mode||Rolling shutter||Global shutter|
|Read noise [median] rolling shutter||2.1 e-||13e-|
|Interface||USB 3.1 (Camera Link optional)||USB 3.0|
|Wavelength (nm)||Models (examples)|
|Ultra-wide tunable||680–960, 960–1300, 1025||CRONUS laser|
|680–1300||Spectra Physics InSight® X3+, Coherent® Chameleon Discovery NX TPC|
|Ti:sapphire||690–1080||Coherent® Chameleon Ultra II, Spectra Physics MaiTai®|
|OPO||1010–1340||Coherent® Chameleon MPX (pumped with Ultra II)|
|OP(CP)A||1300–1700||Class 5 Photonics White Dwarf, Light Conversion® I-OPA, Spectra Physics® Spirit-NOPA®-VISIR|
|Fixed-wavelength femtosecond||780, 920, 1064||Coherent® Axon Series, Spark Lasers® Alcor Series|
Non-descanned detection options
|Two-inch detection pathway||Spectral detector|
|Channels||Up to 4 GaAsP PMTs simultaneously in epi-direction for up to 4 color channels||Up to 6 GaAsP, GaAs, and/or Bialkaline PMTs simultaneously in epi-direction for up to 6 color channels|
|Shutter||Intrinsic electronic shutter (gate circuit) protects the photomultiplier tube by shutting it down, e.g., during optical treatment with VIS-lasers||Optional fast mechanical shutter (< 1ms) to block scattered laser light, e.g., during optical treatment with VIS-lasers|
|Design||Equidistant mounts close to the objective’s back aperture for high and identical collection efficiency|
|Spectral separation||Exchangeable large dichroic mirrors (78 mm × 55 mm) and dielectric filters (dia. = 50 mm)||Exchangeable dichroic mirrors (36.0 mm × 25.5 mm) and dielectric filters (dia. = 25 mm)|
|Channels||Up to 2 GaAsP, GaAs, and/or Bialkaline PMTs in forward direction|
|Design||Located close to the back aperture of the condenser (air or high-NA oil-immersion) lens for high and identical collection efficiency|
|Spectral separation||Blocking filter to exclude laser light, dichroic mirror and dielectric filters to separate signals (e.g. SHG and THG)|
|Laser lines||488 nm, 561 nm, 639 nm (fiber coupled, others upon request)|
|Channels||Up to three descanned GaAsP, GaAs, and/or Bialkaline PMTs|
|Compact Stage (inverted)||Intravital Stage (upright)|
|Dimensions and design||opening: 160 mm × 110 mm (for k-frame inserts)||top plate: 537 mm × 450 mm (w × d)|
magnetic stainless steel, with mounts for sample holder and a 25 mm × 25 mm thread (M6) array
|Height z||-||190–270 mm|
|Maximum travel range||x: 115 mm, y: 75 mm||x: 50 mm, y: 50 mm, z: 80 (180) mm|
|Resolution xy||0.5 μm||0.078 μm|
|Repetition accuracy||1.5 μm||1.0 μm|
|Maximum speed||30 mm/s||up to 4.4 mm/s|
In Vivo Stabilization
|Compensation speed||Up to 250 frames/s|
|Response time||<2 ms|
|Travel range||dia. = 40 μm (lateral), 48 μm (axial)|
|Software||Complete integration into ImSpector with readout of actual and corrected position|
|Number of foci||4|
|Length of line patter (with 20× Objective Lens)||~2 μm|
|Time delay between foci||~100 ps|
|Wavelength range||720-1050 nm (up to 1300 nm upon request)|
|Aperture (diameter)||15 mm|
|Working frequency||Up to 1 kHz|