- Includes Core Components to Build a Multiphoton Imaging System
- Non-Descanned Design
- Microscope-Less Design Enables Imaging of Large Samples
- Directly Compatible with Thorlabs' Cage System Components
- Scan Head with Galvo-Resonant Scanner Pair
- Near-Infrared Scan and Tube Lens
- Dichroic Mirrors/Emission Filter Blocks
- Dual High-Sensitivity PMTs
- Computer with ThorImageLS™ Acquisition Software and 24" Monitor
The MPM-2PKIT Multiphoton Essentials Kit is Thorlabs’ solution for researchers who desire to build their own multiphoton imaging system without sacrificing performance. This kit contains the core components necessary to easily configure a modular multiphoton imaging system for a specific application. Additionally, the microscope-less design enables imaging of large samples, such as whole animals, and reduces the overall footprint occupied on a workbench.
The MPM-2PKIT Essentials Kit is derived from the optical system of our MPM200-2 Multiphoton Imaging System but made available in component form. The kit includes a scan head, a near infrared (NIR) scan and tube lens assembly, a primary dichroic mirror block, a filter block that holds a secondary dichroic as well as emission filters, two high-sensitivity GaAsP PMTs, and image acquisition electronics and software.
Thorlabs also offers a complete turnkey multiphoton solution, our MPM200-2 Multiphoton Imaging System.
|Multiphoton Essentials Kit Beam Path|
|Two-Photon Essentials Kit Specifications|
|Wavelength Range||680 - 1400 nm|
|Objective Pupil Diameter||20 mm (Max)|
|Field-of-View||16 mm Diagonal Square (Max) at the Intermediate Plane|
|Scanner||X: 7.8 kHz Resonant Scanner|
Y: Galvonometric Scan Mirror
|Scan Speed||30 fps @ 512 x 512 pixels|
|Scan Zoom||1X - 8X (Approximate)|
|Scan Resolution||Up to 2048 x 2048 Bi-Directional Acquisition|
Up to 4096 x 4096 Uni-Directional Acquisition
|Scan Mode||Point X-Y Scan|
|Primary Dichroic||680 - 1600 nm Longpass|
|Beam Conditioner (Optional)||Variable Beam Expander (1X - 4X)|
Motorized Beam Attenuation (λ/2 Wave Plate and Polarizer)
Optional Dispersion Pre-Compensation
|Non-Descanned Detectors||2 Ultra-Sensitive GaAsP PMTs (Coupled to Primater Dichroic Block|
and Secondary Dichroic/Emission Filter Block)
|Wavelength Range||300 - 720 nm|
|Filter Cube||Single, User-Changeable; Dichroic: 480LP, EM1: 445/45, EM2: 530/43|
Deep Tissue Imaging
Multiphoton microscopy is capable of imaging at extremely deep tissue depths.The extended excitation range allows the use of lasers at wavelengths longer than 1μm. As a result, more photons are able to reach fluorophores deep within the sample, which improves maximum imaging depth. A full Field-of-View non-descanned detection scheme with GaAsP PMTs positioned directly behind the objective ensures more precious photons reach the detector by collecting signal photons scattered by the sample but still pass through the objective.
Live Cell Imaging
Multiphoton microscopy is well suited for live cell investigations where samples are imaged over long periods of time. High-speed scanning to reduce photobleaching during live cell imaging is therefore advantageous. Thorlabs' Multiphoton system achieves high-speed scanning with a galvo-resonant scanner pair. The use of NIR two-photon excitation facilitates deep penetration into the sample, minimizes photobleaching, and lowers phototoxicity as compared to single-photon excitation technologies. Multiphoton microscopy has been used successfully to track GFP-labeled cells and cellular structures such as G-protein receptors in C.Elegens (see photo to the right).
Multiphoton imaging of GFP fluorescence in dopamine receptors in C. Elegens. Olympus 20X 1.0W. Sample provided by William Ryu, University of Toronto.
Label-Free Imaging with Second Harmonic Generation
Through two- and three-photon excited fluorescence of endogenous fluorophores, sub-celluar structures can be observed without the use of externally applied fluorescent dyes. For example, two-photon excitation fluorescence images of NAD(P)H, retinol, or flavins can provide functional chemo-biological information or be used as structural markers. Second harmonic generation requires highly ordered structures that lack inversion symmetry. The most common constituent of biological tissue that satisfies these conditions is collagen which can provide information on the extracellular matrix. Third harmonic generation can be observed at boundary interfaces and is useful in observing lipid bodies.
Second harmonic generation from chicken tendon at the interface of the connective tissue around the collagen fiber bundle. Scale bar = 100 μm.
Theory of Multiphoton Microscopy
Multiphoton excitation occurs when two or more photons arrive at a fluorophore simultaneously whose sum energy satisfies the transition energy. A new photon is emitted when the flourophore relaxes to its ground state, after small non-ratiative losses. Second and third harmonic generation (SHG and THG respectively) occurs when the incident photons are simultaneously annihilated and a new photon of the summed energy is created.
Optically Thin Images in Thick Samples
Two-photon excitation and SHG are nonlinear processes where the signal generated is dependent on the square and cube of the laser intensity for three-photon excitation and THG. The nonlinear nature of signal generation requires high photon densities provided by focusing femtosecond pulses to a diffraction limited spot. The output signal generated is mostly confined to the focal plane of the objective. Therefore, optically thin images from within thick samples are obtained. Three dimensional reconstrctions are made by moving the objective focus deeper into the sample.
The ThorImageLS software is a powerful Windows®-based acquisition software. The easy-to-use GUI interface coordinates system control, image acquisition, and animation playback through communication with all the hardware components driving the Thorlabs Multiphoton Microscope. APIs for LabVIEWand MATLAB (ScanImage compatible) are also available.
- Flexible Framework of Peripheral Control
- Multiphoton Scan Head Control
- Z-Stepper Motor Control
- Photomultiplier Gain Control
- Excitation Laser Control (Power & Wavelength)
- Beam Size Control
- Real-Time Background and Flat Field Correction
- User Control of Detection Channels
- Flexible Image Size and Location Adjustment
- Save and Recall Experimental Settings in XML Format
- User-Selectable Color Assignments for Detection Channels
- Selectable Color Channels
- Up to Four Simultaneous Channels
- Real-Time Capture
- 30 Frames per Second (512 x 512 Pixels)
- Streaming to Disk
- Z Volume Capture
- User-Defined Time-Lapse
- Up to Five Dimensional Data Collection
- Hardware Triggering for Experimental Initiation
- Image Formats: JPEG, TIFF, and AVI Movies
- Experiment Playback
- Image Histograms
- Region of Interest (ROI) Measurements
- Line Profile Measurements
- AVI Movie Creator
- Compress Image Sequences into an Easy-to-Share and Publish Format
Software Development Kit
- Develop Custom Software for Full Control of Thorlabs' Multiphoton and Confocal Imaging Systems
- Example Code Included to Simplify Application Development
- Libraries Provided for C++ and LabVIEW Languages
- Available Upon Request