A fluorescence imaging technique using a femtosecond, normally tuneable infrared laser to excite fluorescent dyes, multiphoton imaging allows structures deep within thick samples to be imaged, making it perfect for brain slice and in vivo imaging.
Multiphoton imaging finds application in visualising the smallest of structures, available through diffraction limited resolution and optical sectioning via the nonlinear excitation. The technique is also used to scan over a wide area to collect data from a number of cells for the investigation of more variables from one set of experiments.
Scientifica has improved on the common approach of 'one multiphoton system for all applications' by providing a range of multiphoton modules to complement its world renowned SliceScope microscopy system. The versatility of the Scientifica multiphoton system will appeal to researchers preferring a customised 'DIY' system, and to those demanding a complete system ready to image, thus keeping experimental downtime to a minimum.
Multiphoton imaging system should not only meet immediate experimental needs, but also be versatile enough to accommodate long term research plans. Scientifica's approach to multiphoton imaging is to provide the building blocks that can evolve in-line with these experimental needs.
In addition to creating hardware components suitable for a wide range of applications, the Multiphoton imaging software solution is completely open source, allowing researchers to customise their imaging protocols even further by making use of ScanImage and HelioScan to control scanning and acquire images.
Scientifica offers a modular multiphoton imaging system that meets the needs of researchers looking for key components to speed up their 'DIY' development process or a fully operational system ready to image. By selecting Scientifica's open source, modular approach, any rig can be developed over time to meet specific criteria, justifying the significant hardware investment made upfront.
The latest developments provide new and current SliceScope users with an upgrade path to multiphoton imaging. The central components in Scientifica's system are the Multiphoton Detection Unit (MDU) and scan head, which will vastly increase the capabilities of existing systems.
- Multiphoton scan head
- SliceScope Frame
- Multiphoton Detection Unit above stage variant
- Multiphoton Detection Unit sub stage variant
- Objective slider kit or Motorised Objective Changer
Industry leading galvanometers from Cambridge Technology are employed to offer a highly versatile scan head for capturing images with high resolution and quality. The 3mm mirrors moving at a speed of 1 ms/line make it suitable for line scan and frame scan applications, ideal for imaging morphological and functional occurrences.
Galvanometers offer the benefit of variable scanning speeds, adjustable to suit each individual application. A reduced scan speed can achieve the same cell image morphology results while decreasing the sample's exposure to laser light, helping to reduce bleaching or tissue damage. High frame rates of approximately 15fps can also be achieved if the possible interactions between cells over a larger area by lowering the image resolution need to be monitored.
Key features and benefits of Scientifica’s multiphoton imaging system:
- Scan head achieves optimum scanning across the field of view for high resolution and fast scanning
- Enables imaging of extremely small structures and monitoring of fast-occurring biological processes
- Adjustable capture area using Galvo mirrors enables a specific region of interest to be selected, improving speed and allowing more data capture
- All optical elements coated for 700 – 1200 nm wavelengths, allowing the researcher to alter the excitation wavelength without any change in performance
- MDU is optimised for the collection of valuable photons by using customised optics, mechanics and noise-reducing electronics
- Custom designed, extra-large fixed position collection lens sited very close to the objective back aperture to improve image quality and prevent shading in the corners of the image
- Fixed distance between the collection lens and objective back aperture enables optimal collection
- High signal-to-noise ratio achieved thanks to the custom preamplifier and high-voltage power supply
- Exchange of emission filters and dichroic made easy with the MDU
- Wide range of objectives and condensers can be used as collection lenses, maximising light collection and allowing brighter signals to be achieved MDU fitted with electronic safety features and shutter to prevent PMT damage, prolonging the performance and life-time of the PMT