OptoSplit III

Triple Emission Image Splitter
Cairn Research Ltd.

The OptoSplit III extends the OptoSplit image splitter concept, adding an optional second beamsplitter to split the field into either two or three separate, spatially equivalent, channels, which can be displayed side by side on a single camera sensor. The simple and accessible design makes the OptoSplit III an excellent platform for alternative applications, such as pFRET or multi-depth imaging.

Features

Fast Simultaneous Imaging

Splitting is usually performed on the basis of wavelength or polarization, allowing applications with a requirement for simultaneous or high speed acquisition of multiple emission bands or polarization states. The simultaneous acquisition of up to three images offers a major benefit over manual or electronic filter changers, as there is no longer need to pause acquisition while the filter position is changed. This allows your camera to be operated in fast stream modes.

Software Implementation

Device drivers are included in most commercial imaging packages to assist registration and to allow real-time and off-line ratioing of fluorescence overlays. Alternatively, the OptoSplit III can be used with simple image capture software, processing carried out manually offline or using our own MicroManager and ImageJ drivers.

Applications

Simultaneous multi fluorescent probe imaging
Simultaneous phase contrast/ DIC and fluorescence
Simultaneous multi depth imaging (using independent lenses)
Förster Resonance Energy Transfer (FRET)
Polarization Förster Resonance Energy Transfer (pFRET)
Radiometric calcium, voltage and pH-imaging
Total Internal Reflection Fluorescence (TIRF)
Polarization studies (Anisotropy)
Spinning disk confocal microscopy
Single Plane Illumination Microscopy (SPIM)
Super resolution STORM / PALM / SIM
3D super resolution PALM/STORM (using cylindrical lenses)

Specifications

Compact design with integral C-mount input and output ports (optional T- or F-mount)
Simple and precise controls for image registration
Interchangeable filter/ dichroic holders
Bypass without removing from optical path
Rotating filter mount for polarization studies
1x or 1.3x magnification
Support for sensors up to 18.8mm diagonal (13.3 mm x 13.3 mm sensors)
Angled and flat auxiliary drop in for neutral density filtering, polarizers or chromatic correction
Supports 25 mm filters and standard dichroic sizes
40 mm diameter optics
425 nm to 875 nm coatings on all surfaces
Fixed or variable center fully adjustable rectangular mask to delimit region of interest

Downloads

Literature

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Kastantin M, Faulón Marruecos D, Grover N, Yu McLoughlin S, Schwartz DK and Kaar JL. (2017) Connecting Protein Conformation and Dynamics with Ligand-Receptor Binding Using Three-Color Förster Resonance Energy Transfer Tracking. J Am Chem Soc. Jul 26; 139(29):9937-9948.

Mahou P, Julien Vermot J, Beaurepaire E and Supatto W. (2015) Multiphoton light-sheet microscopy using wavelength mixing: fast multicolor imaging of the beating Zebrafish heart with low photobleaching. Proceedings Volume 9329, Multiphoton Microscopy in the Biomedical Sciences XV; 93290Z

Wessel AD, Gumalla M, Grosshans J and Schmidt CF. (2015) The Mechanical Properties of Early Drosophila Embryos Measured by High-Speed Video Microrheology. Biophys J. Apr 21; 108(8):1899-907.

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