Total Internal Reflection Fluorescence and Photomanipulation Module
Vector3 is a motorized, spinning TIRF illuminator with three key imaging modalities: TIRF, photomanipulation and widefield epifluorescence. Intelligent beam steering and optical design allow for all three imaging modes to be combined in one compact device. Vector3 offers an expansive TIRF field of view (FN20) designed for modern sCMOS cameras. A motorized scan lens corrects for sample height variation and ensures ideal TIRF illumination and photomanipulation spot size across the visible spectrum.
Comparison between TIRF (left), wide field episode-fluorescence (middle) and a composite of both (right). When in TIRF, only the bottom of the cells are visualized, leading to a large increase in signal to noise of the membrane and organelles interacting with the membrane.
All Optical Electrophysiology
Nouveau Phasor is designed for 3D 2-photon stimulation of neurons in vivo. Four distinct modalities enable flexible optogenetics studies involving organisms from C. elegans and Drosophila to zebrafish, mice and larger mammalian species. Nouveau Phasor is able to simultaneously illuminate multiple regions in a 3D volume with the use of SLM-based computer-generated holography.
awake animal behavior
2 minute 15 second recording of mouse brain calcium activity in vivo via cranial window expressing ChRmine and GCaMP6m. 16 stimulation events were performed for single-cell activation and pair-of-cells activation. All cells had an increase in calcium activity upon stimulation.
Scanning Photomanipulation
Vector2 is a diffraction-limited high speed X,Y scanner that accepts lasers from UV to IR. Vector allows interactive examination of living specimens with a rapid single-point seek time of 0.3ms and fast ROI scanning (1500 lines per second). Common uses of Vector2 include FRAP, photoactivation, photoablation and uncaging. mSwitcher enables simultaneous use with Spinning Disk Confocal for advanced multi-modal methods. Vector2 is also designed for high-speed scanning imaging techniques such as multiphoton imaging or laser scanning confocal microscopy
photobleaching
Mammalian cell with membrane labeled in green before bleaching and FRAP curve in SlideBook.
Laser Ablation System
Ablate! uses an attenuatable 355nm or 532nm pulse laser (>60µJ pulses at 200Hz) to inflict localized damage to a diffraction-limited spot in intracellular structures for precise laser ablation and laser wounding. Full software control with SlideBook features automated intensity control and allows users to define one or more regions of interest as single diffraction limited spots or as user-defined shapes.
Photoablation
Mammalian cell with membrane labeled in green before (left) and after ablation (right)
Holographic Photomanipulation
Phasor 1-Photon uses a phase-only spatial light modulator (SLM) designed specifically for patterned and 3D point photomanipulation in optical microscopy. Phasor is compact and modular, allowing direct attachment to a microscope imaging port. Illumination is provided by LaserStack through a fiber optic coupling. Phasor is controlled by SlideBook software to manage region specification, hologram generation, experimentally synchronized optical path switching and excitation light gating. This state of the art control enables sophisticated experiment design incorporating pattern photomanipulation, including FRAP, photoactivation, photoconversion, and photostimulation.
3D Photostimulation
3D illumination pattern (left) applied to a 3D specimen (right) to stimulate multiple regions simultaneously.
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Phasor
3i offers the first commercial implementation of a phase-only spatial light modulator (SLM) designed specifically for patterned and 3D point photomanipulation in optical microscopy. Phasor is compact and modular, allowing direct attachment to a microscope imaging port. Illumination is provided by the 3i LaserStack™ through a fiber optic coupling.
Phasor is controlled by 3i’s SlideBook™ software to manage region specification, hologram generation, experimentally synchronized optical path switching and excitation light gating. This state of the art control enables sophisticated experiment design incorporating pattern photomanipulation, including FRAP, photoactivation, photoconversion, and photostimulation.
Solid state laser illumination from 3i’s LaserStack with the addition of 3i’s FiberSwitcher™ makes it possible to use a single set of lasers for multiple techniques on one system, such as point-scanning photomanipulation, spinning disk confocal imaging, and TIRF illumination. FiberSwitcher’s sub-millisecond change speeds make it possible to design elaborate experiments that utilize a given set of lasers for a combination of imaging techniques at very high speeds.
Advantages of computer generated holography fluorescence microscopy
Efficient use of illumination intensity by redirecting a significant portion of the light to regions where photomanipulation is desired.
Superior optical sectioning for larger regions compared to approaches that use Gaussian beam illumination.
Different points of stimulation can be focused at different z planes.
Entire photomanipulation patterns can be axially displaced from the imaging plane of focus.
Vector
Vector™ is a diffraction-limited high speed X,Y scanner that accepts lasers from UV to IR. Vector allows interactive examination of living specimens with a rapid single-point seek time of 0.3ms and fast ROI scanning (1500 lines per second). Common uses of Vector include FRAP, photoactivation, photoablation and uncaging. mSwitcher enables simultaneous use with Spinning Disk Confocal for advanced multi-modal methods. Vector is also designed for high-speed scanning imaging techniques such as multiphoton imaging or laser scanning confocal microscopy.
Ablate!
Ablate!™ uses an attenuatable 355nm or 532nm pulse laser (>60µJ pulses at 200Hz) to inflict localized damage to a diffraction-limited spot in intracellular structures for precise laser ablation and laser wounding. Full software control with SlideBook features automated intensity control and allows users to define one or more regions of interest as single diffraction limited spots or as user-defined shapes.
Features:
355nm or 532nm pulse laser
Available in fixed-point and motorized versions
Direct coupling to the microscope for optimal power delivery
Simultaneous widefield or confocal imaging with notch dichroic filter sets
Full software control with SlideBook and Photomanipulation module
Applications:
Cellular damage
Tissue damage
Organelle ablation
DNA damage
Vascular tissue damage for laser injury model
