Advanced instrumentation
for Photomanipulation

Phasor

Vector

Ablate!

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 is 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

Ablate! photoablation of chromosomes in an anaphase spindle. U2OS cell line (human oseosarcoma) stably expressing mCherry-tubulin (red) and H2B-GFP (blue-green). Two 532nm Ablate! pulses (each ~1.3ns) with a 5ms delay between pulses and Vector movement from position 1 to position 2. Captured on Marianas with CSU-X spinning disk confocal, 1K EMCCD, 63x 1.4NA oil objective, mSwitcher, Ablate! and Vector. Image courtesy of Dr. Marin Barisic, Danish Cancer Society Research Center.

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