X-Light V2tp

Confocal imaging system from Crest Optics

The X-Light V2tp is a unique system for spinning disk confocal imaging. It features unparalleled versatility:

  • Compatibility with all major inverted and upright microscopes
  • Plug‐in spinning disk box, allowing a change of pinhole pattern in seconds
  • Use of custom relay lenses for mitigation of aberrations of of a large field of view
  • Utilization of full field of view in sCMOS cameras
  • Full compatibility with Crest Optics Video Confocal Super‐resolution module (“VCS”) for 3-D resolution enhancement
  • Movable disk for widefield imaging
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The X-Light V2tp Confocal Imager

The X-Light V2tp Confocal Imager is a full-spectrum spinning disk confocal imager that attaches to most major models of inverted or upright fluorescence microscope. It is designed to utilize the latest imaging technologies and is ideal for live cell imaging in biological applications.

The X-Light V2tp Confocal Imager allows imaging at the extremes of performance of a current‐generation scientific camera, and it provides for objectives used when imaging with a camera with a smaller detector. It takes advantage of laser launches and LED light sources.

The result is a flexible confocal add‐on for existing microscopes that brings the latest commercialized technologies to microscopy. The X-Light V2tp Confocal Imager is an extraordinary combination of capability and price.

Features

Acquisition Modes

A rapid motorized Disk In/DiskOut control provides access to widefield and confocal imaging modes. Switching between modes is automated by imaging software.

Nipkow Spinning Disk

A proprietary disk pattern design provides high confocal resolution, improved out‐of‐focus rejection and higher ratio of signal‐to‐noise. The result is < 800 nm confocal resolution with 60x 1.4 N.A. oil immersion objective.

Disks are available with pinholes sized for best results from different objectives:

  • 40–μm holes, for objectives with N.A. < 1;
  • 70–μm holes, for objectives with N.A. > 1;
  • Custom pinhole diameters are available.

Disks are available to best suit particular camera setups.

  • Single Hole Pattern: 22×22–μm field of view for large chips of scientific CMOS cameras;
  • Dual Hole Patterns: 12×12–μm field of view for each pattern, for flexibility for smaller chips of CCD cameras.
Motor

The disk rotates at 15,000 revolutions per minute, providing excitation and imaging at speeds of current cameras.

Illumination Subsystems
  • The excitation mount accomodates LED light sources or multimode fiber for lasers; its SMA connector provides high‐efficiency coupling;
  • The gimbal mount for excitation eases alignment to produce the highest ratio of signal to noise;
  • The motorized five‐position dichroic filter wheel and eight‐position emission filter wheel provide for automation and ease of interactive control.
Other optics

Custom relay lenses mitigate aberrations of of a large field of view.

Ease of installation and maintenance

The X-LIGHT V2tp addresses difficulties of both initial set-up and continuing maintenance of a confocal imager.

  • The gimbal mount for excitation eases alignment that is required for maximal ratio of signal to noise;
  • Adjustment of focus is accomplished without moving the camera;
  • Fitler extraction tools ease insertion and removal of dichroic filters and emission filters.

VCS: Video Confocal Super-resolution

The VCS (“Video Confocal Super‐resolution”) module is an add‐on for the X-Light V2.

Patented Approach with High‐Speed Processing

CrestOptics VCS enhances the 3‐D resolution of the standard light microscope. It performs a two‐dimensional scan of the sample with a pinhole pattern and applies multiple algorithms to process the the emission, producing the image. The VCS technology is patented.

To speed the extensive calculation required, the VCS module uses CUDA technology from NVidia. The result is very fast processing.

Multiple Modes of Operation

The addition of the VCS module to the X-Light V2 provides a confocal imager with three modes of operation: Widefield Mode, Confocal Mode, and Super-resolution Mode.

Principles of Operation with VCS

VCS (“Video Confocal Super‐resolution”) is based on narrow-field illumination using scanning patterns and wide-field collection of raw images.

Detection algorithms already developed are able to super-resolve 3‐D structures in both compact and sparse specimens.

Although other techniques proposed and industrially developed mainly dedicate their efforts in extracting information from a relatively low spatial frequency range, VCM detection methods harness non-linear calculations exploiting the tops more than the belly of the raw signal intensity, collected as a function of illumination and detection positional parameters.

The figure below shows the schematic diagram for a basic VCS microscopy system. In this set‐up, the light source is focused onto a specified pattern that is optically conjugated to the microscope conjugate focal plane through a relay system. The mask is moved in the optical path with an X‐Y piezo motor system at each camera acquisition. The emission is collected widefield, and extensive processing performed to produce the final image.

Optical Set-up of VCS Bypass Mode

The figure below shows a typical illumination pattern projected onto a sample.

Typical VCS Illumination Pattern Projected onto Sample
Typical VCS Illumination Pattern Projected onto Sample

Specifications for VCS (“Video Confocal Super‐resolution”)

  • LED and LASER source compatible
  • Piezo motors speed structured illumination scan
  • Motorized control for VCS pattern focusing and color correction
  • Motorized mask positioning from VCS mode to Widefield/Confocal mode
  • Maximum lateral resolution up to 115nm measured, limited by pixel size and depending on pattern, algorithm and number of sub frames
  • Compatible immersion objectives: 60x, 100x
    • Low‐magnification/high‐N.A. objectives are under test
  • 3‐D axial range: tested up to 110 micron
    • Limit is under test
  • Time resolution: ~5 sec/frame calculation included
  • Supported Cameras
    • EMCCD
    • sCMOS camera; QE>80% and low readout noise ≤ 1e- strongly recommended

Video card/CUDA Specifications

  • Minimum requirements
    • Compute Capability: Version 3.0
    • GPU Memory: 4 GB
  • Recommended for Full Performance:
    • Compute Capability: Version 5.0/5.2
    • GPU Memory: 4 GB or more

- Technical data sheet

Links

- X-Light V2tp with VCS by Crest Optics

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