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Microscopy Systems

X-Light: Spinning Disk Confocal

CrestOptics X-Light V2
CrestOptics X-Light V3

X-Light V3 – Spinning Disk Confocal System

The next generation of X-Light spinning disk confocal series.

It relies on the cutting-edge technology, advanced optical design approach and engineering solutions developed by CrestOptics to meet the very high-end specifications required by most of the modern fluorescence microscopy applications.

The spinning disk design together with the careful design of the optical layout enhances sensitivity and image clarity.

The first confocal unit which allows dual camera imaging at the full field of view of 25mm on both cameras.

Key Features
  • Spectral range (confocal/widefield): 400-750nm
  • Fully automated experiments
  • Software controlled bypass mode: widefield to confocal
  • The fastest spinning disk in the market: 15.000 rpm rotation speed, up to kHz range
  • Choose your disk: 50um pinholes in two different versions based on different pinole spacing
  • Choose your disk: unique continuous spiral disk makes it faster for high-speed applications
  • User inspired: customize pinhole size and geometry on request
  • Exchangeable, synchronized and sealed: the disk box is one of the most versatile features of the new V3. Dust protection and synchronization signals out to the camera make it reliable and faster.
  • Lateral Resolution (FWHM): ~230 nm (High NA 1.4) diffraction limited
  • Axial Resolution (FWHM): ~600 nm (High NA 1.4)
  • High Confocal Quality: this advanced new series brings in a redesigned optical layout optimized to enhance sensitivity and signal-to-noise ratio in any dim and bright conditions
  • Ultra-large field of view imaging: maximum 25 mm
  • Dual-camera view: simultaneous dual-camera view up to 25 mm each camera
  • Software controlled filter cubes: 3 positions for simultaneous dual-color imaging
  • Illumination uniformity: microlenses based for high uniformity across the full field of view
  • Illumination FOV: different options to match different camera sizes and enhance efficiency
  • Illumination shaping: preserve your sample. Motorized aperture to cut illumination in the region surrounding the region of interest
  • Software controlled filter wheels: 4-positions automated cleanup filter wheel, 3-positions automated dichroic filter wheel, 8-positions automated emission filter wheel

Mouse microglia primary cells- Cytoskeleton microtubles and DNA

mouse microglia

BPAE Cells – Mitochondria, Actin and DNA stained

BPAE Cells - Mitochondria, Actin and DNA stained

Hela Cells in Interphase-Cytoskeleton tubulin, DNA and cell cycle regulating protein

Hela Cells
 

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CrestOptics X-Light V2
CrestOptics X-Light V2

X-Light V2 – Spinning Disk Confocal System


Confocal imaging system from Crest Optics

The X-Light V2 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 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

The X-Light V2 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 V2 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 V2 Confocal Imager is an extraordinary combination of capability and price.

Learn more about the Principles of Operation of the Confocal Spinning Disk.

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: 22×22mm field of view for large chips of scientific CMOS cameras;
  • Dual Hole Patterns: 12×12mm 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 V2 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.

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Below is a example using the X-Light Confocal System

Mouse Kidney axial sectioning
Three channels (Dapi, Alexa Fluor 488 WGA, Alexa Fluor 568 Phalloidin,) axial stack of Mouse Kidney sample. The video shows a comparison (not the same field of view) between Wide field, Confocal Spinning Disk and VCS super-resolution acquisitions of 8 microns kidney section. The total thickness of the sample is approximately 16 microns. The VCS reconstruction shows a much more clear axial localization of the nuclei (DAPI) within other cell structures with respect to the other techniques.