X-Light: Spinning Disk Confocal
Super resolution microscopy that answers the deep biological questions effortlessly
Based on a multi-spot structured illumination system, the DeepSIM is a reliable, simple to use and affordable solution to study
cellular structures up to an XY resolution of ~100nm.
Super-resolved optical sectioning, with Z resolution up to ~ 300nm, can be obtained using both high (60X – 100X) and low
magnification (20x – 40x) objectives to expand the range of applications to include complex 3D models such as tissues, organoids,
spheroids and small organisms.
The DeepSIM is designed to work with samples of thicknesses comparable to those used in confocal microscopy, giving super-resolved
data over 50µm Z in depth in non-clarified samples.
This means that more meaningful data can be obtained from native heterogeneous complex samples using routine preparation protocols.
DeepSIM enables the effortless study of live-cell dynamics through a temporal resolution greater than 10fps (1024×1024 px FOV),
allowing biological changes to be tracked at cellular and subcellular levels.
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A flexible, compact but full-fledged spinning-disk confocal solution, CICERO makes high-end fluorescence imaging accessible to every laboratory enabling a variety of applications.
CICERO is a complete widefield and spinning disk solution, it can be integrated into any imaging setup transforming it into a
user-friendly and reliable confocal system. Life sciences, metrology, and material sciences are among the disciplines relying
increasingly on high-resolution 3D imaging.
CICERO spinning disk solution delivers fast image acquisition speed (15K rpm) and sensitivity, easily enabling live cell imaging and
large-scale 3D object imaging. Due to its large field of view (up to 22mm FOV), CICERO offers a minimal scanning process and can
capture large samples in a single frame.
CICERO is a reliable and flexible solution compatible with both LED and Laser as illumination sources. Moreover, the wide spectral
range enables a large variety of applications. For this, CICERO allows to address both entry-level and challenging applications.
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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
BPAE Cells – Mitochondria, Actin and DNA stained
Hela Cells in Interphase-Cytoskeleton tubulin, DNA and cell cycle regulating protein
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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.
