X-Light V2tp

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.

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

L-FOV: Large Field of View

X-Light V2 L-FOV is the X-Light V2 variant which covers the latest very large field of view cameras introduced in the biological market.

Specifically customized on the Nikon Instruments Eclipse Ti2 microscope, it guarantees illumination uniformity, high-speed confocal acquisition and high optical imaging quality in a 25mm field of view.

X-Light V2 L-FOV delivers:

• Up to 25mm Field of View
• Illumination uniformity: drop-off corner to center <15%
• Confocal resolution with 60x NA 1.4 oil immersion Objective: <650nm
• Adapter mount for Eclipse Ti2 side port
• F-mount for camera mount

Brochure in PDF Format

Technical Specifications for X-Light V2tp

Feature	Detail
Disk Configurations Available	Dual Pinhole Patterns, for flexibility in matching smaller detectors with a single disk
	Single Pinhole Pattern, for maximum field of view
Acquisition Modes	Widefield, with fast motorized disk‐in, disk‐out
	Confocal, for disk with single pinhole pattern
	Confocal, for each pattern of dual‐pattern disk
Detector Field of View	Dual‐pattern disk: 10×10 mm (14 mm diagonal)
	Single‐pattern disk: up to 22 mm diagonal
Pattern Configuration	CrestOptics proprietary disk pattern design for high confocal resolution, improved rejection of Out‐of‐Focus information and higher S/N
	Confocal Resolution with 60×/1.40 NA oil immersion objective: <650nm
	40‐micron pinholes
	50‐micron pinholes
	60‐micron pinholes
	70‐micron pinholes
	Custom spiral patterns available upon request
Custom relay lenses	Diffraction‐limited across entire field of view when using high‐magnification high-N.A. objectives or low‐magnification low-N.A. objectives
Spinning Disk Rotation Speed	15,000 RPM
Excitation mount	Gimbal mount, for easy alignment with optical axis of microscope and for best S/N
Dichroic Filter Wheel	5 positions, motorized
Emission Filter Wheel	8 positions, motorized
Ease of Installation	Plug‐in Spinning Disk
	Includes tools for insertion and extraction of dichroic filters and emission filters
	Provides for focusing of detector focal plane without moving the detector
	Includes adapter for your choice of excitation: multimode laser or LED SMA 905 fiber

Brochure in PDF Format

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

• Standard and custom hole pattern
  • Patterns differ from one other in hole diameter and spacing, best choice depends on microscope configuration and on sample characteristics
• Two patterns on one physical mask
• Pre‐aligned module with integrated LED or LASER launcher
• Bypass mode for widefield illumination and acquisition
• Up to 18 mm field of view
• 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
• Lateral (X–Y) resolution ca. 120 nm measured
  • limited by properties of optics, by objective magnification, and by camera pixel size
• Axial (Z) resolution ca. 300 nm
• Compatible objectives: highly-corrected immersion 60x and 100x
• 3‐D axial range: tested up to 110 micron
• Time resolution: ~5 sec/frame calculation included
• Camera requirements
  • 6.5 µm pixel recommended for sCMOS camera
  • sCMOS camera; QE>80% and low readout noise ≤ 1e- strongly recommended

Video card/CUDA Requirements

• Recommended for Full Performance:
  • Compute Capability: Version 5.0 or higher
  • GPU Memory: 4 GB or more

VCS/SCSR Pixel Calibration Table

	Camera Pixel Size
Magnification	6.5 µm	11 µm	13 µm
60×	54.2 nm per pixel	91.6 nm per pixel	108 nm per pixel
100×	32.5 nm per pixel	55 nm per pixel	65 nm per pixel

Brochure in PDF Format