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  • Microscopy and Laser Imaging

    602

    ASOM

    Spectral Radar OCT

    Swept Source OCT

    Video-Rate Laser Scanning Microscope

    Swept Source Lasers

    OCT Components

    Laser Microscopy Optics

    Microscopy Tools

    www.thorlabs.com

    Video-Rate Confocal Camera

    Confocal Camera Features ■ Confocal Imaging at Video Rate ■ Attaches to Camera Ports of Lab Microscopes ■ Diffraction-Limited Resolution Over a Large Field of View ■ Compatible with Thorlabs’

    Cage and Lens Tube Systems ■ Fiber or Free-Space Coupled Input

    and Output ■ Numerous Filter and Detector

    Configurations; Fluorescence Imaging Ready

    ■ Cost Effective Confocal Imaging Solution

    Thorlabs is pleased to announce the Video-Rate Confocal Camera System. This is a full-featured confocal microscopy system, yet the optical head is packaged in a compact ready-to-use module that connects directly to the camera port of any standard microscope.

    This powerful combination of proven Thorlabs imaging technology enables a true confocal solution at a fraction of the cost of competing systems.

    Thorlabs’ Confocal Camera offers real-time confocal imaging in a customizable open platform. The fiber coupled design of this system ensures a spatially filtered input beam, which is essentially a perfect Gaussian. In addition, the two single mode fibers that are used to deliver the illuminating light as well as collect the backscattered signal replace the pinhole that is used in traditional confocal systems. The confocal arrangement of the fiber position rejects out-of-focus light thus creating a true confocal image. All the necessary components such as a laser diode source, photodetector, associated control electronics, computer, and software are included.

    Confocal Camera Specifications Imaging: ■ Resolution:1 1 µm ■ Frame Rate: 23 fps @ 800 (X) x 640 (Y) Pixels

    (2-Channel Detection) ■ 3D Acquisition:2 800 x 640 x 256 (X, Y, Z) Pixels for 11 s Electronics: ■ Analog Input: 2 Channels, 14 Bit, 100 MS/s* ■ Analog Output: 4 Channels, 12 Bit, 1 MS/s*, ±10 V ■ Digital I/O: 8 Ports Optical:3 ■ Output Power: ~2 mW ■ Beam Diameter: ~ 4.5 mm

    Hardware: ■ CPU:

    Intel® Processor ■ Memory: 2 GB Memory ■ Operating System: Windows® XP Professional, SP2 ■ Hard Drive: 250 GB SATA ■ Optical Drive: 16X DVD ± RW ■ Monitor: 19" LCD 1280 x 1024 1 Resolution specified using a RMS40X objective and 660 nm diode. Actual resolution will

    vary with objective used. 2 When a piezo stage is used. 3 Output power and beam diameter measured at exit port of VCM101H with no objective using

    a 660 nm laser diode light source (included).

    *MS/s= Megasamples per Second

    (5.94") 150.8 mm

    (3.11") 79.1 mm

    (26.32") 668.6 mm

    (1.14") 29.1 mm

    (7.38") 187.3 mm

    (11.71") 297.3 mm

    (2.86") 72.7 mm

    VCM101H Video-Rate Confocal Camera Schematic

    ITEM# $ £ € RMB DESCRIPTION

    VCM101H CALL CALL CALL CALL Video-Rate Laser Scanning Confocal Camera

    Thorlabs’ new VCM101H Video-Rate Confocal Camera Mounted on an Olympus BX41 Microscope.

  • ASOM

    Spectral Radar OCT

    Swept Source OCT

    Video-Rate Laser Scanning Microscope

    Swept Source Lasers

    OCT Components

    Laser Microscopy Optics

    Microscopy Tools

    Microscopy and Laser Imaging

    603www.thorlabs.com

    Combined OCT and Confocal Microscope Solution: Multi-Modality Imaging on a Single Platform

    OCT and Confocal Synergies Shared Strengths: Video Rates

    In-Vivo Imaging 3D Imaging

    Complementary Strengths: OCT: ■ Large Field of View ■ Greater Depth Scanning Profiles

    Confocal Imaging: ■ Very High Resolution (Submicron) ■ Subcellular Imaging Capabilities

    The substantial line of OCT systems and solutions offered by Thorlabs demonstrates our commitment to developing OCT-based solutions to challenging imaging problems (see pages 2-15). In an ongoing effort to make this technology widely available, Thorlabs has recently developed an OCT Microscope Module that can be easily integrated (as an add-on component) with laboratory research microscopes (see page 9). That module is shown here mounted between the eyepiece and the objective turret of an Olympus BX41 microscope. For even greater versatility, this new system can be combined with the Thorlabs VCM101H Confocal Camera. By simply sliding a channel selector bar (See Fig. 1), this microscope system can switch among OCT, confocal, and standard microscopy configurations.

    Never before has such a powerful tool been combined so effortlessly into a commercially available laboratory microscope.

    OCT and confocal imaging are ideally suited to be combined into a single imaging system. Each imaging method brings unique strengths to challenging imaging applications. Since they share a number of features, it is possible for the two methods to work together seamlessly. Each technique has the ability to image at video rates, to test in-vivo sampling, and to obtain 3D images; these similarities allow the two methods to be used together to take advantage of their unique strengths. OCT systems allow for large- depth probing into samples and create images characterized by large fields of view (FOV). In particular, swept source OCT systems have a FOV up to 10 mm and can probe sample depths

    to 3 mm. In contrast, confocal imaging is beneficial because it produces very high resolution (submicron) images even though the FOV for an image is typically ~1 µm. By combining these two techniques, it is possible to obtain wide fields of view and resolutions capable of probing inside biological cells.

    Fig. 1 Swept Source OCT Microscope Module

    Combined OCT and Confocal Microscope ■ Thorlabs VCM101H Confocal Camera ■ Thorlabs Swept Source OCT Microscope Probe ■ Olympus BX41 Microscope

    Features ■ Enables Full-Featured High-Speed

    and High-Resolution 3D OCT Imaging for Research Microscopes

    ■ Compact Design for an Integrated Solution

    ■ Multiple Imaging Modalities Combined into One Microscope

    ■ Easy Switching Among Modalities for Imaging the Same Sample

    ■ Highly Reconfigurable Design ■ Supports Future Integration of

    Multiphoton and Fluorescence Imaging Channels

  • Microscopy and Laser Imaging

    604

    ASOM

    Spectral Radar OCT

    Swept Source OCT

    Video-Rate Laser Scanning Microscope

    Swept Source Lasers

    OCT Components

    Laser Microscopy Optics

    Microscopy Tools

    www.thorlabs.com

    Reflection Confocal Imaging Using the VCM101

    Microelectronics

    Confocal scattering images of a memory chip taken with VCM101H using a 660 nm incoming light source. The backscattered light is coupled into the single mode fiber and then detected with a photodiode.

    75 µm 55 µm

    20 µm

    2.

    1. Confocal scattering image of a memory chip with XZ and YZ cross-sectional images taken with a 60X objective. The total image size is 75 x 55 µm.

    2. Confocal scattering image of a memory chip taken using a 100X objective with 3D projection. Each microcircuit is 2.5 µm wide. Total image size is 55 x 35 µm.

    3. Reconstructed 3D projection model of the confocal backscattering signal from a portion of the circuit

    of a microchip, obtained using a 100X objective.

    Figure 1

    +20 0

    -20

    1.

    The VCM101H is a highly versatile imaging system. The standard reflection mode allows surface imaging of reflective opaque material for microelectronics, material science, and surface studies (see images below) and can also provide optical slicing of semitransparent scattering samples, as demonstrated by the image to the right.

    Pseudo-color 3D projection and cross- sectional confocal scattering image of a green leaf obtained with VCM101H using a 60X objective. The image was taken using a fiber coupled 660 nm laser diode. (250 x 185 µm, Z-depth is 80 µm)

    250 µm

    185 µm

    80 µm

    Leaf

    3.

    110 µm

    150 µm

  • ASOM

    Spectral Radar OCT

    Swept Source OCT

    Video-Rate Laser Scanning Microscope

    Swept Source Lasers

    OCT Components

    Laser Microscopy Optics

    Microscopy Tools

    Microscopy and Laser Imaging

    605www.thorlabs.com

    Confocal Fluorescence Imaging Using the VCM101

    (optional)

    Figure 3 shows the VCM101H reconfigured for fluorescence imaging. By changing the beamsplitter to a dichroic filter in the standard VCM101H setup and adding the appropriate excitation and emission filters, the system becomes a powerful fluorescence imaging tool. In most situations, a PMT should also replace the fast photodetector.

    Figure 3

    Peach Worm These images represent pseudo-color confocal fluorescence images of a peach worm taken with the VCM101H using a 60X water immersion objective. A total of 256 Z-slices (0.3 µm step-size) were used to create the pseudo-3D projection shown below. The total 80 µm Z-scan is represented by the selection of data images shown in Figs. 1 – 8, which are individual Z-slices at 10 µm increments. Each individual image is 440 µm x 430 µm.

    1

    7

    65

    43

    2

    8

    The true power of confocal laser scanning microscopy is most often demonstrated in fluorescent samples where the confocal technique spatially separates the desired fluorescent signal from the out-of-focus background fluorescence, allowing optical sectioning along the Z-plane. Combin