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DSU
Look Closer & See Clearer! Olympus Spinning Disk Confocal
The DSU (Disk Scanning Unit) offers cost efficient semi-confocal image observation with a cooled CCD camera and an arc lamp illumination source. The DSU disk contains a pattern of slits that creates a virtual pinhole as the disk spins at 3000 rpm. It removes out of focus blur, thereby enhancing resolution. This yields clear, continuous, optically cross-sectioned images. The DSU is compatible with both inverted and upright Olympus microscopes, including the motorised IX81 and BX61 making it especially suitable for automated Z-sectioning followed by post image processing like 3D reconstruction. The water-immersion BX61WI and BX51WI electrophysiology microscopes and the entire line of water immersion objectives are also supported.
DSU Overview
In conventional widefield fluorescence microscopy, the inside structure of a thick specimen (<20 μm) cannot be observed clearly because of the significant contribution of out-of-focus light from above and below the focal plane. The DSU makes it possible to reject out-of-focus light by placing a spinning disk with an alternating pattern of vertical and horizontal slits in the confocal plane of the microscope. The slit disk spins at 3,000 rpm creating virtual pinholes which have an effect similar to the pinholes used in confocal laser scanning microscopy (CLSM).
Putting a new spin on flexibility
Olympus works with its customers to develop microscopes, modules and systems to provide better solutions for all microscopy, imaging and analysis processes. As a result, all Olympus microscopes are designed as “optical benches” enabling a range of modules and peripherals to be added with great ease. This drive for flexibility has led to the development of the new Olympus disk-scanning unit (DSU), which provides excellent confocal optical sectioning and motorised operation, with the added versatility of an arc lamp excitation source. The slit disks themselves feature a unique, patent-pending design. As a result of this careful development, the Olympus DSU provides researchers with confocal images with excellent contrast and resolution compared to conventional widefield microscopy. By using arc lamp excitation sources, the Olympus DSU offers the maximum wavelength flexibility within an economical package.
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Widefield versus DSU image quality
Images from the same section of PtK2 cell microtubule fragments: Image on the left was acquired using standard, widefield fluorescence and image on the right was acquired using the DSU confocal. Note the excellent confocal effects of the DSU including the excellent optical sectioning and removal of out of focus light especially with thicker specimens.
Comparing 2-D images of cellular microtubule fragments taken with the DSU (Disk Scan Unit) versus standard, widefield fluorescence
Photos courtesy of:
OLYMPUS CORPORATION Scientific Equipment Division
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PtK2 cells: Conventional fluorescence microscopy
Excellent optical sectioning, superior resolution and removal of blur with disk-scanning confocal microscopy.
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A choice of disks for different purposes
Five different types of disks are available, with different slit widths to suit different objectives and specimen thicknesses. Exchanging one disk for another is easy using the provided tools. In the case of the BX61WI, the best confocal effect is obtained by selecting the appropriate disk to suit the objective in use.
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Easy operation
A simple hand switch close to the operator's hand makes it easy to control such operations as inserting/removing the confocal disk into the light path, exchanging cubes and opening/closing the shutter. It is quick and easy to exchange between general fluorescence observation and confocal observation.
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Advanced system performance
Confocal 3-D image stacks can be acquired easily using the Inverted IX81 or Upright BX61 microscope platforms with their built-in, precise Z motor. Image acquisition software can be used to control microscope, DSU and digital camera for a very easy to use complete solution to your imaging needs.
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Brilliant performance over a wide range
The DSU disk and optical components are UV-compatible and deliver outstanding performance from 350–700 nm, making it suitable for most available fluorochromes. The DSU system supports DAPI excitation in the near UV without modification.
Easy operation
Disk operation is motorised, allowing a computer to switch remotely between widefield and DSU modes. The system includes a motorised filter turret for the convenient change of fluorescence filter in DSU mode as well as a motorised filter wheel equipped with neutral density (ND) filters to maintain illumination intensity in different observation modes. The DSU can be combined with a motorised microscope such as the upright Olympus BX61 and the inverted Olympus IX81 for 3-D confocal imaging.
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High speed, high sensitivity – less phototoxicity
The system offers full-frame CCD image capture at up to 15 frames per second with a cooled CCD camera, making the DSU an excellent tool for live-cell applications where speed of acquisition and minimal phototoxicity is paramount.
Excellent cost-performance ratio
By utilising white-light sources, the DSU is a cost-attractive alternative to laser-based systems. A lamp house with standard mercury burner, for example, can be used in combination with a suitable set of filter cubes. Moreover, high-quality filter-mirror sets for GFP and RFP, a manual filter turret and a motorised filter wheel with neutral density (ND) filters are already included in the basic DSU system. Furthermore, as your needs change, you can easily update the fluorescence excitation characteristics by adding new filters and dichromatic mirror sets to coincide with your new fluorochromes. Laser-based systems, on the other hand, often require a new laser source as well as filters and mirror sets for each new fluorochrome.
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3-D extended focus
image obtained from a 230μm thick specimen via 70 discrete z-sections at 2μm increments. Objective: UPlanApo20X
Photo courtesy of: Prof. Kei Ito Dr. Takeshi Awasaki
Department of Molecular Biology Tokyo University
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ADVANCED IMAGING WITH Cell M & AND Cell R
By integrating the DSU into the versatile imaging system Cell M and the real-time imaging system Cell R, the DSU is an ideal device for optical sectioning in combination with a variety of imaging applications. Cell M and Cell R are specifically designed to meet the experimental requirements for multicolour fluorescence time-lapse image acquisition. Cell M’s system coordinator, a control board solely for controlling hardware, considerably increases imaging speed in comparison with systems driven by software alone. Synchronisation of all hardware and peripheral devices at microsecond accuracy is performed with the Cell R hyperprecision real-time control board, making the modular system particularly well suited for live-cell imaging and a broad range of life science experiments – including time-lapse imaging, multidimensional imaging, ratio imaging, FRET and TIRF microscopy and spectral unmixing. The intuitively structured Cell M and Cell R Experiment Manager is a user-friendly graphical drag-and-drop interface, making setting up even the most complex experiments exceptionally quick and easy.
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Complete light management with MT* illumination systems
For more comfort and more speed, the DSU is available with the superior multifunctional illumination systems MT10 and MT20 instead of a standard lamp house. The MT10 features an integrated 8-position excitation filter wheel (>85 ms), a shutter with less than 5 ms switching time and a 7-position attenuator to adjust light intensity without changing the colour temperature. The MT10 light source can be obtained as a stand-alone module or as an integral part of . For real-time imaging with , the MT20 offers more speed and more precision. A fast shutter (<1 ms) to avoid photobleaching when no image is acquired, an 8-position excitation filter wheel (>58 ms) and a 14-position attenuator make it the perfect tool for high-speed experiments.
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Highlights
- Image formation is obtained from a CCD camera that allows full frame images to be acquired at up to 15 frames per second. The DSU is excellent for live-cell applications where speed of acquisition and minimal phototoxicity is paramount.
- Disk control is fully motorized allowing a computer to easily engage the disk into the lightpath and select wavelengths via the included filter changer. Five disks are available of varying slit width and spacing allowing the DSU to be optimized for varying objective numerical apertures and specimen thicknesses.
- Fully motorized disk operation and wavelength selection allows computer control and easy switching between confocal and brightfield techniques.
- UV fluorescence excitation capable. DSU optical design is optimized for 350nm excitation allowing excellent performance from near UV to near IR. DSU accepts standard Olympus filter cubes for excellent system flexibility.
- A CCD camera is used for image acquisition allowing full frame, high resolution imaging at up to 15 f.p.s.
- Five disks are available of varying slit width and spacing allowing the user to optimize disk performance to different objective numerical apertures and specimen thicknesses.
- Confocal images can be collected with objectives from 10x to 100x.
- Disk switch-outs are easy to perform.
- DSU is powered by the microscope's power supply saving important bench space and maintaining cost effectiveness.
- DSU filter turret has six positions, and can accept up to 5 different fluorescence filter cubes for excellent system flexibility.
- Small Footprint: The DSU mounts off the back of the BX51, BX61 or IX2 conserving valuable bench space on either side of the microscope frame.
- Illumination system includes a field stop to control the area of the specimen that is illuminated.
| Spinning Disk Confocal Specifications |
| Confocal Scan Method |
Disc rotation method |
| Maximum scan speed |
Image acquisition less than 33msec/frame |
| Camera |
Recommended camera: CoolSNAP HQ, Orca ER |
| Excitation wavelength |
350nm - 700nm.
Wavelengths of less than 430nm at naked eye observation and DSU observation may reduce the confocal effects. |
Fluorescence wavelength
at observation |
At less than 450nm, use our HQ filter for observation |
| Observation mode |
Exchange between confocal and non-confocal modes can be performed through the software |
| ND filter for excitation |
An ND filter will be inserted automatically at the exchange of confocal and non-confocal modes |
Electromagnetic shutter
for excitation |
Can be controlled through the software |
| Microscope attachment |
Intermediate attachment method (other cameras can be mounted on C-mount intermediate attachment) |
Temperature
and humidity |
10°C - 35°C, 30 - 80% |
| Power |
Provided through microscope controller |
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