DeltaVision OMX brochure.indd

AppliedPrecision®
DeltaVision|OMX
3D-SIM Super-Resolution Imaging
DeltaVision|OMX image
immunogold TEM micrograph

What is super-resolution?
In conventional microscope systems, image resolution
is limited by the angle of light that can successfully
traverse the light path and enter the objective lens.
While objectives can be built with very high numerical
apertures, a limit is reached when light can no longer
cross the interfaces between different refractive indices.
At this point, Brewster’s angle is achieved and additional
light and information (resolution) do not make it into
the objective lens. This angle ultimately limits the
resolution of all microscope systems. Once this limit
is reached, a system is said to be diffraction limited.
Super-Resolution is loosely defined as the ability for an
imaging system to exceed this limit driven by physics
and the wavelength of light and provide resolutions
that are significantly better than the diffraction limit
described above.
Applied Precision has led the way over the past fifteen
years in empowering scientists to fully exploit this limit
with image restoration (deconvolution) microscopy
included in the DeltaVision Core and personalDV
Microscopy Systems improving resolution by
approximately 15-20% and contrast ten fold. In recent
years, new methods have been developed to surpass
the diffraction limit. These methods allow precise
visualization and measurement of features that are less
than one-half of the size of those seen with conventional
microscopy. Improvement of resolution by a factor
of 2 or more is how Applied Precision defines superresolution
microscopy.
How does DeltaVision OMX achieve
super-resolution?
DeltaVision OMX uses 3D-SIM super-resolution
technology developed by the labs of Drs. Sedat,
Gustafsson, and Agard at UCSF. DeltaVision OMX is
a 3D structured illumination microscopy system that
enables super resolution imaging. DeltaVision OMX
surpasses the 250nm resolution limit by a factor of two
in x, y and z, and enables imaging beyond the surface
of the coverslip with multiple probes. This technology
will resolve features previously invisible to traditional
microscopy and lets you image more of your biology,
not just the biology that falls within a fraction of a
micron of the basal surface of the cell.
The DeltaVision OMX technology can image five,
ten, fifteen, even twenty microns into cells and
tissues. DeltaVision OMX works with conventional
fluorochromes eliminating any need to genetically
engineer novel or complex photoswitchable probes
so that you can apply the preparation methods and
fluorescent labeling reagents (antibodies and protein
tags) currently used in the lab.

What is 3D-SIM?
DeltaVision OMX extends the resolution limits of
microscopy by superimposing a high-resolution
sinusoidal illumination pattern onto the sample. This
pattern generates moiré interference patterns with the
high-resolution information contained within your
sample. These moiré patterns occur at lower frequencies
than the original high-resolution information in your
sample. By translating and rotating this illumination
pattern around the numerical aperture, twice the
spatial information can be collected by the microscope
effectively doubling the resolution of the microscope.
standard widefield
What about Temporal
Resolution?
Super-resolution is currently available for imaging fixed
samples. However, DeltaVision OMX is also designed
to deliver superior live cell performance.
The unique optical path of the DeltaVision OMX
system enables true simultaneous imaging of up to
four channels without compromising image quality. By
combining this light path with novel timing control, all
aspects of the imaging train from illumination shutter
control to camera timing and position control systems
are precisely timed to deliver the fastest possible
performance.
How fast? Up to 65 frames-per-second in each of up
to four simultaneous channels; the equivalent of 260
frames-per-second. These frame rates are not limited
by 3D imaging; a stack of over 60 Z-sections can be
acquired in four simultaneous channels in less than one
second.
Applied Precision has a well-earned reputation as
the leader in engineered imaging solutions for high
performance microscopy. DeltaVision OMX is a fully
designed and integrated imaging solution that takes this
performance to new levels.
An established technology
DeltaVision OMX
Two isoforms of beta-tubulin in a cultured neuron - Image courtesy of Stefanie
Kaech Petrie and Aurelie Snyder, Advanced Light Microscopy Core at The
Jungers Institute Oregon Health & Sciences University
DeltaVision OMX systems have been in place
around the world since 2008. As scientists make
groundbreaking discoveries using 3D-SIM, the demand
for the technology has grown. DeltaVision OMX
super-resolution systems are in use at the following
institutions: The University of California, Davis, The
Oxford University, The Samuel Lunenfeld Research
Institute, The Georgia Institute of Technology, The
University of Dundee and The University of Illinois. In
2009, the number of research facilities using DeltaVision
OMX grew to include Cold Spring Harbor Laboratories,
Ludwig Maximilian University of Munich, University
of Technology Sydney, Massachusetts Institute of
Technology and University of Cambridge.
DeltaVision|OMX

DeltaVision|OMX
What does it take?
This kind of spatial and temporal resolution can not
be achieved without careful engineering and testing. It
starts with meticulous system design. Each component
is carefully chosen and selected to deliver the highest
performance possible with today’s technology. Each
component is scrutinized for maximum performance:
• API certified highest quality PSF objective lens
• Custom high-performance reverse light-path beam
splitters
• Custom detectors
• Custom laser designs
• Proprietary motion control
• Proprietary timing control
• Proprietary light scrambling methods
• Proprietary light structure generation
• Fully optimized microscopy chamber
• Fully integrated laser table
• Integrated electronics
• Ergonomic workstation design
The unique stage design of the DeltaVision OMX renders the best stability and
repeatability available (lateral and axial stability images
standard widefield
DeltaVision OMX
Hippocampal neurons - Image courtesy of Eric Dent, University of Wisconsin Madison

How does 3D-SIM work?
The resolution of the microscope is limited by the amount
of spatial information that can pass sucessfully through
the optics. This is represented by the fine grid pattern
in panel 1. If we mix that high-resolution information
with a known signal that we can resolve (panel 2), we
generate a new pattern, the moiré pattern seen in panel
3. Here the pattern that we see is the difference between
the two patterns and can easily be represented without
high-resolution methods.
Likewise, in 3D-SIM, a three-dimensional illumination
pattern is superimposed on the sample. This pattern
generates a new pattern (a moiré pattern) that contains
both the illumination and sample data. By carefully
reconstructing the sample data from the moiré pattern, it
is possible to create a super-resolution three-dimensional
image of the original sample.
Because the illumination pattern is three-dimensional,
the image that is created contains both the lateral
(2D) and the axial (3D) data. This is limited only by
how deep into the sample the illumination pattern can
be maintained before light scattering attenuates the
illumination pattern (usually tens of microns into the
sample). This illumination pattern can be efficiently
generated with multiple excitation wavelengths allowing
multiple fluorochromes can be used in the same sample.
3D-SIM is the only super resolution method that can use
multiple probes in the same sample (currently up to 4)
and that can image well beyond the sample substrate.
panel 1
panel 2
panel 3
conventional
DeltaVision OMX
P53 Binding Protein 1 (P53BP1) at a double-stranded DNA break. Courtesy Gang Bao (Georgia Inst. Technology) and David Spector and Ileng Kurmaran (CSHL).

DeltaVision OMX Specifications
General Information
• Three-dimensional Imaging and Analysis System
• High spatial resolution beyond the diffraction limit
- ~100 nm lateral (depending on wavelength and optics)
- ~200 nm axial (depending on wavelengths and optics)
• Fast temporal resolution truly simultaneous live-cell
imaging
• Up to four-channels per data set
• Super resolution beyond the cover slip using 3D-SIM
Structured Illumination Microscopy (per technology
inventors Sedat, Agard and Gustafsson at UCSF)
• High speed imaging at up to 65 frames/second in each of
four channels
• Applied Precision certified optics
• Includes personalDV widefield deconvolution system for
integrated target ID and standard widefield imaging
• Custom Applied Precision transilluminator with ultrawhite
LED
• Choice of large field-of-view or high-speed camera options
• Laser fluorescence illumination
• Conventional and Structured Illumination light paths
• Image acquisition software
• Full suite of image processing and analysis tools
• Stage mapping between personalDV and
DeltaVision OMX systems
• Proprietary image restructuring software
• Time-lapse
• 3-D Modeling and analysis
• Includes toolkit, calibration slides, alignment optics, and
immersion oil kits.
Imaging Components
• Custom optics assembly with novel optical light path
• API certified highest quality PSF objective lens
• High precision, repeatable X, Y, Z stage
- Absolute accuracy: +/- 0.02%
- Repeatability: +/- 10 nm (Z), +/- 150 nm (X, Y)
- Step resolution: 5 nm (Z), 10 nm (X, Y)
- Maximum travel: 25 mm (X, Y, and Z)
• Novel Z-axis closed-loop piezo crystal drive
• Closed loop diffraction grating motion and rotary stage
• Fixed optics light path
• Integrated proprietary FPGA timing architecture
• Integrated motion control system
• Integrated camera control architecture
• Fixed-cell filter set with custom lasers
• Optional Live-cell filter set
• Includes 405 nm and 488 nm high power lasers
• Optional Lasers
- 514 nm
- 592.5 nm
- 642 nm
• High-speed excitation shutter for each laser (2 ms)
• Neutral Density filter wheel for each laser
• Novel widefield scrambling technology
• Olympus Plan-Apo 60X, 1.42 NA PSF “A” quality
objective lens (Standard)
- API certified for optimal 3D imaging performance
- UIS-2 extended color correction
• Optional U-Apo 60X, 1.30 NA Silicone Oil Lens
optimized for live-cell imaging
- API certified for optimal 3D imaging performance
- UIS-2 extended color correction
• High-performance custom 20 MHz cooled interline CCD
camera for maximum field-of-view
• Optional high-sensitivity custom EMCCD frame-transfer
camera for maximum sensitivity and frame speed
• OMX Master control computer
• Separate computers for each camera
• High-performance image processing and analysis
workstation
• High-performance dedicated gigabit Ethernet switch
Filter Sets
• Standard fixed-cell filter set (DAPI, Alexa488, Alexa594,
Alexa642)
• Optional live-cell filter set (CFP, YFP, EGFP, mCherry,
DIC)
• Optional filter drawers available
CCD Cameras
• Custom monochrome 20 MHz camera with Sony ICX285
ER progressive scan CCD
• Custom monochrome 10 MHz camera with E2V backthinned,
frame-transfer, CCD camera. ~92% QE with
electron multiplication (Optional)
R. parkeri infected Cos7 cells - Image courtesy of Matt
Welch, University of California at Berkeley
Streptococcus cells - Image courtesy of Cynthia
Whitchurch, University of Technology, Sydney, Australia
Image courtesy of Renato Mortara, Universidade Federal
de São Paulo (UNIFESP), Sao Paulo, Brazil

personalDV System Specifications
Workstations
• OMX Master Controller
- Dual-processor, dual-core 64-bit Xeon workstation
- 4 GB DDR RAM
- 500 GB hard disk
- Dual gigabit Ethernet adapters
- Windows XP-64 Professional
- 24” widescreen (1920 x 1600) flat panel monitor
• OMX Image Processing Workstation
- Dual-processor, quad-core 64-bit Intel Xeon workstation
- 4 GB DDR RAM
- 320 GB System hard disk
- 3 X 500 GB RAID 5 hard disk (1 TB total)
- Dual gigabit Ethernet adapters
- CentOS EL 4.0 Linux®
- 24” widescreen (1920 x 1600) flat panel monitor
• OMX Camera Workstations
- Dedicated to each camera
- Gigabit Ethernet
• OMX Instrument Control
- Proprietary Nanomotion-3 motion control interface
- Proprietary Applied Precision Field Programmable Gate
Array (FPGA) timing controller
softWoRx Image Processing and Analysis Software
• 5-D Image Window (X, Y, Z, Time, Channel)
• Multi-processor enabled software tools
• Multi-threaded image processing
- Deconvolution
- Volume rendering
• System-level processing queueing
- Deconvolution
- Volume rendering
- Quick projections
- TIFF file conversion
- Movie conversion (AVI and Quicktime®)
• Task Builder
• Orthogonal Viewer
• Arbitrary line profiles
• On-the-fly color blending
• Co-localization measurements (2 methods)
• File chooser with interactive thumbnails
• Time projections
• Data extraction tools
• Interactive volume rendering
• FRET analysis tool
• Drag-and-drop deconvolution queueing
• Support for file sizes over 2 GB
• Fully network compatible
• Image deconvolution methods:
- Constrained Iterative Algorithms (four methods)
- Nearest Neighbor
• Image import formats supported:
- Tagged Image File Format (TIFF®)
- UIC MetaMorph® STK
- BioRad® MRC-600 Pic
- Inovision ISee
System Configuration
• API Solid-state Transmitted White Light Illuminator
• InsightSSI 4 channel solid state illumination unit
• Fast shutter
• 10-position ND filter wheel
• 10-position EX filter wheel
• 6-position EM filter wheel
• Optical fiber
• Fiber Optic Module (FOM)
• IX-71 microscope base
• Custom DeltaVision Quad filter set
• 60x objective/1.42NA
• Stackable IC/MIC
• Dual processor tower computer, 400GB
• 25 mm stage
• CoolSnap ES2 Camera (PCI)
• Benchtop microscopy isolation platform
• 19” monitor
• Backlit keyboard, laser mouse
Stage Specifications
• Z axis
- Travel: 1 mm
- Resolution: 40 nm
- Linearity error:

DeltaVision OMX Specifications (continued)
• Image output formats:
- MPEG movie file export format supported
- Tagged Image File Format (TIFF)
- Quicktime ® movie file
- Photoshop ® (.psd) multi-layer file export format
• 3D modeling from 2D image data volume
• Per channel image analysis:
- Image stitch
- Image rotation on X, Y or Z axis
- Volume View
- 2D and 3D measurements
- Histogram review and non-destructive min/max intensity
adjustments
- Numerical data inspection from region of interest with
ASCII export
- 3D data graph from region of interest
• Complete Optical Transfer Function (OTF) Library for
common microscope objectives
• Archive data via network connection or on-board
DVD-RW
softWoRx Licenses
- Includes softWoRx DMS license
- Includes softWoRx Suite license
Ergonomic Design
• Optics Module
- Laser safety interlocks
- Light tight enclosure
- Vibration isolation (mechanical and acoustic)
- Class 100 HEPA filtering with positive pressure
- Vibration isolation table
- Integrated lighting
- 3 ft x 4 ft x 6ft (90 cm x 120 cm x 180 cm)
• Laser Module
- Laser safety interlocks
- Rigid optical design
- Multiple light paths
- 2.5 ft x 4 ft x 4 ft (105 cm x 120 cm x 120 cm)
- Integrated electronics rack
• Ergonomically designed workstation desk
- 3 ft x 4ft x 2.5 ft (90 cm x 120 cm x 105 cm)
• Maximum system weight = 1500 lbs (680 kg)
Electrical/Environmental Specifications
The DeltaVision OMX requires a laboratory roughly 8’x 16’
(2.5m x 5m) to provide adequate space for all the components
and to accommodate the work flow. The following are the
room requirements:
• Minimum Size: 8 ft x 16 ft x 8 ft high
• Dry, clean air or nitrogen lines:
- 100 psi regulated for clean gun, de-ionized
- 100 psi regulated for OMX table
• Room temperature maintained at 72° F (+/- 2° F over 4
hour period). System heat load ~ 5 kW.
• Power
- System power: 200-240 V ± 10%, 50 Hz ± 5%, 20
Amperes, single outlet located within 6 feet of laser/
electronics cabinet
- Additional power: 120 V± 10%, 60 Hz ± 5% or 200-240
V ± 10%, 50 Hz ± 5%
- Installation category 2 (office power)
- Total power ~3-5 kW
- An Uninterruptible Power Source or power conditioner
supplied by user is recommended
Cover Image - PCNA localization in CHO cell nuclei. Used with
permission from Andrew Belmont, University of Illinois, Urbana-
Champaign.
personalDV Workstation Table
Optics Module Cabinet Laser Module Cabinet Workstation Table
Applied Precision, Inc
1040 12th Ave NW
Issaquah, WA 98027
Tel: 425.557.1000
www.appliedprecision.com
© 2009 Applied Precision, Inc. All Rights Reserved.
Rev B 122209
Applied Precision, DeltaVision, softWoRx and NanoMotion
are registered trademarks and DeltaVision|OMX and 3D-SIM
are trademarks of Applied Precision, Inc. All other trade
marks are property of their respective owners.