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microscopy systems by gyvwpsjkko

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									                microscopy
                 systems
microscopy
systems




www.andor.com
 laser microscopy:
Andor Revolution - A Family of Products



                              Microscope Platform                                     iQ multi-dimensional imaging
                              • Andor Revolution products                             software
                                operate with any infinity correct-                    • Optimized for EMCCD – tight
                                ed microscope platform,                                  system synchronization with
                                upright or inverted.                                     minimal photon loss
                              • Microscopes from Leica, Nikon,                        • Build protocols through accessi-
                                Olympus, TILL or Zeiss are sup-                          ble wizards
                                ported.                                               • Comprehensive processing,
                                                                                         analysis and multi-dimensional
                                                                                         visualization                                   Andor Revolution
Laser Confocal Spinning Disk                           Piezo Nano Positioning
                                                                                                                                         Andor Revolution provides a frame-
• Ideal for high contrast confocal                     • Stage and objective positioners                                                 work for our range of laser-illuminated
  imaging of fluorescently labelled                       - 100, 200 and 500µm travel
  live cells                                           • Objective - Physik Instrumente
                                                                                                                                         microscopy solutions, including spin-
• Microlens disk enhanced laser                           PIFOC                                                                          ning disk confocal, TIRFM and photo-
  throughput                                              P721 / P725 - 100 and 500 µm
• Andor-enhanced with Semrock                          • Resolution of 2nm in 100 µm
                                                                                                                                         bleach and activation modalities. Our
  dichroics and apochromat                             • Tightly synchronized to give >                                                  spinning disk, live cell confocal instru-
  projection optics.                                      30 full resolution z sections per
                                                          sec!
                                                                                                                                         ment, illustrated below, combines
                                                                                                                                         Andor’s award winning iXon EMCCD
                                                                                                                                         cameras, solid-state laser combiner
                                                                                                                                         and the renowned Yokogawa CSU-X1
                                                                                                                                         confocal scanner “inside”. Andor has
                                                                                                                                         established a global distribution agree-
                                                                                                                                         ment with Yokogawa Electric
                                                                                                                                         Corporation and harnessed our opto-
                                                                                                                                         electronics engineering and manufac-
                                                                                                                                         turing knowledge to create powerful
                                                                                                                                         live cell confocal solutions. Andor's
                                                                                                                                         direct market presence and unique
                                                                                                                                         solutions bring you unrivalled perform-
                                                                                                                                         ance, product understanding and sup-
                                                                                                                                         port.

                                                                                                                                         Revolution encompasses a range of
                                                                                                                                         components, both hardware and soft-
                         Emission Discrimination
                                                                                                                                         ware that fit seamlessly together creat-
                         • Multi position filter wheel                                                                                   ing a complete laser microscopy solu-
                         • Dual port camera adapter
                         • Two, three or four channel field
                                                                                                                                         tions. An open flexible component
                           splitter                                                                                                      focus also allows us to provide key
                                                                                                                                         pieces of hardware stand-alone. We
                                                                                                                                         want to tailor solutions matched to
                                                                                                                                         your needs.
                                                                                                                                         At the core of Revolution systems is
                                      iQ workstation                                                 Andor Laser Combiner                Andor iQ, a multi-dimensional imaging
                                      • Delivers maximum performance                                 • Co-lineate a selection of solid
                                         to your Revolution system                                     state and/or gas lasers           software which synchronizes iXon
                                      • Acquire and handles huge                                     • Active AOTF blanking provides     EMCCD cameras with the CSU-X1
                                         datasets with consummate ease                                 microsecond shuttering and
                                                                                                       intensity control for minimum     confocal spinning disk and other key
                                                                                                       exposure                          hardware components such as Piezo
                                                                                                     • Blanking and channel cross-talk
                                                                                                       > 80dB                            Z100 z-stage and our Laser Combiner
                                                                                                                                         with AOTF for rapid laser line selec-
        iXonEM+ Ultra-Sensitive EMCCD                                Precision Controller Unit
        Technology                                                   • Provides synchronization to all                                   tion.
        • Detection of extremely weak                                   Revolution components
           signals with optimal S/N and                              • Active blanking minimizes
           high resolution                                              specimen exposure
        • Significantly reduced photo-                               • Synchronized piezo drive
           bleahing and phototoxicity                                   delivers highest frame rates
        • 1000x1000 @ 30fps to 128x128
           @>500fps




Front cover image: Cultured hippocampal neuron expressing cortactin (red) and membrane targeted YFP (green).
Image captured with the Revolution XD system.
Courtesy of Drs Nicholas Frost and Thomas Blanpied, Blanpied Lab,Dept. Physiology, University of Maryland, Baltimore, MD.
microscopy systems products:
iXonEM+ providing single photon sensitivity
                    Model            Features

                    • DU-860         128 x 128 pixels; 24 x 24µm; QE >90% for
                                                                                     All iXon detectors benefit
                                     high speed applications e.g Ca2+ flux
                                                                                     from: a) minimized dark cur-
                                                                                     rent from unparalleled –90°C
                    • DU-885         1004 x 1002 pixels; 8 x 8µm; QE >60%,
                                                                                     TE cooling – especially criti-
                                     35 MHz readout, high resolution intra-cel-
                                                                                     cal for confocal imaging
                                     lular imaging
                                                                                     where experimental back-
                                                                                     grounds are minimal, b)
                    • DU-897         512 x 512 pixels; 16 x 16µm; QE >90%,
                                                                                     Industry fastest vertical shift
                                     the ultimate combination of sensitivity and
                                                                                     rates (< 2µs/row) -
                                     speed
                                                                                     Minimizes clock-induced
                    • DU-888                                                         charge (CIC)
                                     1024 x 1024 pixels; 13 x 13µm; QE >90%,
                                     resolution and exceptional sensitivity com-
                                     bined

csu-x1 confocal spinning disk unit
                    Features

                    • 1500-10000 rpm disk, giving 300 to more       • 2x excitation throughtput improvement
                      than 2000 fps frame rate
                                                                    • FWHM Z section <=1.2µm
                    • Easy synchronization of exposure to disk
                                                                    • Semrock dichroic and emission filter pair-
                      speed
                                                                      ing provides optimum SNR
                    • Manual and motorized filter changer
                                                                    • Filter wheel mounting for fluorescence
                      models available
                                                                      and DIC analyzer
                    • Optional bright field bypass module
                                                                    • Andor 10 position available

                    • High performance custom Semrock dichroic mirrors; quad, triple and dual line combinations
                    • Custom optics fitted to camera adapters for higher throughput, chromatic aberration and
                      background noise reduction


laser combiner and multi-port switch uniquely flexible
                    ALC                                             MPD

                    • Integral AOTF for laser selection and         • Multiple ports for all laser lines - CSU,
                      modulation (cross talk <80dB)                   FRAPPA and TIRF
                    • Unique Active blanking, synchronized to       • Delivers 100% of laser power to one of
                      camera-minimizes specimen exposure              three fiber outputs
                    • Compact 4U 19” rack mount enclosure           • Fast galvanometer to switch outputs in
                                                                      approximately 1 ms, with external TTL
                    • Quiet, cool, low power consumption
                                                                      control
                    • Long life solid state lasers with excellent
                                                                    • Good stability, typically ~ 0.5% over a 8
                      stability (typical +/- 0.4% pk to pk)           hour period (includes SS laser long term
                                                                      intensity fluctuations)
                    • ALC allows Gas laser integration
                      e.g. Ar ion or mixed gas                      • Switching between CSU imaging and
                                                                      FRAPPA takes <5 ms



                               www.andor.com
precision controller unit flexible expansion
                    PCU100                                            PCU NEOS

                    • Provides system synchronization                 • Up to 5x 16 bit analogue outputs for
                                                                        external AOTF control
                    • Houses AOTF driver for ALC coupling
                                                                      • NEOS or AAEO AOTF support
                    • Active blanking driver for AOTF shuttering
                                                                      • Active blanking driver for AOTF shuttering
                    • 16 bit Analogue output for direct piezo Z
                      control                                         • 8 digital inputs for sensing events
                    • 8 digital inputs and 8 digital outputs for      • 8 digital outputs for triggering and expan-
                      sensing and triggering external events            sion
                    • Enables Revolution to slave or master.          • Information available for third party soft-
                                                                        ware support
                    • All under iQ software control




motorized xy & z control piezo z / motorized xy
                    XY Stage                          Z Stage                         Z Objective

                    • Open and closed loop            • Piezo Z stage with 100,       • PI PIFOC P721-100µm or
                      stages for your preferred         200 and 400µm ranges            P725-400µm travel
                      microscope
                                                      • 1nm repeatability             • Up to 20 Z sections per
                    • Typical travel                                                    second
                                                      • 2-8 nm resolution
                      >100x75mm, with 0.02µm
                                                        (depends on range)            • 1.25nm resolution
                      resolution
                                                      • Up to 100 Z sections per      • Analogue or digital con-
                    • 30mm/sec travel speed
                                                        second                          trol
                    • Perform multi-field scans
                                                      • Adjustable holder for 3x1     • Oil and water objectives
                      for 6D imaging
                                                        slide and 35mm petri dish
                    • Create 4D mosaics using
                                                      • Optional Tokai Hit stage
                      iQ software
                                                        top incubator


filter wheels & splitters emission discrimination
                    TR-FW10                                           Field Splitter

                    • 10 position, ø25 filterwheel with 40ms          • Optosplit II and III uses 25 mm filters
                      switching time (unloaded)
                                                                      • Field splitting for simultaneous 2 or 3
                    • Matched with CSU or microscope                    color imaging
                      "C"coupler
                                                                      • Matched to CSU or microscope "C" cou-
                    • Infinity optical path for optimum filter per-     pler
                      formance
                                                                      • DualView and QuadView also supported
                    • External controller allows support of
                      Sutter Smartshutter
                    • Multi-wheel Sutter controllers available




                               www.andor.com
software optimized imaging software & choice
                                                         iQ                                                                iQ Workstation

                                                          Andor iQ is our flagship live cell imaging                       • Dell Precision workstation
                                                          software, designed with flexibility and power
                                                          in mind. iQ - image and quantify - occupies                      • Acquisition Bandwidth and System
                                                                                                                             Configuration
                                                          a central role in our Revolution product
                                                          range and provides optimized control of                          • ImageDisk removes dependency on RAM
                                                          Andor's award winning iXonEM+ EMCCD                                for Huge Datasets
                                                          cameras and automation hardware for a                            • Multi-Core Processors and Multi-thread-
                                                          range of Bioimaging applications.                                  ed software
                                                          Continuous development and improvement
                                                                                                                           • High performance graphics with OpenGL
                                                          ensures that Andor iQ represents a powerful
                                                          and flexible core for live-cell imaging sys-                     • High resolution, high dynamic range, fast
                                                          tems.                                                              response LCD Monitors
                                                                                                                           • PCI cards included in system


camera port adapters single and dual port
                                                         Single Port Coupler                                               Dual Port Coupler

                                                          • Magnification X = 1.0, 1.2, 1.5, 2.0 -                          • Magnification X = 1.0, 1.2, 1.5, 2.0 -
                                                            other by request                                                  other by request
                                                          • Compatible with filter wheel or CSU+filter                      • Compatible with 'C'/'CS' lenses,
                                                            wheel                                                             C-mount, filter wheel or CSU+filter wheel
                                                          Andor couplers are designed to ensure opti-                       • Magnification select in each arm
                                                          mum throughput, minimal aberrations, mag-                         • Mounting cassettes 'store adjustment'
                                                          nification options and flexible detection                           simplifying exchange and minimizing
                                                          configurations. Our couplers match our                              readjustment
                                                          CSU enhancements and can also be config-
                                                          ured with a broad range of imaging optics
                                                          including microscope C-mount, 'C' and
                                                          'CS' mount lenses.


microscopes platforms & auto-focus
                                                          Diverse Compatibility

                                                          Revolution products are compatible with modern infinity corrected microscopes from Leica,
                                                          Nikon, Olympus or Zeiss to meet your preferred configuration. Note: For laser safety a 100%
                                                          side port switch is required.
                                                          Microscopes need environmental control to maintain living conditions for mammalian cell
                                                          preparations. We recommend temperature and CO2 control incubators from LIS (Switzerland).
                                                          Thermal and mechanical factors can impact the observation of living specimens when image
                                                          focus is affected by drift or vibration. Therefore we recommend that systems are configured
                                                          on vibration isolation tables. An important recent innovation in microscope design is focus
                                                          drift correction (FDC). FDC corrects focus drift, which can severely affect time-lapse movies
                                                          and is especially important in confocal and TIRF imaging, where focus and illumination are
                                                          tightly constrained. Nikon and Olympus have introduced FDC solutions, which Andor iQ sup-
                                                          ports to provide long-term drift free imaging from Revolution live cell imaging solutions. The
                                                          inset shows the new Nikon Eclipse Ti-E motorized inverted microscope.


* CSR (Constant Spectral Resolution)= λ/FWHM. Hence Resolution = 0.04nm @ 200nm. The FWHM corresponds to 3 pixel width. † = measured with 1200 l/mm grating
                                                                        www.andor.com
FRAPPA Photo-Bleach and Activate
                                            Features

                                             FRAPPA - Our latest innovation in laser microscopy is named after an acronym of the tech-
                                             niques which it supports. Fluorescence recovery after photo bleach (FRAP) and Photo acti-
                                             vation (PA) are established imaging protocols in which a computer-steered laser beam is
                                             used to photo-bleach or photo-active a user-defined region in the specimen.

                                             Andor’s FRAPPA uses a dual galvanometer scan head to provide a computer-steered laser
                                             beam delivery system. It can be configured in line with a CSU and/or camera and operates
                                             in two modes.

                                                  • Pass-through mode – FRAPPA acts as an optical relay system with 1:1 imaging
                                                  • FRAP mode – galvanometers are aligned to steer and image laser light from a
                                                    single mode optical fiber onto the specimen in a point scanning mode.

                                             Under Andor iQ control, the user commands FRAPPA to bleach or activate regions of interest
                                             with user-defined times, laser lines and powers.Laser switching is tightly synchronized with
                                             FRAPPA modes of operation, using our proprietary laser combiner multi-port switch (MPS).



Revolution System Features
• Ultimate sensitivity from combination of iXonEM+ EMCCD and
  CSU-X1 spinning disk or TIRF illumination.
• Fast frame rates (up to 1000 sec -1) with perfect synchroniza-
  tion.
• Active laser blanking provides:
       - minimal photo bleaching - particularly vital for
         non-ratiometric dyes.
       - reduced phototoxicity of living cells.
• Study live specimens with reduced fluorophore concentra-
  tions or expression levels.
• Minimal perturbation of physiological events.
• Synchronization of electrophysiology and photolysis with the
  Precision Control unit.
• Rapid optical sectioning with Piezo Z100 for 4D imaging - up
  to 100 sections sec-1.
• Detection limit down to single GFP molecules in parallel at
  supra-video frame rates.
                                                                     Courtesy Dr Iain Johnson, Invitrogen, Eugene, OR. Cultured neurons were loaded with DiSBAC2(3), a
• Photo-bleaching and activation to study diffusion and trans-       voltage sensitive dye and visualized with the Revolution 488. Exposure time of the iXon 887-BV was
  port of labelled molecules.                                        100ms and EM Gain 200. The figure shows a film strip from Andor iQ of maximum intensity projection
                                                                     time series. Time point 4 is taken almost simultaneously with the addition of KCl, which depolarizes the
• Powerful visualization, image analysis and tracking tools in       cells and shows a near instantaneous rise in DiSBAC2(3) signal. ∆F of almost 45% is observed in this
                                                                     example. Imaged at the 3D Microscopy of Imaging Cells Course, Vancouver 2005.
  iQ software.
• Detect and track many individual vesicles in parallel.
• Align and merge optically split images in iQ.
• Powerful tools for FRET visualization and analysis.




                                                         www.andor.com
                                                        microscopy techniques:
                                                         Confocal Dual Spinning Disk
                                                                                                            Revolution's confocal dual spinning disk
                                                                                                            technology provides an ideal platform
                                                                                                            for high speed, high SNR imaging,
                                                                                                            with low bleach rate and photoxicity.

                                                                                                            • Isolated detection channel with
                                                                                                              extremely low background - optimal
                                                                                                              for EMCCD technology
                                                                                                            • Efficient illumination coupling via a
Microscopy Applications                                                                                       microlens array (~60%)
Revolution components and systems                                                                           • Low peak power by parallel
benefit from common features across                                                                           illumination of ~1000 scanning laser
applications. Whether in confocal,                                                                            points
photo-bleach/activation or TIRFM imag-                                                                      • Efficient parallel detection with
ing this means state of the art perform-                                                                      highest QE - 90% peak
ance, flexible component-oriented sup-                   Confocal Dual Spinning Disk                        • True broadband operation - excitation
ply and an open-minded approach to                                                                            400-650nm - detection 420-750nm
systems and technically demanding                        Also
                                                       Available                                            • CSU optimization includes:
solutions. As we continue to provide                     from       Live Cell Confocal Upgrades
wide-ranging support for key hardware                   Andor                                               - Apochromatic detector               chan-
components, we also try to increase                                 Andor’s Revolution range includes our     nel optics
your choice through expanding our                         laser spinning disk live cell confocal
                                                          microscopy systems utilizing the Yokogawa         - Focusing single and dual camera
software compatibility. We are actively                                                                       adapters
involved in making the bridge from iQ to                  CSU. We have created a range of upgrade
MatLab, Imaris or Image-J and Micro-                      solutions for Perkin Elmer UltraView and          - Semrock dichroic filters optimized for
Manager as smooth as possible.                            Visitech system users to bring a new lease of       selected laser lines
Applications include:                                     life to legacy systems.

• Cell Motility
                                                         TIRF
• Live Cell Confocal
                                                         Total internal reflection fluorescence microscopy (TIRFM) provides an unrivalled technique
• TIRF                                                   for the study of single molecules and membrane related cell function, including adhesion,
• FRET Imaging                                           motility, endo- and exo-cytosis.
                                                                                                                 In TIRFM the electromagnetic field
• Ion Signalling                                                                                                 of the TIR light extends into the
• Fluorescent Proteins                                                                                           sample beyond the interface by
                                                                                                                 100-200nm into the medium of
• Photo-bleaching and Activation                                                                                 lower refractive index. Since the
• Single Molecule                                                                                                field decreases exponentially
                                                                                                                 along the z-axis, only a very thin
• Comet Assay
                                                                                                                 section of the specimen under-
• FISH                                                                                                           goes fluorescence excitation.
                                                         Prism-type TIRFM (LHS) and objective-type TIRFM (RHS)   TIRFM produces extremely high
• Immunofluorescence
                                                                                                                 contrast, low background images,
• Bio Chemiluminescent                                   ideal for detection with our iXon EMCCD cameras and is a valuable tool in the study of
                                                         e.g. cellular membrane traffic, cytoskeleton dynamics and single molecule studies.

                                                         Andor Revolution components provide:

                                                         • Matched illuminator and detector for maximum performance
                                                         • Flexible fiber-coupled 400-650 nm excitation compatible with all TIRF platforms
                                                         • Multiple fiber outputs for confocal, TIRF and Photobleach
                                                         • Integral AOTF – microsecond wavelength, shuttering and intensity control
                                                         • Active blanking - precise synchronization and > 80 dB isolation


Fixed muntjac cells labelled for actin and mitochon-
dria, illustrating extended focus montage imaging
with the Revolution XD confocal system.
microscopy techniques:
FRAPPA
Fluorescence Recovery After Photo bleach (FRAP) and Photo aAtivation (PA) provides a tool for precisely controlled and targeted appli-
cation of the selected laser beam to initiate photo-effects, which include (FRAP) fluorescence recovery, (FLIP) rate of loss with repeated
bleaching, (PA) activation and even ablation (with Q-switched UV laser). Imaging the effects reveals qualitative and quantitative informa-
tion about the cellular structures of interest and their environment. For example, photo-acti-
vated fluorescent proteins are becoming widely used to provide a spot light on small num-
bers of activated molecules, while UV uncaging is used to deliver agonists to specific cell
compartments and photoactivation is used in drug discovery to study detailed effects of
photo-sensitive topical compounds.

FRAPPA techniques allow measurement of molecular recruitment rates, trafficking and
turnover in single cells and sub-cellular organelles and applications include membrane and
protein binding, mytosis and cytoskeleton function. In these studies GFP is commonly used
in a fusion protein complex to label a protein of interest. The bleach or activation phase
“marks” those molecules for observation. Quantification of motion or recovery can provide a
useful measure of molecular mobility.
                                                                                                                              Typical profile of fluorescence intensity collected during a FRAP
FRAP is used increasingly in analytical devices to determine the identity of unknown sub-                                     experiment. For more information see the following reference:
                                                                                                                              Sprague, B., R. Pego, et al. Analysis of Binding Reactions by
stances (based on diffusion analysis), understanding cellular transduction, and identifying                                   Fluorescence Recovery after Photobleaching. Biophys. J. 2004
ligand binding sites.                                                                                                         Jun;86(6):3473-3495




               Composite triple color image of a microtubule protein (EB1-GFP) imaged               Embryo Cell Division using the Andor Revolution XD system.
               with objective-type TIRFM (60x 1.45NA), incorporating the iXon DV887                 Courtesy of Paula Sampaio, University of Porto Rua Campo Alegre,
               back-illuminated running at 0.5 frames/sec. The different colors reveal              Portugal
               the dynamics of the microtubules over time: frame 1 = red; frame 10 =
               green; frame20 = blue. Courtesy of Dr Derek Toomre, CINEMA laboratory,
               Dept. Cell Biology, Yale University School of Medicine, CT.




Head Office                       North America                         Japan                                China
7 Millennium Way                  425 Sullivan Avenue                   7F Ichibancho Central Building       Room 1116 Zhejiang Building
Springvale Business Park          Suite 3                               22-1 Ichiban-cho                     No. 26 An Zhen Xi Li
Belfast BT12 7AL                  South Windsor, CT 06074               Chiyoda-ku Tokyo 102-0082            Section 3 Chaoyang District
Northern Ireland                  USA                                   Japan                                Beijing 100029 China

Tel: +44 (0)28 9023 7126          Tel: (860) 290-9211                   Tel: 81-3-3511 0659                  Tel: 86-10-5129 4977
Fax: +44 (0)28 9031 0792          Fax: (860) 290-9566                   Fax: 81-3-3511 0662                  Fax: 86-10-6445 5401




                                                                                                                                                                                     April 2008




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