Biological High-end System Imaging

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Analytical Technologies Molecular Devices High-End System Biological Imaging 아스크 / ASK 서울특별시 성동구 행당동 4-31 우편번호:133-070 Tel : (02) 466 2545 Fax : (02) 466-2546 http://www.askus.co.kr e-mail : [email protected] www.askus.co.kr MM ICS (Integrated Confocal System) Real time confocal System Analytical Technologies Molecular Devices The lower image identifies the region that was photo-bleached using 488nm excitation for duration of 250ms The MM ICS array scanner heralds a new generation of high speed multi-point confocal systems targeted at the study of dynamic processes in living cells. This latest innovation in confocal microscopy from VisiTech International provides superior benefits to other confocal microscopy technologies. The graph plots the time history of the average fluorescence intensity in each of the regions identified in the upper image The MM ICS provides faster scan rates without the frame to frame variations inherent in other multi-point scanning techniques and its variable scan rates enable simple synchronisation to eliminate image ‘streaking’. The MM ICS also preserves the exceptionally low photo-bleaching performance associated with multi-point confocal systems. Arrows on traces 1,2,3 and 7 highlight increasing delays in onset of fluorescence loss due to increasing distances from the photo-bleach site Traces 5 and 6 show recovery of fluorescence intensity following the photo-bleach. Trace 8 indicates an isolated structure 2D-FRAP Extension for MM ICS Real Time Confocal Fully integrated FRAP module. The 2D-FRAP module is totally contained within the MM ICS head and thus requires no additional valuable bench space Advantages • Fully integrated with MetaMorph for image acquisition and analysis • Real-time confocal imaging • Low photo-bleaching for long duration live-cell experiments • Selectable pinhole sizes for optimal imaging conditions • Fully self aligning system; Easy on-site filter exchange • Perfect synchronization between scanner, camera and other devices • No random striping or field to field intensity variations • Complete control and seamless integration of illumination intensity and wavelength selection Real-time 3D visualisation Real-time confocal imaging at speeds of up to 1000 fps. This coupled with the 3D-RTTM module supports on-demand ‘3D’ reconstruction of experimental data as it is being acquired. Variable Speed Synchronised Imaging The basic optical path consists of a stationary micro-lens array illuminated with an expanded laser beam. A galvanometer mirror scans the array over the sample and de-scans the returning fluorescence light. This light is separated from the illuminating beam by a dichroic mirror, and passes through a stationary pinhole array to create the confocal image. Optimised Scanning Performance Multi-Wavelength Compatibility (UV to near-IR) Selectable pinhole sizes permit users to vary the degree of confocality and throughput to match their experiment requirements for optimal imaging conditions. To simplify multi-labelled experiments, the compact mechanical design incorporates motorised dichroic and emission (barrier) filter changers. The appropriate choice of pinhole size for the stationary pinhole array enables full optimisation of experiment requirements in terms of objective magnification, spatial resolution and light throughput. Multiple laser line sources, either multiple-line lasers or multiple lasers in any combination, may be coupled with an AOTF for high speed (~microsecond) laser line selection and intensity control. Coupling of the excitation illumination is available by optical fibre or by direct coupling. Interface Device control MetaMorph applications Pinholes (software selectable) Scan rates Excitation changer Dichroic changer Emission (barrier) changer Dichroic alignment Camera synchronization Camera and microscope Microscope compatibility Wavelength range Integrated MetaMorph GUI MetaMorph device integration FRAP, Time lapse, Streaming, Multidimensional Analysis From 10μm to 64 μm diameter Up to 1000 fps AOTF (4 or 8 channels) Motorized, 4 positions, user exchangeable Motorized, 5 positions, user exchangeable Not required (self aligning) Included C-mount Upright and Inverted Near UV to Near IR 2 High-End System for Biological Imaging High-End System for Biological Imaging 1 Specifications www.askus.co.kr MM OptiGrid High Resolution in Confocal Quality Image without Expensive and Complicated Laser Systems Analytical Technologies Molecular Devices The MM OptiGrid slider is inserted into the field diaphragm slot of a standard illuminator on upright or inverted microscopes. This enables it to be used with any of the wavelengths available from the illumination system from UV to IR and also to be easily removed for standard microscope functions. Furthermore, it provides users with excellent imaging throughput and programmable experiment repeatability for parallel multiple-specimen imaging. Many modern microscope technologies are based on fluorescence. The challenge in fluorescence microscopy is to resolve even weakly fluorescent molecules in subcellular space. This requires microscopic systems with highest resolution and sensitivity as well as new methods. The structured light illumination of the MM OptiGrid method allows to get this extreme high resolution in confocal quality without expensive and complicated laser systems. It`s a new tool which is available for easy and reliable creation of optical slices in 2D and 3D. The MM OptiGrid system is now fully controlled using MetaMorph software, which ensures simple system integration. Utilizing the light emitted from an existing stabilized illumination source, the MM OptiGrid uses a two-dimensional optical grid mounted on a piezoelectronically driven actuator to project a line pattern onto the specimen. The grid is then moved perpendicularly to the grid lines, in 1/3 steps of its period length so that three grid movements result in one optical section. The MM OptiGrid is an excellent and cost efficient system for expanding the imaging capabilities of new or existing microscopes without taking up valuable laboratory space. Offering confocal-like image quality, the MM OptiGrid integrates quickly and easily into many microscopes. System Configuration • Digital Cooled CCD Camera • Stabilized light source : CoolLED • Z-Stage / Piezo focus system • Support for Olympus, Nikon, Leica, Zeiss microscopes The structured illumination process returns a strong signal wherever focus is sharp and aweak signal where focus is soft. Furthermore, a patented algorithm is used to remove the blur(weak signals) and to combine the strong signals in the grid image to optical sections comprising just the focal plane. A series of optical sections taken through a sample (z-stack) can then be combined to create a haze-free ultrasharp composite image. Image stacks can also be used to produce 3D reconstructions using our post processing software. 4 High-End System for Biological Imaging High-End System for Biological Imaging 3 www.askus.co.kr MetaMorph System Configuration MM 5D Imaging System : Timelapse, Multiwavelengths, Z Focus High quality, cost effective and fully support imaging systems By installing an externally attached filter wheel and Z focus control motor to a manual microscope, you will be able to select the wavelength and control the focal plane from MetaMorph. You can acquire data g p p q while switching between excited light and absorbed light, or while changing the samples focus. MM 3D Imaging System : Timelapse You can also select how to acquire data whether to simply store the data to memory or directly save it to the hard disk. With the abilities of MetaMorph Timelapse (Acquire Timelapse), you can simply alter the time interval during observation, and you can also pause, resume and stop observation. Shutter Shutter Filter Wheel Z focus Motor MM 4D Imaging System : Timelapse, Multiwavelengths With manual microscopes, you can control externally attached filter wheels with multi-filters using MetaMorph, allowing you to observe samples and switch in a moment between excited light and absorbed light. For electric powered microscopes, you can observe samples by automatically controlling the microscope’s filter cube turret without the use of an externally attached filter wheel. Filter Wheel MM 6D Imaging System : Timelapse, Multiwavelengths, Z Focus, Multi Position You can perform timelapse at multiple points by controlling the microscope’s focusing handle and electric powered sampling stage from MetaMorph, while simultaneously acquiring Z axis data from each of those points. Shutter Z focus Motor Timelapse Position 1 MM 4D Imaging System : Timelapse, Z focus Shutter control the Z axis with MetaMorph by equipping a Z focus control motor, for that microscope, to the manual microscope focusing handle. You can also acquire data with MetaMorph by setting the number of slices and slice intervals in relation to the samples thickness. Z focus Motor Motorized Stage Shutter Position 2 6 Z focus Position 3 High-End System for Biological Imaging High-End System for Biological Imaging 5 www.askus.co.kr MetaMorph High quality, cost effective and fully support imaging systems Acquire Scan Slide Multi Dimensional Acquisition (MDA) Users have the flexibility to set exposure time, binning mode, gain levels, camera speed, bit depth, sub region acquisition area, and other advanced cameraspecific parameters. Simultaneous acquisition from t wo cameras is supported. Scan an area larger than the field of view by acquiring multiple images and stitching them together-ideal for large tissue samples. Microscope and hardware controls and illumination, magnification, and XYZ location settings are easily accessed from the menu. MetaMorph’s customizable auto -focus capabilities keep lengthy time dependent events in focus Integrated Morphometry Analysis (IMA) Region Measurement Measure original area (total, whole, pixel, etc.) calculations, perimeter, center points, brightness center position, width/ length, shape factor, average intensity value, OD, etc. Measure ROI(Region of Interest) area, intensity etc for single image or stack image. Graph Intensityes Filters & Stack Arithmetic Measure original area (total, whole, pixel, etc.) calculations, perimeter, center points, brightness center position, width/ length, shape factor, average intensity value, OD, etc. Make Movie Montage 3D Reconstruction Create movie (AVI, Quick time format) Display image from Stack file Create a reconstructed image by designating the rotational direction and rotational angle regardless of whether the bright field or fluorescent image. Multi Dimensional Motion Analysis Track 3D object and 2D object automatically. Easy to define and result. Overlay Image 2 overlapping specimens different color pigment Overlay 7 fluorescence images include DIC. And correcting position Auto Align Track 3D object and 2D object automatically. Easy to define and result. Multi-Viewer (Review Multi-Dimensional Data) Acquired multidimensional image data. Best Focus feature, you can extract frames that are automatically focused from 3D images and run finely focused timelapse movies 8 High-End System for Biological Imaging High-End System for Biological Imaging 7 Colocalization Review Multidimensional Data (RMD) www.askus.co.kr MetaMorph Application Modules High quality, cost effective and fully support imaging systems Angiogenesis Tube Formation Application Module Cell Health Application Module • Analysis of up to three fluorescent probes for cell-based apoptosis and necrosis assays • Classification and quantification of viable, early and late apoptotic cells, necrotic cells • Better quantitation by creating a single in-focus composite image from multiple Z-series images • Multi-parameter analysis measurements include tube length, number of branch points, number of nodes and more Adherent Chinese Hamster Ovary (CHO-K1) cells were incubated with various concentrations of staurosporinefor 6-12 hours. Top left: control, top right: 0.1 μM staurosporine, bottom left: 3 μM staurosporine, bottom right: measured (Green: viable, blue: early apoptotic, purple: late apoptotic, red: necrotic). Left: a best focus image is obtained from a Z series, right: The application module identifies tubes (white) and nodes (green). Cell Cycle Application Module Granularity Application Module • Classification and quantification of cells in different stages of the cell cycle • Option to use specific apoptosis and mitoticsstains, for increased classification accuracy • Count punctuate objects • Designed for receptor internalization or clustering target molecules • Choice of six granularity indices • Measurements include count, total and mean area, and intensities of granules and nuclei U2OS cells, top: overlay, bottom: the module identifies granules and nuclei, even in cells with high background (red arrow). Left: CHO-K1 cells stained with Hoechst 33342. Right: The Cell Cycle module identifies cell cycle phases: G0/G1 (dark blue), S (light blue), G2 (green), Early M (orange) and Late M (red). Count Nuclei Application Module Live/Dead Application Module • Automatically counts nuclei and captures intensity measurements • Accurate segmentation of touching cells • Classification and quantification of live and dead cells • Measurements include counts and percentages of live and dead cells, per-wavelength measurements and more CHO-K1 cells incubated with DMSO for 6-12 hours prior to staining. Cells were labeled with H33342 and PI diluted in PBS for 30-60 minutes before image acquisition. Top left: control, top right: 20% DMSO, bottom: measured (red: dead, green: live). Cell Scoring Application Module • Identification of two sub-population of cells • Ideal for counting and logging measurements of cells in twowavelength experiments The module can be used as a segmentation tool to identify two different stains or to score cells. • Classification and quantification of mitotic and interphasecells • Measurements include count, percentage and wavelength-specific intensities of mitotic and non-mitotic nuclei, and more Multi Wavelength Cell Scoring Application Module • Multi-parametric analysis of up to seven wavelengths • Identification of sub-populations of cells • Measurements include scoring profiles, wavelength-specific count and percentage of negative and positive cells, cell-by-cell wavelengthspecific stained area, integrated and average intensities Top: two wavelength images, bottom left: overlay, bottom right: segmentation. • Designed for the measurement and analysis of neuriteoutgrowths • Works with or without nuclear stain • Measurements include total neuriteoutgrowth, total branches and cell bodies, straightness and more Left: CHO-K1 cells treated with Nocadazolefor 18 hours before staining with anti-phospho-HistoneH3 (Ser28). Right: mitotic (green) and interphase(red) cells. 10 Monopole Detection Application Module • Classification and quantification of mitotic cells with monopolaror bipolar spindles • Measurements include count and percentage of monopoles, bipolesand interphasecells, DNA structures area, cell classification and more 3T3-L1 mouse fibroblast cells treated with monastroland stained with mouse anti-beta tubulinprimary antibody detected with a FITC conjugated goat anti-mouse secondary antibody. Nuclei are stained with Hoeschst33342. Left: monastrol, right: segmented image shows interphasecells (red), bipolar spindles (blue) and monopole (green). High-End System for Biological Imaging High-End System for Biological Imaging 9 Neurite Outgrowth Application Module Mitotic Index Application Module www.askus.co.kr MetaMorph System High quality, cost effective and fully support imaging systems Analytical Technologies Molecular Devices MOSAIC System LIVE CELL FUNCTIONAL IMAGING 4D Viewer/ 3D Measurement Option Visualization of multidimensional data sets, stacks and sequential images. Users can simultaneously view multiple z sections, wavelengths, time points and stage positions in a single intuitive viewing window, as well as binarize multidimensional image data into discrete objects, for 3D isosurface viewing and rotation. Fluorescence microscopy is a powerful tool for studying inter-and intramolecular interaction of biomolecules in living cells. Imaging techniques such as photoactivation, fluorescence recovery after photobleaching (FRAP), fluorescence loss in photobleaching (FLIP) and fluorescence resonance energy transfer (FRET) are very sensitive and precise methods that allow the interrogation of cellular and molecular function in living cells. DMD technology in the MOSAIC system The Digital Micromirror Device technology in the MOSAIC system uses an array of micromirrorsto create custom illumination masks of any size or shape. Each micromirroris individually controlled and light is directed to the specimen only by activated mirrors that correspond to the illumination mask. FRET System (Fluorescence Resonance Energy Transfer) FRET involves the non-radiative transfer of energy from a fluorophore in an excited state to a nearby acceptor fluorophore. FRET may occur when fluorophores are within angstroms of one another. This technique is used to infer protein-protein interaction andcolocalization. A UNIQUE DIGITAL ILLUMINATION SYSTEM The system precisely controls the spatial pattern of illumination using Digital Micromirror Device (DMD) technology. A DMD consists of an array of micromirrors whereby each micromirror is a precise switch that enables light to be digitally controlled.A mask of any size or shape, including multiple discontinuous regions, is created in MetaMorph. The mask is applied to the DMD array and the specimen is illuminated only by the light directed by the activated micromirrors.light hitting the remaining unmasked regions is deflected out of the optical path. Targeted illumination masks using MetaMorph software Custom illumination masks are easily created in MetaMorph using standard drawing tools (e.g., line, circle, polygon, free-hand, etc.). Masks can also be created by segmentation overlays or automatically using MetaMorph software’s advanced drawing and data processing tools. MONITOR PROTEIN DYNAMICS OVER TIME MetaMorph provides powerful tools to conduct timelapse experiments using the MOSAIC system for ease of targeting and monitoring protein dynamics over time. MetaMorph has an automated calibration routine to ensure precision targeting, ability to define multiple points at which to display the mask during a timelapse experiment, and configurable time intervals. FRAP System (Recovery After Photobleaching) FRAP is a live-cell imaging technique used to study the mobility of fluorescent molecules. Both MetaMorph and MetaFluor provide mechanisms for triggering photobleaching, background subtraction, and shading correction. Easily monitor protein dynamics over time FRAP of GFP-actinfilaments in living cells. A: Baseline image before photobleaching. B: Overlay image showing targeted illumination regions. C: Simultaneous photobleachingof all regions with 488 nm laser. D: Image of fluorescence recovery 10 seconds after FRAP. Courtesy of Photonic Instruments and Dr. Clare Waterman-Storer, Woods Hole Physiology Course. Most importantly, both support high speed acquisition from low light level cooled CCD cameras. MetaFluor specifically provides real time measurements and graphing of DeltaF/F ratios from multiple regions of interest as the experiment progresses. 11 12 High-End System for Biological Imaging High-End System for Biological Imaging MicropointLaser System The MicroPoint laser is a fiber-optically pumped dye pulse laser system designed for beam alignment to one area of a target. Live cell studies, such as Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Loss In Photobleaching (FLIP), demand the rapid acquisition and lowlight level imaging of highly-sensitive, cooled CCD cameras with high quantum efficiency, low noise and fast readout rates. sample application: FRAP Top: pre-bleach, bottom: post-bleach. MetaMorph Slide Loader System Analytical Technologies Molecular Devices MetaMorph Slide Loader System The PL-200 is now fully controlled using MetaMorph® software via a USB connection, which ensures simple system integration. The award winning Prior Scientific PL-200 is an innovative high speed automated slide loading system that is ideal for a variety of high throughput microscopy applications. Higher Speed TIRF Microscope System Total Internal Reflection Fluorescence (TIRF) can provide the field of Life Sciences with new knowledge about the molecular mechanisms at the boundary to the cell interior. This is a highly complex application, as even the quickest membrane processes with very weak signals have to be made visible using extremely short exposure times. The demands on the technology are correspondingly high. To date, TIRF has been the only method that could be used to examine processes in the cell membrane environment and, until today, it has been associated with complex setups, time-consuming handling and high investment costs. Laser TIRF system. Slide throughput is dependent on the loader and the automatic scanning stage. Cycletimes are dependent on several factors and typically fall in the range 20-25s but can be configured for cycle times as fast as 15s. Compact Design Takes up a minimum of bench space thanks to the rotating arm design. Thisapproach also minimisesvibration and noise which means the system can quietly go about its business of processing slides. Stability is ensured by the presence of a rigid, common baseplatefor both the slide loader and the microscope. Easy to Use The simplicity of the slide transfer mechanism provides efficient, reliable slide loading. A single arm functions to pull the slide from the cassette to the stage and then return it with an opposite pushing motion. Once the slide is transferred to the stage it is located with a unique fixturingdevice for repeatable positioning and slide reloading in all axes. As the precision XY stage scans the slide, the transfer mechanism is completely clear of any interference with normal microscope function. 14 Flexible & Simple Flexibility is guaranteed by the random access capability. Cassettes are configurable as input only, output only or input/output depending upon your specific requirements. This makes the system ideally suited for high volume screening, analysis, classification and review applications Epi-Fluorescence TIRF TIRF Automated micrometer High-End System for Biological Imaging High-End System for Biological Imaging 13 www.askus.co.kr MetaFluor Fluorescence Ratio Imaging The MetaFluor software is the ratio imaging private software in order to do the ion concentration measurement and the like inside the cell making use of the indicator of fura2, BCECF, Indo1 and Fluo3 etc. Fluorescence ratio imaging is the monitoring of live cells in which a fluorescent indicator of intracellular ions is introduced. Indicator dyes have been designed to shift their fluorescence excitation or emission spectrum when binding with specific ions. Images are obtained at two different wavelengths, typically matching the absorption bands at the high and low binding conditions. The system provides simultaneous display of the raw data, ratio image, graphs of intensities, ratios and ion concentrations, and a non-ratiometric image such as a brightfield or phase-contrast image. Two different ratiometric indicators can be imaged and measured simultaneously. Save Data • Save hard disk and external media from image, event mark, data log, measuring range, calibration, all protocol • Save TIFF file format compressed ratio with LZW. • Make movie file (AVI) Measurement • Select of measurement object area rectangle ellipse with free rectangle, hand limitlessly • Average strength inside (area, ratio and calibration) density measurable • Export all measurements to either a text file or to a spreadsheet program such as Microsoft® Excel Calibration • Standard Grynkiewicz equation, Titration calibrations function, the system it proofreads pH and Ca density direct indicatory possibility Automation of Experiment Acquire • High sensitivity cooling digital CCD camera, analog video camera • Independent in every corresponding wave length and camera gain and exposure time setting possible • Select binning mode and ROI • Real time background shading correction • Real time ratio measurement • Z axial scanning measurement (separate axial control driver is needed) • Streaming data acquisition (DG-4/piezo Z axial drive) • Use journal function customization of the measurement condition • One button processing task bar register • Define Sequential journal • Make specific journal using TTL trigger (easy to use external equipment) Support pCLAMP 10 Software pCLAMP 10 software get data from Molecular Devices corporation (Axon Instruments)’s Digidata 1340A, 1440A. and it is possible to support MetaFluor. Time-laps • Interval time setting possible (streaming mode : high speed acquisition) • For background removal of measurement image in wavelength threshold setting possibility • Monochrome, pseudo color display using custom LUT • Monochrome, pseudo color display ratio image using IMD ( IMD : Intensity Modulated Display) 15 16 High-End System for Biological Imaging Ratio Imaging System based on MetaFluor High-End System for Biological Imaging MetaVue precisExcite Cost-Effective Integrated System for Bioimaging The MetaVue™ Imaging System is the cost-effective solution for basic imaging applications such as digital photography, multi-wavelength fluorescence, image processing and image analysis. MetaVue is a simple, easy-to-use system for acquiring and processing images, performing graphics functions, and archiving and retrieving images. FLUORESCENCE EXCITATION SYSTEM CoolLED uses the latest LED technology for its family of excitation products. From a simple single-wavelength source to multiple wavelength automated systems, all CoolLED products use active-cooling to ensure the best possible performance. As a result, stability, intensity and precise control of the LED excitation is available. There is an increasing range of LED wavelengths to choose from. Acquisition For multi-mode microscopy, MetaVue is able to acquire multiple fluorescence wavelengths combined with a transmitted light image. Images can be stacked into one file and saved as a Stack file format, then processed or measured all at once in one easy step. Key Features AutoQuant X Advanced Deconvolution processing software AutoDeblur® offers the most powerful deconvolution tools availablefor life science research.Retrieve better data from your images using the most complete suite of 2D and 3D algorithms available. It offers an advancedBlind Deconvolution algorithm which is both iterative and constrained. 17 18 High-End System for Biological Imaging All microscopes are limited by the laws of physics, and these laws state that when light passes through a medium, that light will bend. This is one of the most common causes of haze and blur in microscopy images. Deconvolution can correct this problem, not only removing the haze and blur, but restoring vital detail to datasets. • Modular system which drives up to 4 wavelengths • Switch on and off instantly • Adjust light intensity in 1% steps (0-100%) • Stable, repeatable light intensity • LEDs last for tens of thousands of hours • Wide range of LED wavelengthsavailable • Integrated with many imaging softwarepackages • Quiet & cool operation • Accepts excitation filters for fast-switching applications • Never have to re-align your light source • No warm-up or cool-down period required • No need to use ND filters • No need to use shutters • No bulb safety and disposal issues High-End System for Biological Imaging MAC 6000 System Automation System Microscope Automation System XY Stage, Z Focus, Shutter Filter Wheel + Shutter Motorized XY Stage : Ludl BioPrecision2 Filter wheel module, to control maximum of 2 filter wheels, 3 shutter units. The filter wheel of Stepper type has prepared 25mmφ and 32mmφ as a filter size. 1 MAC5000 based system loaded 2 filter wheel module. Maximum of 4 filter wheels and 6 shutters can be controlled with 1 controllers. Optimum to multiple Dimension data acquisition system and FISH system which include multicolored. Very useful use exciter and emission for Fura2 and FRET system. Live-cell microscopy often requires the use of environmental chambers, probes, manipulators and large specimens. The BioPrecision2 flat-top stage addresses these requirements with a completely flat, open top surface. The stage is fully compatible with all of the standard LEP inverted stage inserts and accessories. In addition, a unique removable “mini breadboard” area is available on both the left and right sides of the stage to provide a convenient point for attachment of manipulators and probes. Position Feedback Option XY Stage Control XY electromotive stage by μSTEP stage module 2 is added to the MAC5000 based system. Prepare the stage upright type microscope, inverted type microscope. Optimum to Multi Dimension data acquisition FISH System screening data acquisition. Inverted Microscope Upright Microscope Resolution (minimum) Standard Open loop 50nm Rotary Encoder 200nm Linear Encoder 100nm High Res. Linear Encoder 50nm Standard Open loop 25nm Rotary Encoder 100nm Linear Encoder 100nm High Res. Linear Encoder 50nm MD XY Stage Max. Speed (mm/sec) Repeatability 0.75um 60 mm/sec 120 x 100mm 0.25um 2um 0.20um 0.75um 6um 0.60um 30 mm/sec Travel 6um 0.60um 30 mm/sec Accuracy 100 x 75mm 0.25um 2um 0.20um MD Z focus System Z Focus Z motor module is added to the MAC5000 based system. Corresponds to the various microscope. Piezo (Z Axial high-speed scan unit) Furthermore with addition, it is possible in the system which controls the XY stage of the MAC5000 based system to install Z axial highspeed scan unit. Piezo-electricity is installed in the stage holder part, it is possible to use with all object lens which have been installed in the microscope Travel Range 200 μm 500um Repeatability 1 nm 4nm Accuracy (full travel) 0.50% 1% Resolution 16bit 16bit Settling time (5 μm move 150g load) 25ms 37nm Maximum Load 500g 400g X axis ±55mm, Y axis ±37.5mm Resolution 0.36°(0.72°) Resolution 0.002mm Interface RS-232C Feed screw lead 1mm Motor Cable 1M Moving Guide V groove and the cross-roller Weight 1.5kg Repeatability ±0.002mm Maximum speed 16mm / s (8,000pps at) Memorize 3 position : Home, Top, Bottom Uniblitz Shutter Driver 20 The VMM-T1 is a complete timer/driver system for normally open or closed shutter operation. For precision control, flexibility, accuracy, and repeatability, it incorporates every feature necessary for most applications. The VMM-T1offers an exposure and delay interval range from .1ms to 2.8 hours. TheVMM-T1 also provides three choices for both internal timer activating and resetting. In addition, the shutter can be controlled from the BNC inputs, these inputs can also be controlled via a computer serial port(RS-232C). By selecting the proper address for each unit, up to 8devices can be controlled from one serial port. High-End System for Biological Imaging High-End System for Biological Imaging 19 Travel VCM-DI VMM-D3J VMM-TI www.askus.co.kr Illuminator Polychrome 5000 monochromator The Polychrome 5000 is a versatile and ultra-fast switching monochromator for fluorescence microscopy. It provides the flexibility to adjust the excitation wavelength in order to optimize an experiment even under non-standard or unknown conditions. It offers speed, stability and brightness to acquire the best possible data from living cells. Lambda DG4/ DG5 : Ultra High Speed Wavelength Switcher DG4 includes xenon lamp (175W or 300W) which enables fast light path switching (1.2 ms) through galvano mirrors. Unlike to mechanical filter wheels, DG4 perfectly eliminates mechanical vibration because the emitted light passes through optical fibers to the microscope. Also, acts as a darkening filter (100%, 50%, 33% transmittance), controlling the light source intensity by slightly retarding the axis of light path. Monochromator Scanner mounted diffraction grating Wavelength range 320 nm to 680 nm Scanning speed Up to 400 nm/ms Half-power bandwith 14 nm Motorized bandwidth (optional) 2 nm -15 nm, 1 nm resolution Motorized Intensity (optional) 0% -100%, 10% resolution Output 330~700nm (ozone free) Output power > 10 mW (@470 nm @ 14 nm BW, with new lamp) Light guide output 7.5mW@340nm, 8.2mW@380nm Optical fiber UV/Vis quartz/quartz fi ber, NA 0.22, length 2 m or 3m Lamp Type 175W or 300W xenon Light source 150 W Xenon high stability lamp Lamp life time 500 hours (with usage condition 1000 hours) Filter size diameter DG4 : 25mm, 4 mounted DG5 : 25mm, 2 mounted, 18mm, 3 mounted Shutter open, closing speed 500μsec Light guide length 2M Simultaneous-Imaging System : Multichannel Imaging: DV2, DC2, QV2 Lambda – LS : Stand-Alone Xenon Arc Lamp and Power Supply Independent type of xenon illuminant unit (175W or 300W) with, at the optical fiber possible ) , p use into the microscope. Possible to use excited filter of LB10 series. Robust application dislikes the vibration of the filter wheel. Output Standard bulb 340nm to IR Lamp Type 175W or 300W xenon Radiant Output 25 Watts (175W lamp) (broadband, full beam) Lamp life time 500 hours (with usage condition 1000 hours) Lambda 10 Series : High Speed Filter Wheel high performance, microprocessor-controlled multi-filter wheel system for imaging applications requiring up to 3 filter wheels Filter change time 40 msec between adjacent filters (10 position wheel) 31 msec between adjacent filters (3 position wheel) Control 3 wheels and up to 2 optional shutters Shutter SmartShutter™ and Uniblitz® shutter Applications Lambda SC : SmartShutter Controller Lambda SC is a full-featured microprocessor controlled shutter driver for the SmartShutter. It supports the same shutter command set used for our other Lambda controllers and offers enhanced performance through additional of modes of operation. The additional operating modes allow programmable control over the exposure time, and the delay between trigger and shutter opening. • Real-time multicolor imaging • FRET imaging • Fluorescence polarization/anisotropy imaging • Simultaneous fluorescence/DIC imaging • Drug discovery with Cy3/Cy5 • Single-molecule fluorescence (SMF) • pH imaging with SNARF • Multiwavelength TIRF • Multiwavelength FLIM • Voltage sensing with di-4-ANEPPS • Other emission ratio analyses • Fluorescence in situ hybridization (FISH) • cAMP imaging with FICRhR • Multichannel confocal microscopy when used in conjunction with a spinning-disk confocal • Calcium imaging with fluo-3/Fura Red™ (Molecular Probes) or dual-emission indo-1 imaging 22 High-End System for Biological Imaging High-End System for Biological Imaging 21 www.askus.co.kr Andor CCD Cameras Scientific CCD, ICCD, X-Ray & EMCCD Cameras iXonEM + 860 CCD Sensor CCD Format Digitization Well depth Clara CCD Sensor CCD Format Digitization Well depth Readout Rate Linearity Cooling(°C) Dark Current Frame Rate SONY ICX285 sensor; interline-transfer 1392 x 1040 pixel, 6.45 x 6.45 um pixel size, 8.98 x 6.71 mm 16 bit @ 1 MHz 14-bit @ 10 MHz and 20 MHz 18,000; (30,000 2 x 2 binned) 20, 10, 1 MHz 1% (Maximum) -45 fan on, -20 fan off 0.0004 e-/pixel/sec @ -45 °C 11.6fps (max.) Readout Rate Linearity EM Gain Cooling(°C) Dark Current Frame Rate Back Illuminated Transfer EMCCD, >90% QE 128 x 128 pixel, 24 x 24 um pixel size, 3.1 x 3.1 mm 14 bit @ 10, 5, 3, 1MHz readout rate (16bit available @ 1MHz) 160,000 e-, typical 220,000 e-, Maximum 880,000 e-, typical Gain Register 10, 5, 3, 1MHz 1% (Maximum) 1 – 1,000 times (software controlled) -85 (Air) 0.002 e-/pix/sec @ -85 °C 0.02 @ 1000x gain and -85 °C EMCCD-Amplified Background Events 500fps up to several thousands iXonEM + 897 LucaEM S CCD Sensor CCD Format Digitization Well depth Readout Rate Linearity EM Gain Cooling(°C) Dark Current Frame Rate CCD Sensor CCD Format Digitization Well depth Interline-transfer EMCCD, QE up to 52% 658 x 496 pixel, 10 x 10 um pixel size, 6.58 x 4.96 mm 14 bit 26,000 e-, typical 100,000 e-, typical Gain Register 13.5 MHz 1% (Maximum) 1 – x1,000 times (software controlled) -20 (ambient air at 20ºC) 0.05 e-/pix/sec @ -20 °C 37.2fps Readout Rate Linearity EM Gain Cooling(°C) Dark Current Frame Rate LucaEM R CCD Sensor CCD Format Digitization Well depth Readout Rate Linearity EM Gain Cooling(°C) Dark Current Frame Rate Transfer EMCCD, QE up to 65% 1004 x 1002 pixel, 8 x 8 um pixel size, 8 x 8 mm 14 bit 30,000 e-, typical 80,000 e-, typical Gain Register 13.5 MHz 1% (Maximum) 1 – 1,000 times (software controlled) -20 (ambient air at 20°C) 0.17 e-/pix/sec @ -20 °C 12.4fps Back Illuminated Transfer EMCCD, > 90% QE 512 x 512 pixel, 16 x 16 um pixel size, 8.2 x 8.2 mm 14 bit @ 10, 5, 3, 1MHz readout rate (16bit available @ 1MHz) 160,000 e-, typical 220,000 e-, Maximum 800,000 e-, typical Gain Register 10, 5, 3, 1MHz 1% (Maximum) 1 – 1,000 times (software controlled) -85 (Air) 0.001 e-/pix/sec @ -85 °C 0.005 @ 1000 x gain and -85 °C EMCCD-Amplified Background Events 35 – 549fps iXonEM + 888 CCD Sensor CCD Format Digitization Well depth Readout Rate Linearity EM Gain Cooling(°C) Dark Current Frame Rate Back Illuminated Transfer EMCCD, > 90% QE 1024 x 1024 pixel, 13 x 13 um pixel size, 13.3 x 13.3 mm 14 bit @ 10, 5, 3, 1MHz readout rate (16bit available @ 1MHz) 80,000 e-, typical 730,000 e-, typical Gain Register 10, 5, 3, 1MHz 1% (Maximum) 1 – 1,000 times (software controlled) -80 (Air) 0.001 e-/pix/sec @ -75 °C 0.005 @ 1000 x gain and -75 °C EMCCD-Amplified Background Events 8.9fps iXonEM + 885 CCD Sensor CCD Format Digitization Well depth Readout Rate Linearity EM Gain Cooling(°C) Dark Current Frame Rate Transfer EMCCD, QE up to 65% 1004 x 1002 pixel, 8 x 8 um pixel size, 8 x 8 mm 14 bit 30,000 e-, typical 80,000 e-, typical Gain Register 35, 27, 13 MHz 1% (Maximum) 1 – 1,000 times (software controlled) -70 (Air) 0.028 e-/pix/sec @ -70 °C 0.012 e-/pix/sec @ -85 °C 0.005 e-/pix/sec @ -95 °C 31.4fps 24 iKon-L CCD Sensor CCD Format Digitization Well depth Readout Rate Linearity Cooling(°C) Dark Current Frame Rate Back Illuminated Transfer EMCCD, > 90% QE 2048 x 2048 pixel, 13.5 x 13.5 um pixel size, 27.6 x 27.6 mm 16 bit 100,000 e-, typical 5, 3, 1, 0.05MHz 1% (Maximum) -70 (Air) 0.001 e-/pix/sec @ -70 °C 0.00013 e-/pix/sec @ -90°C 0.00008 e-/pix/sec @ -100 °C 0.92fps High-End System for Biological Imaging High-End System for Biological Imaging 23 www.askus.co.kr Microscope Leica MM AF IX81-ZDC The IX81 begins with a unique two-tiered, V-shaped optical design that provides tremendous versatility without frame modifications and bright images with minimal reflections. Frame includes a motorized 6-position nosepiece, light path selector, and high-resolution Z-axis motor with 0.01um step size. Features • Consistent, drift-free focus throughout • More reliable experiment results • Ability to photograph more cells • Avoidance of illumination by excitation light • Ability to keep the cell active • Optimum photographic conditions at all times ZDC focus drift compensation method based on glass surface Bioimaging techniques contribute to a growing number of scientific breakthroughs. The Leica MM AF Imaging System powered by MetaMorph®, plays a large role in this revolution. With its image acquisition, processing and analysis capabilities, and complete set of tools for automation, Leica MM AF opens the door fornew insights into cellular function. Leica DM6000 • Customized system for all types of life science research • Automated transmitted light axis, including Leica’s unique, fully automated DIC • Motorized fluorescence axis with 5 or 8-position filter cube turret • Motorized 7-position objective turret with improved coding • New: convenient Leica SmartTouch touchscreen for controlling the automated modules • New: optional, external Leica STP6000 SmartTouch Panel for intuitive, convenient remote control of the microscope Zeiss Axio Observer ECLIPS Ti-E Microscopes are critical tools for cutting-edge research in biology, medical and pharmaceutical sciences. To satisfy the demands of today’s high-end research, Nikon has developed the new Ti series of microscopes. MetaMorph diverse image acquisition and analysis methods such as multi-dimensional time-lapse imaging to acquire temporal, spatial and spectral information of fast, dynamic live cell processes. Intelligently designed automation and further expansion Of Ti ideal for applications such as confocal, FRET, High Content Analysis (HCS), and photobleaching/ photo activation to study interaction of fluorescence protein molecules in living cells and tissues. Highly differentiated fluorescence techniques – in research on living cells these are becoming the standard. Carl Zeiss is focusing its knowledge and innovation on further developing these techniques. So that they may be made accessable to new applications all the time. At the forefront of the development of inverted microscopes stands is the Axio Observer – with the most efficient, flexible and gentlest fluorescence, protecting your cells and leading to brilliant results. Recognizable increase in convenience: TFT and LCD Input and monitoring station at the same time – the touch screen TFT display for the motorized Axio Observer. Z1 stand is opening up a new dimension in automated operation. 26 Perfect Focus System (PFS) The PFS employs high-performance optical offset, making real-time correction in the desired Z-plane possible. The state of the PFS is prominently displayed on the front of the microscope. Moreover, when the PFS is not in use, the optical component of the PFS can be simply retracted from the optical path. Ergotube Docking station Aqua Stop II High-End System for Biological Imaging High-End System for Biological Imaging 25 www.askus.co.kr Incubator System Chamlide WP Live Cell Instrument Chamlide TC Incubator system for various types of chambers • Chamlide TC can accept various Chamlide chambers and commercial culture dishes using 5 types of adaptors. • Chamlide TC includes special glass covers for disposable culture wares to maintain humidity and provide excellent transmission for imaging. (Lab-Tek chambered coverglass and chambered slide, 35mm dish) • The controller uses a flow meter to control the mixed gas (5 ~10% CO2/ 95% Air) level (using less than 50 ml/min). • Chamlide TC is available with in/out ports for perfusion or injection of liquids or drugs without having to open the incubator cover. • It is possible to control the temperature and gas concentration, program temperature and gas concentration gradients, and record the temperature and gas concentration over time using CCP ver 3.7 software or MetaMorph. Specifications Temperature range Recommended 5% CO2 / 95% Air flow rate Incubator physical dimension (mm) Heating method Humidifier bottle volume for humidity Sensor Incubator material IHS-101 Ambient +3℃ ~ 45℃ 40~50 ml/ min Cover 164.9 (W) x 134 (D) x 11 (H) Main body 160.0 (W) x 130 (D) x 14 (H) Adaptor 110.0 (W) x 62 (D) x 7 (H) Cover built-in heater glass Main body thin layer heater Humidifier cartridge heater Lens warmer thin layer heater Maximum 100 ml Thermo-couple Black anodized aluminum alloy Fluidic inline heater • Best for high throughput screening (HTS) while live cell imaging is proceeding. • Chamlide WP can be used on almost any microscope stage (depending on the stage type, the Chamlide WP requires a stage adaptor.). The Chamlide WP was designed to fit microscope stages with mounting frame K such as Zeiss or Leica standard stages or most XY-motorized stages. • The controller uses a flow meter to control the mixed gas (5 ~10% CO2/ 95% Air) level (using less than 50 ml/min). • Chamlide WP can accept various plastic-bottom or coverslip-bottom well plates (6, 12, 24, 48 or 96 well plates) as well as Chamlide PT (Well plate type magnetic chamber). • The offered special glass cover is available for disposable well plates to maintain humidity and provide good transmission for imaging Chamlide AC Water volume adjustable perfusion chamber Chamlide EC Electric stimulation chamber • In-line solution heating is the simplest and most effective way of warming perfusion solutions. • Minimal dead volume of 250 ㎕ • The body of the IHS-101 is connected to the supporting arm by a magnetic system for easy installation. Internal dead volume 250 ㎕ Inlet/outlet tubing size 0.8 mm (O.D) Heating method Thin layer heater Temperature range Ambient +3℃ ~ 45℃ Inline heater dimension 20 ø x 139 mm Supporting arm length 500 mm Main body acetal component Inline heater material Internal tubing steel stainless Supporting arm steel stainless, plastic or on/off agnet 28 Perfusion chamber 18ø coverslip type internal volume Maximum 0.6 ml Perfusion chamber 25ø coverslip type internal volume Maximum 1.5 ml Inlet/outlet port tubing size 1.0 mm (O.D) Chamber physical dimension Main body 55ø x 5T Bottom plate 55ø x 2T High-End System for Biological Imaging High-End System for Biological Imaging 27 Incubator system for commercial well plates & Chamlide PT www.askus.co.kr OsteoMeasure Bone histomorphometric analysis System Performing static and dynamic histomorphometric analyses of mouse bone specimen. Using the Osteomeasure analytic system, static parameters such as bone volume, trabecular number and cell counts as well as dynamic parameters of bone formation such as mineral apposition rates are quantified on non-decalcified bone sections. The core provides investigators with the data and will help them in evaluating their biological relevance. Extensive interactive Bone Histomorphometryanalysis system.Based on ASBMR Standard Nomenclature, uses specialized measurement list to automatically calculate over 175 parameters of bone. Intelligent open measurements may be redefined for experimental, unconventional or other uses. Automatically compensates for the kinetic labellingmethod employed. Sophisticated Report Generator integrates independent measurements from serial sections. Sophisticated Text File Generator allows composition of text files from primary or derived data that are compatible with all common spreadsheet-based packages -allows more sophisticated or unconventional analysis. Entire system menu driven and user friendly with extensive help system and bone specific prompts and options. OsteoMeasure parameter Specifications The following parameters specify the primary data which are available on a field by field basis and on a whole biopsy summary basis. The abbreviations used are in accordance with standard ASBMR nomenclature. (See J Bone Min Res Vol 2, No 6, 1987 595-610 for details.) Note that V1, V2, V3, S1, S2, S3, C1, C2, and C3 are used to denote user-defined measurements and can be renamed. T.Ar P.Ar(Lm) V1.Ar Tb.Pf.Ar O.Ar(Wo) V2.Ar B.Ar Ma.Ar V3.Ar Vd.Ar Md.Ar O.Ar Fb.Ar Tb.Pf.Pm O.Pm(Lm) Fb.Bd Rv.Pm L.Pm Al.Pm Mt.Pm V3.Bd B.Pm B.Bd(Lm) Ob.Pm Rm.Pm Al.B.Bd Mt.b.Bd V1.Pm S1.Pm Vd.Pm O.Pm(Wo) E.Pm dL.Pm Al.Md.Pm Mt.Md.Pm V2.Pm S2.Pm O.Pm B.Bd(Wo) Oc.Pm dL.Bd Al.E.Pm Mt.E.Pm V2.Bd S3.Pm B.Bd Md.Pm Q.Pm sL.Pm Al.Cm.Bd V3.Pm Widths Tb.Pf.Wi Fb.Wi v2.Wi Vd.Pf.wi W.Wi V3.Wi O.Wi Md.Wi O.Wi(Lm) Ir.L.Wi O.Wi(Wo) V1.Wi Numbers N.Sm N.Nc N.V2 N.Sm(Lm) N.Ot N.V3 N.Sm(Wo) NW N.C1 N.Ob N.Md N.C2 N.Oc N.dL N.C3 Areas Perimeters Additionally, the program derives the following static and dynamic indices of trabecular bone remodeling which are available on a whole biopsy summary basis. These derived quantities can be selected from combinations of up to five different primary data sources according to the user’s specifications. Windows Features This upgrade incorporates all the more popular features of the old system, but also many new features typical of Windows-based programs. Old users will notice, with relief, the use of features such as long filenames and arbitrary folder names for data as well as Windows-based printing and support for the Windows Clipboard for transporting data into other applications such as Microsoft Excel. Other Enhancements Volumes BV/TV OV(Lm)/BV OV(Wo)/TV V2.V/TV VD.V/TV OV(Lm)/OV OV(Wo)/BV V2.V/BV OV/BV Ma.V/TV V1.V/TV V3.V/BV OV(Lm)/TV Md.V/TV V1.V/TV Surface/Vol BS.V/TV BS/BV Vd.S/BS OS(Wo)/OS BI(Wo)/BI Oc.S/BS sL.S/BS V1.S/BS S1.S/BS Al.Md.S/BS Al.S/BS Mt.ES/BS OS/BS BI/BS Fb.I/BS QS/BS dL.S/BS V2.S/BS S2.S/BS Al.Md.S/Md.S Mt.BI/BS Mt.ES/ES OS(Lm)/BS BI(Lm)/BS Ob.S/BS Rv.S/BS sL.S/LS V2.I/BS S3.S/BS Al.ES/BS Mt.BI/BI Mt.Cm.I/BS OS(Lm)/OS BI(Lm)/BI Ob.S/OS(Lm) Rm.S/BS dL.S/LS V3.S/BS Al.BI/BS Al.ES/ES Mt.Md.S/BS Mt.S/BS OS(Wo)/BS BI(Wo)/BS ES/BS LS/BS dL.S/sL.S V3.I/BS Al.BI/BI Al.Cm.I/BS Mt.Md.S/Md.S Thicknessess Tb.Th O.Th(Lm) Ir.L.Th Vd.Th O.Th(Wo) V1.Th Tb.Pf.Th Fb.Th V2.Th Vd.Pf.Th W.Th V3.Th O.Th Md.Th Numbers N.Ob/T.Ar n.Oc/B.Pm N.Ob/B.Pm N.Oc/E.Pm N.Ob/O.Pm(Lm) N.Oc/Oc.Pm N.Ob/Ob.Pm N.Oc/T.Ar Structural Tb.Sp It.Pf.Th Tb.N It.th Tb.Pf.Sp Tb.Pf.N Kinetic MS/BA BFR/BV FP QP MS/OS BFR/TV FP(a+) Ac.F MAR Mlt Rs.P Aj.AR Omt Rv.P BFR/BS Tt.P Rm.P Surfaces Further enhancements include support of digital images and cameras, as well as accumulating data from multiple objective magnifications within a single data file. The system can now be set up to automatically recognize data from different preparation types (e.g. stains), and will automatically display all the data acquired within a single folder. High-End System for Biological Imaging 29 Simple Interface The interface is simply composed of 4 sections. The first is the familiar Windows menu bar for basic file management, calibration and magnification selection; the second is the familiar OsteoMeasurecontrol panel for navigating the specimen, selecting a few drawing tools and measurement selection; the third is the Display Tabs along the bottom of the menu which select different views of the data and give access to the configuration options; and the fourth is the Data window where measurements are made and different views of the data can be seen. The primary and derived quantities displayed above are not, in any way, intended to represent a closed or completed measurement system. As new parameters are quantified and their representations defined, they will be in cluded in updated versions. 30 High-End System for Biological Imaging OV/TV Fb.V/TV OV(Wo)/OV V3.V/TV