Confocal Microscopy Lab

• Inverted Stand DMI 6000
• Motorized Encoded Stage for Mark & Find and Tiling
• Advanced Optical Module VISIR with Scan Rotation
• Optical Outfit with EL6000 Illumination & DIC
• Super Galvo Z Stage for Live XZ Imaging and UltraFast Z Stacks
• Transmitted Light Detector for Brightfield Confocal
• Conventional Galvo Scanning System
• Resonant Scanning System for Live Imaging
• Five Channel Simultaneous Confocal Imaging
• Four PMT Detectors One Hybrid Detector with High QE
• Acousto Optical Beam Splitter (AOBS) Technology
• AOBS Provides Fully Automated Confocal Imaging with Ultra High Sensitivity
• RGB Lasers with Total 7 Lines: 458, 476, 488, 496, 514, 542 and 633nm
• UV 405 Laser with AOTF
• Premium Workstation
• Bread Board Top Air Table with Compressor
• Stage Top Incubation Tokai Hit System

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DMI6000 Fully Automated Inverted Microscope and Components:

• Fully Automated DMI 6000 Inverted Microscope "intelligent" Stand
  • 6 Position Fluorescence Axis Motorized
  • 6-fold Objective Nosepiece Revolver M25 thread
  • 8-fold Auto Fluorescence Intensity Module (FIM)
  • Tilting Ergonomic Binocular Tube
  • Scanner Port Left and Additional CCD Port Right
  • Illumination intensity and diaphragms set automatically to selected objective
  • Single keystroke contrast switching BF/POL/DIC/Ph/Fluo
  • 7 Freely programmable buttons to simplify User Specified Tasks
  • Motorized 5-fold coarse and fine focusing with sensitivity and parfocality memory
  • Control Box CTR 6500 with Interface for the transfer of microscope parameters to the SP5
• Baseplate DMI 6000
• Universal Mounting Frame
  • Petri Dishes with 24-68mm Diameter
  • Glass Slides with 24-120mm Length
  • Outer Dimension: 160x110 mm
• Z Galvo Insert for Petri Dishes, 36mm
• Motorized XY 3-Plate Stage
  • Mark & Find—multipositional timelapse, marking and returning to fields of interest, etc.
  • Tile scanning –automated juxtaposed field acquisition to create a "Montaged" superimage
  • Integral software control within LASAF
  • Mechanical control via STP6000 and/or Smart Move remote control devices field acquisition
• Optical Outfit DMI 6000 with DIC Components
  • Automated switch btw eyepieces, photo/video or TCS SP5 II port
  • Manual 3-Plate Mechanical XY Stage with holes for mounting Super Galvo Z Stage
  • Pair of adjustable eyepieces HC PLAN s 10x/25
  • Motorized Revolver for Objective DIC prisms
  • Motorized TL Condenser for Automated DIC
  • Condenser S28 with 28mm Working Distance and 0.55 NA
  • Filter system ICT, Analyzer and Polarizer for DIC
  • EL6000 External Fluorescence Light Source, 2000 hour bulb life, Alignment Free, Fiber Coupled to microscope for thermal stability
• Standard Cubes for Eyepiece Viewing
  • Long Pass DAPI Cube, UV-Excitation (BP 340-380, LP 425)
  • Long Pass GFP Cube, Blue-Excitation (BP 450-490, LP 515)
  • Long Pass RFP Cube, Green-Excitation (BP 515-560, LP 590)

Highest Grade Objectives for Confocal Imaging:

• Objective HCX PL FL 5X/0.15 Dry
  • FWD 12mm
  • Cover Glass (not critical)
• Objective HC PL FL 10x/0.30
  • Cover Glass 0.17 mm (#1.5)
  • FWD 11.00 mm Objective HC PL FL 20x/0.50
  • Cover Glass 0.17mm (#1.5) -FWD 1.15mm
  • DIC with K3 & D Prisms
• Objective HCX PL APO 40x/1.1 W
  • Cover Glass 0.17mm (#1.5)
  • FWD 0.63mm
  • High Transmission through 3601100nm
  • DIC with K7 and E Prisms
• Objective HCX PL APO CS 63X/1.400.60
  • Oil
  • FWD 0.1mm
  • Cover Glass 0.17mm (#1.5)
  • DIC with K10 and E Prisms

DIC Prisms for 20x, 40x and 63x Objectives:

• IC condenser prism K3
• ICT condenser prism K 7
• ICT condenser prism K 10
• IC Prism D IC Prism E

Super Z Galvo Stage

• Live XZ Plane Image Acquisitions
• Z-stack acquisitions at up to 15 frames per second at 512x512 image resolution on RS-Scanner
• Retains compatibility with standard DIC prisms unlike piezo focus devices
• 1500um travel
• 3nm step size
• 40nm reproducibility

Leica TCS SP5 II AOBS HyD TS System and Components

• AOBS Based Tunable Filter Free Spectral Confocal
• GaAsP Hybrid Detectors and PMTs
• Tandem Scanning with Conventional Galvo and Resonant Scanners
• Optical Module VISIR with Scan Rotation:
  • Optical coating VISIR with high transmission of > 99% throughout the visible and infrared spectrum ranging from 400-1300nm
  • Low reflection at optical surfaces throughout the visible and infrared spectrum
  • Independent, optical rotation of the scan field up to around 200° by a highly transparent Abbe–Koenig rotator design
  • Prepared to include either a standard or tandem scanner with Leica parallax-free patented three-mirror design
  • Prepared for use with 405 nm line laser, all line lasers with visible light, white light laser, infrared lasers and optical-parametric oscillator (OPO)
• AOBS Confocal Module
• TCS SP5 II Confocal Module with AOBS
  • Ultra High Sensitivity
  • Filter Free Spectral Detection with User Defined Bandwidth
  • Simultaneous use of up to 8 Lasers from UV to IR
  • Replaces the Conventional Rotating Dichroics with Fixed Optical Characteristics
  • Minimum User Supervision with Optimal Flexibility and Automated Design of the Optical Path & Bandwidth
  • Unmatched Flexibility: Mathematically equivalent to 40,320 Dichroics
  • Possibility of "All" combinations of the Excitation and Emission Wavelengths
  • Steep Transmission Cutoff, <2nm
  • Unmatched Spectral Integrity: Flat Spectral Behavior Results Undistorted Recording of Emission Spectra
  • Drastic Reduction in Phototoxicity with Low Intensity and Exquisite Signal Efficiency
  • Facilitates Better Time-Lapse of Living Cells
  • Low Maintenance with "No" Moving Mechanical Parts
  • Five Spectral Detectors with "Independently" adjustable Gain
  • Maximal Fluorescence Detection with Patented Prism Design
  • Automatic Adjustments of Pinhole Diameter
  • Manually Adjustable Pinhole for Improved Z Sectioning
  • Beam parking for FRET and FRAP
• 2100 Tandem Scanning System SP5 II Conventional Scanner:
  • Up to 2,800 lines-per-second
  • Up to 5 sustained frames-per-second at 512x512 image resolution
  • Up to 25 sustained frames-per-second at 512x16 image resolution
  • Beam-parking -Up to 8192x8192 image resolution
  • 2.12 mm Scanfield diameter with 10x
  • Rotation, panning, zooming capability
• Resonant Scanner:
  • Up to 16,000 lines-per-second
  • Up to 30 sustained frames-per-second at 512x512 image resolution
  • Up to 300 sustained frames-per-second at 512x16 image resolution
  • Up to 1024x1024 image resolution
  • 1.25 mm Scanfield diameter with 10x
  • Rotation, panning, zooming capability
• 5-Channel Leica "SP" Spectral Fluorescence Detection
  • Simultaneous detection of up to 5 fluorophores from 400nm to 800nm using prism mirror design for superior transparency (95%) and efficiency
  • Set emission bandwidth as small (specific) as 5nm and as large (efficient) as 300nm e.g. ECFP emission peak at 475nm480nm, broad-spectrum DAPI at 400nm-700nm
  • Set detection window wavelength minimum and maximum to the nanometer eg. FITC alone at 495nm-530nm or simultaneous FITC and TRITC at 495nm-514nm and 563nm611nm respectively.
  • Spectral imaging (ie. separation) of spectrally similar yet intensity dissimilar fluorophores through individual adjustment of amplification gain (voltage) control on MULTIPLE detectors eg. CFP-YFP, GFP-YFP
  • Separation of fluorophores without reference spectra in most cases (simultaneous - optical, sequential-optical, or channel-mathematical)
  • Compatible with fast-RS and TS-scanners for FAST spectral imaging!
• Transmitted Light Detector for DMI 6000
  • Auto Switching Mirror Confocal & Eyepiece
  • BF/DIC/POL/Phase Compatible (not for MP)
• Four Confocal Imaging Detectors
  • High Sensitivity Hamamatsu 9624
  • Actively Cooled for Extremely Low Noise
  • Low Dark Current
  • Extended Spectral Range with Increased QE
• Adaptor Kit for HyD SP Spectral Detectors

• Hybrid GaAsP Detector for Spectral Imaging
  • UltraSensitive Photon Detection with Maximum Quantum Efficiency
  • QE 2-3 times more than Standard PMTs
  • Broad dynamic range (16 bit)
  • Extremely low noise for increased S/N
  • True Photon Counting Capability
  • Improved Sensitivity Creates Very Low Photo-Toxicity and Improved Quality of Z Stacks
• Imaging Laser RGB for AOBS
  • R = "red" 10mW HeNe 633nm
  • G = "green" 1mW HeNe 543nm
  • B = "blue" 65mW multiline Argon (458, 476, 488, 496, 514nm)
• Laser Coupling for UV 405 nm Laser
  • Fiber Coupling, Optical and Mechanical Coupling Interface for UV laser
  • UV Adaptation Optics with Motorized 6-position Revolver
  • Chromatic Correction Device
  • Illumination Pinhole
  • Motorized turret for UV-Pinhole-Optics
  • Simultaneous use of all Lasers is possible
  • Switching between different types of microscopy stands is possible
• AOTF (Acousto Optical Tunable Filter) UV/405 nm
• Laser 405 nm, 50mW Laser System
• CorrL 405 20x/0.70 CS
  • Unique to SP5 II for UV Correction
• CorrL 405 40x/1.10 W CORR CS
  • Unique to SP5 II for UV Correction
• CorrL 405 40x/1.30 OIL CS
  • Unique to SP5 II for UV Correction
• CorrL 405 63x/1.400.60 oil CS
  • Unique to SP5 II for UV Correction

• Workstation PLUS with 2x19" LCDs
• Multitasking High Power Pentium Workstation with Windows 7 (64Bit) operating system
  • 12 GByte RAM
  • 160 GByte SATA hard disc drive
  • 1000 GByte SATA hard disc drive
  • Intel sixCore Xeon X5650 2.66 GHz
  • 2 TByte SATA Hard Disk Drive
  • 2x eSATA Interface
  • Dual Layer DVD Writer, DVD +/RW -10/100/1000 Ethernet Controller
  • Keyboard and Mouse
  • Two 19"LCD flat screens True color, 1280x1024 Pixels
  • MSWINDOWS 7
  • Leica LAS system software for control of scan process and image processing
  • Online software and hardware manual
• Computer Table
• SP5 Control Panel for Ergonomic Control of Multiple Confocal Functions
  • Panel Box with 6 Freely Programmable Digital Potentiometer Control Knobs
  • LCDs Display Current Assignment and Setting for Each Knob
  • Facilitates easy adjustment of scan parameters without taking eyes off image display
• LAS Live Data Mode
  • Recording of Manual and Automatic Workflows
  • Allows Correction of Background Fluorescence
  • Complex Time Lapse Measurements
  • Trigger Functions
  • Stack Profiling or ROI measurements
  • Bleaching Experiments and Online Display
• 3D Reconstruction & Animation
• LAS MicroLab Software Packages:
  • FRAP Wizard:
  • Step by Step Protocol
  • Efficient Bleaching by Automatic Zoom
  • Quantify the Measurement Data, On- and Offline
  • Export Data in *.xml format FRET Acceptor Photobleaching
  • Step by Step Protocol for FRET
  • FRET-efficiency will be calculated for user-defined regions and the overall result will be displayed as a FRET efficiency map
  • Export Data in *.xml format.
  • User Guidance with Background Correction and Cross-Talk Removal
  • On-and Offline Quantifying of FRET Efficiency
• SP5 Microscope Air Table Breadboard Top
  • 30" X 36" Stainless Steel Top, 4" Thick w/ M25 Holes
  • Custom Leg Set, 29" Height (matched to SP5 Table)
  • Casters for easy set up and positioning **Compressed Air Required Air Compressor for Table

• Tokai Hit GSI Chamber System for SP5 Super-Z Galvo Stage
  • GSI Stage Top Chamber
  • Objective heater
  • Heated Glass Lid
  • 3 Channel Temperature Controller
  • 1 Channel Gas Flow Controller (premix)
  • 35mm Dish Holder
• GSICGC Dish Attach for Chambered Coverglass