Andor Dragonfly

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General information

  Andor spinning-disc confocal (Dragonfly 200) based on an upright Leica microscope with automatic XY stage, motorised focus and neuroscience related live/physiological experiment support.

Application
  • Multi-channel confocal/widefield fluorescence imaging of fixed/live samples
  • Two-channel dual-camera simutaneously imaging
  • Multi-channel SRRF real-time super-resolution imaging with upto 110 nm lateral resolution
  • High speed burst imaging
  • Active illumination for selectve DNA breakdown, photo-bleaching and photo-activation imaging
  • Large free-stand XY stage for physiological devices
  • Selective pinhole size for high/low magnification objective
  • Dark chamber with Faraday shield
Accessibility
Microscope details
  • Microscope frame: Leica DM6 FS
  • Spinning-disc unit: Andor Dragonfly 200
    • switchable 40 um and 25 um (pinhole) spinning discs for high and low magnification objectives (typically 25x and 60x)
    • Switchable dual camera with optional 565 nm longpass and 565 shortpass dichroics
    • Andor 4-line ILE laser module
      • DAPI: 100 mW 405nm diode
      • GFP: 50mW 488nm diode
      • Cy3: 50mW 561nm OPSS
      • Cy5: 140mW 637nm diode
    • Quad-band emission filter for 405/488/561/640
    • separate emission filter wheel for each camera
  • Random access active illumination unit: Andor Mosaic 3
    • Mannually switchable light source
    • 1.1W 405nm laser
    • Port for 2nd laser
  • Objectives: 10x/0.32, 25x/0.95 W, 40x/0.8 W U-V-I, 63x/0.9 W U-V-I, 63x/1.4-0.60 Oil
  • Real-time super-resolution Imaging: Real time SRRF
  • Light source for sample Location: pE-300 white
  • Brightfield imaging mode: BF and DIC
  • Motorised XY Stage: Scientifica Motorised Movable Base Plate
  • Auto-focusing: N/A
  • Live experiment support: dark chamber with Faraday shield
  • Camera
  • Sample holder: options of slide and petridish
  • Optical bench: active
  • Commercial dervice contract: N/A
Filter settings
Confocal/widefield setup
Fluorophores DAPI GFP Cy3 Cy5 BF DIC
Laser 405 488 561 637 Halogen
Laser input dichroic Quad ET 405/488/561/637
Dichroic 565SP detected by IXon 888 EMCCD detected by Zyla sCMOS Any camera Any camera
Dichroic 565LP detected by Zyla sCMOS detected by IXon 888 EMCCD
Emission filters 450/50 nm (both camera) 525/50 nm (both camera) 600/50 nm (both camera) 700/75 nm (both camera) Empty

Sample inspection setup
Fluorophores DAPI GFP Cy3 Cy5 BF DIC
LED 377/50 485/20 560/25 640/13 Halogen
Filter cube Quad ET (405/488/561/640) BF/PH (Pos5) DIC (Pos6, Analyzer)

Condenser setup
Contrast mode Ph1 (Pos1) BF (pos2) IX-DIC20 (pos3) IX-DIC40 (pos4) IX-DIC60 (pos5)
Match objectives 10x ph1 all objectives 25x 40x 60x

Workstation and software
  • Acquisition software: Fusion
  • Active illumination software: IQ 3
  • Image processing software: IMARIS (linked with Fusion)
  • Image deconvolution: inbeded in Fusion
  • Acquisition workstation
    • Model: Dell Tower 7910
    • CPU: Intel Xeon E5-2637 V3 @3.5GHz (2 processors)
    • GPU: NVUDIA Quadro M4000, AMD Radeon ProWx 7100
    • OS: 64-bit Windows 10
    • RAM: 96GB
    • Local data drive (D:): 4 TB
    • Network name: cssd901402.ad.susx.ac.uk
Images taken from Dragonfly
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