Andor Dragonfly 505

Location: room i04.U1.015 – book in PPMS

The Andor Dragonfly is an advanced imaging platform with multiple imaging modalities, Borealis Perfect Illumination, incubation chamber and CO2/O2 mixers.

Imaging modalities/features:

  1. Confocal (dual spinning disk with 25 & 40 µm pinholes)
  2. Laser-based widefield
  3. Total Internal Reflection Fluorescence (TIRF) Microscopy
  4. Super-resolution (SRRF)
  5. Astigmatic lens for 3D SMLM (single molecule localisation microscopy)

Specifications:

  • Nikon Ti2E inverted microscope (single deck)
  • 2 cameras: 1x Andor iXon 888 EMCCD and 1x Andor Zyla sCMOS
  • Spinning disk with two pinhole disk patterns: 25 µm and 40 µm in confocal mode
  • Camera zoom optics of 1.0x, 1.5x and 2.0x for optimum resolution across different objective magnifications and camera pixel sizes
  • Illumination zoom optics of 4x, 16x & 36x power density
  • Motorized stage:
    1. ASI XY stage with linear XY encoders: 50 nm resolution, 300 nm repeatability
    2. ASI Z piezo: Z position with sub-nanometre accuracy, 500 µm travel range
  • Incubation chamber (25-42 ºC)
  • Ibidi CO2/O2 gas mixer

Light sources:

  • CoolLED – pE-300 White Light source
  • 405 nm laser:  (100 mW) – Continuous-wave
  • 488 nm laser:  (150 mW) – Continuous-wave
  • 561 nm laser:  (150 mW) – Continuous-wave
  • 637 nm laser:  (140 mW) – Continuous wave

Filter for visual examination:

  • DAPI: Excitation: BP 377/50 Emission: BP 447/60
  • GFP/FITC: Excitation: BP 482/18 Emission: BP 520/28
  • DsRed/RHOD: Excitation: BP 531/40 Emission: BP 593/40

Objectives / position:

  1. CFI P-Apo 10x Lambda / NA 0.45 / WD 4.0 mm (MRD00105)
  2. CFI P-Apo 20X Lambda / NA 0.75 / WD 1.00 mm (MRD00205)
  3. CFI P-Apo 60x Lambda / NA 1.40 / WD 0.13 mm – oil (MRD01605)
  4. CFI Apochromat TIRF 60x / NA 1.49 / WD 0.13 mm – oil (MRD0169) – €€€
  5. Apochromat LWD λS 40x / 1.15 Water 0.60 mm (MRD77410) – €€€
  6. Open position (available for high-end rental objectives)
Extra high-end Objectives (Available via Automated rental System)
  1. CFI P-Apochromat 10x NA 0.50 /  WD 5.50 mm – Glycerol (MRD71120) – €€€ → rights required – info
  2. CFI P-Apochromat 20x NA 0.95 /  WD 0.95 mm – Water (MRD77200) – €€€ → rights required – info
  3. Other available objectives

Airy unit (AU) calculation for most objectives available for Nikon stand based spinning disks per pinhole size.

Airy unit (AU) calculation for the Nikon objectives available for the IOF spinning disks systems (for each pinhole size). Objectives highlighted in light blue are readily available for the Dragonfly system.
  • orange (AU < 0.9): high Z sectioning at the cost of compromised brightness/signal collection
  • green (0.9 < AU < 2): optimised Z sectioning & brightness
  • red (AU > 2): compromised Z sectioning

 Lightpath and cameras

image source

Cameras:
  1. Andor iXon Ultra 888 EMCCD Camera: 1024 x 1024 pixels; pixel size 13 x 13 μm2; image area 13.3 x 13.3 mm2
  2. Andor Zyla 4.2 Megapixel sCMOS camera: 2048 x 2048 pixels; pixels size 6.5 x 6.5 µm2; image area 13.3 x 13.3 mm2

Camera Emission Splitter (for 2nd Port camera)

  1. Dual camera switching mirror: use to switch emission light path between primary and secondary camera ports
  2. Dual camera dichroic 565 nm longpass: for dual colour GFP/RFP simultaneous imaging (shorter wavelength is reflected to vertical camera port)
  3. Dual camera dichroic 640 nm longpass:  for dual colour Cy3/Cy5 simultaneous imaging (shorter wavelength is reflected to vertical camera port)

Emission Filter wheel Camera 1 (Andor iXon)

  1. 445/46 bandpass filter: DAPI / BFP / Blueberry / Azurite (use with dichroics #1, 2 and 6 only)
  2. 521/38 bandpass: GFP, Venus, mNeonGreen, Alexa488, FITC, mWasabi. For use with dichroics # 1, 2 and 6 only
  3. 594/43 bandpass: RFP/TRITC, mRuby, tDTomato. (use with dichroics #1, 5 and 6)
  4. 620/60 bandpass: mCherry, Hc-Red1, mRuby
  5. 685/47 bandpass: Cy5, Cy5.5, iRFP 670/682, mCardinal (use with dichroic #6 only)
  6. Dual EM filter 488-561 – GFP/RFP
  7. 478/37 band pass: CFP, mCerulean, mTurquoise, DAPI. For use with dichroics #3, 4 and 5 only
  8. Penta EM filter 405-488-561- 640: (DAPI/GFP/RFP/Cy5)

Emission Filter wheel Camera 2 (Andor Zyla)

  1. 445/46 bandpass filter: DAPI / BFP / Blueberry / Azurite (use with dichroics #1, 2 and 6 only)
  2. 538/20 bandpass filter: YFP, Venus, mCitrine. (use with dichroic #5 only)
  3. 521/38 bandpass: GFP, Venus, mNeonGreen, Alexa488, FITC, mWasabi. For use with dichroics # 1, 2 and 6 only
  4. 594/43 bandpass: RFP/TRITC, mRuby, tDTomato. (use with dichroics #1, 5 and 6)
  5. 620/60 bandpass: mCherry, Hc-Red1, mRuby
  6. 685/47 bandpass: Cy5, Cy 5.5, iRFP 670/682, mCardinal (use with dichroic #6 only)
  7. Dual EM filter 488-561 – GFP/RFP
  8. Penta EM filter 405-488-561- 640: (DAPI/GFP/RFP/Cy5)

Computer (Fusion-Imaris workstation PC):

  • Intel dual quad-core Xeon CPUs
  • RAM: 96GB ECC
  • nVidia M4000 8GB
  • AMD W7100 8GB
  • 6x SSDs (4TB for images)
  • 10 Terabyte disk drive upgrade

Software: Fusion 2.2

System manual and additional material is available on SeaFile (login required).

More information: microscope feature description (ReasearchGate)