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

 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

Manual

A detailed description and manual: DRAGONFLY-01

More information: microscope feature description (ReasearchGate)

Contact: email