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Nikon Spinning Disc Confocal

Nikon Spinning Disc Confocal

The Nikon Spinning Disc Confocal is an ideal microscope for long-term 3D live cell imaging. With seven laser lines and the external emission filter wheel, the SDC can observe almost any fluorophore that is used for live cell microscopy. Housed in a fully enclosed Oko Labs environmental chamber allows for steady controls to enable multi-day timelapse acquisitions. The equipped Andor 888 Ultra EMCCDs are extremely sensitive in order to greatly reduce both exposure powers and times. Using both cameras will allow the user to acquire two images simultaneously, which is ideal for real-time FRET imaging.

 Rules of Use:

  1. The maximum duration of a reservation from 9:00am – 5:00pm is 4 hours, except for on Mondays (please see #3). If you need more than 4 hours, like for an overnight experiment, please start your session after 4:00pm.
  2. If you run an overnight experiment, you are not allowed to reserve a session during peak hours (9:00am - 5:00pm) the following day.  Please end your overnight session by 9:00am the following morning.
  3. In general, please keep your Monday reservations to 4 hours. If you would like to do an experiment that lasts for more than 48 hours, you are allowed to start this on Friday afternoon at 4:00pm and continue until Tuesday morning at 9:00am.
  4. The microscope can be reserved for any amount of time starting at 4:00pm each afternoon and all day Saturday and Sunday (and Monday as outlined above).
  5. University holidays and cancellations (i.e. snow days) will be treated like a Saturday or a Sunday. 
  • Please click here for the Startup and Shutdown procedures for the microscope and the environmental control.
  • Lasers: 405, 445, 488, 514, 561, 594, 640
  • Objectives:
    PositionMagNAImmersionWorking Distance (µm)Characteristics
    1 10x 0.45 Air 4000 Plan Apo λ
    2 20x 0.75 Air 1000 Plan Apo λ
    3 40x 0.95 Air 250 - 160 Plan Apo λ DIC
    5 100x 1.45 Oil 130 Plan Apo λ
  • Emission Filters:
    PositionLabelRange (nm)
    1 CFP 482/35
    2 YFP 540/30
    3 mCherry 630/30
    4 TRITC 620/60
    5 GFP 525/50
    6 Quad 447/38
    520/22
    598/40
    705/68
  •  Camera: 2x Andor 888 Ultra EMCCD
  •  Key Specs:
    Active Pixels 1024 x 1024
    Pixel Size 13 x 13 µm
    Image Area pixel well depth 80,000 e-
    Maximum Readout Rate 30MHz
    Maximum Frame Rate 26 - 9,690 fps
    Read Noise < 1 e- with EM gain
    QE Max >90%
  • Spinning Disc Head: Yokogawa CSU-X1
    Scanning Method Microlens-enhanced Nipkow disk scanning
    Scanning Speed 1,500 - 5,000 rpm
    Port 1 Andor 888 Ultra EMCCD
    Port 2 Andor 888 Ultra EMCCD
    Filter Wheels Six positions each
  • Light Source for widefield: HBO arc lamp
  • Filter Cubes (for widefield):
    NameExcitation (nmMirror (nm)Emission (nm)Fluorophore
    DAPI 340 - 380 400LP 435 - 485 Dapi
    Hoescht
    Alexa405
    CFP
    Dual CFP/YFP 454 - 486
    518 - 559
    CFP
    YFP
    FITC 465 - 495 505LP 515 - 555 FITC
    GFP
  • Environmental Chamber: Oko Labs, CO2, Temperature, Humidity
  • Pixel Size on Image: (Click here for a nice overview)
    Objective
    Lens Magnification EMCCD Pixel Size 10x 20x 40x 100x
    1 13000 x 13000 1300 650 325 130
    1.5 13000 x 13000 866.67 433.33 216.67 86.67
  • Image pixel size = (Camera pixel size x bin factor)/(objective magnification x lens magnification x C mount magnification).
  • Pay attention to your units! The units in the table are nm.
  • Most often the lens magnification and C mount magnification = 1.
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