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:

  Position	Mag	NA	Immersion	Working 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:

  Position	Label	Range (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):

  Name	Excitation (nm	Mirror (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.