Nikon CSU-W1-02

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

 

System description and specification

The Nikon CSU W1-02 spinning disk microscope is an large Field Of View (17 x 16 mm) Spinning Disk microscope. The dual disk configuration provides flexibility in optimized image acquisition of samples varying sample size, for which the piezo stage provides fast and precise imaging of 3D stacks. The automated water immersion system allows prolonged and combined live imaging experiments. In the relatively near future, this system will be equipped with a micro-manipulator for a (1-sided) pipette aspiration setup.

Imaging modalities/features
  1. Widefield (LED-based): large FOV, using the Nikon camera
  2. CSU-based dual-disk Confocal (spinning disk 25 and 50 micron disks) – 2 sCMOS cameras (Teledyne Photometrics BSI)
  3. Widefield (laser-based): on 
  4. Feedback microscopy
Specifications
  • Nikon Ti2E inverted microscope
  • Yokogawa CSU-W1 dual-disk spinning disk unit with 25 μm & 50 μm pinholes disk
  • 3 cameras (1* CMOS + 2* sCMOS)
  • Motorized stage + piezo drive: i) Nikon TI2-S-SE-E XY stage with linear encoders: 100 nm XY resolution, 500 nm XY accuracy; ii) Mad City Labs Z piezo & linear stage encoder: travel range: 200 μm, 0.4nm min step size
  • Perfect Focus System (PSF)
  • Automated Water Immersion Dispensing System
Light sources
  • CoolLED pE-300 for epi-fluorescence imaging: 400nm, 460nm, 550nm
  •  Omicron LightHUB Ultra laser system for confocal imaging: 405nm, 488nm, 561nm, 640nm

Ti2 – Widefield

  • Nikon Ti2E, with automated water immersion
  • CoolLED pE300 White MB
  • Camera:  Nikon DS-Qi2 FX CMOS (large FOV widefield)
  • Filter cubes:
    1. DAPI-5060C – EX 377/50, DM 409, BA 447/60
    2. GFP-4050B – EX 466/40, DM 495, BA 525/50
    3. mCherry-C – EX 562/40, DM 593, BA 640/75
    4. Cy5-4040C – EX 628/40, DM 660, BA 692/40
Objectives / position
  1. CFI Plan Apo λ 10x air / NA 0.45 / WD 4 mm (MRD00105)
  2. CFI Plan Apo λ 20x air / 0.75 / 0.17-0.25 mm (MRD00205)
  3. CFI Apo LWD λ S 20x WI / NA 0.95 / WD 0.9-0.99 mm / Water (MRD77200) – €€€
  4. CFI Plan Apo λ 40x WI / 1.15 / 0.60 mm (MRD77410) – €€€
  5. CFI Plan Apo λ 60x oil / NA 1.4 / WD 0.13 mm / oil (MRD01605) – €€
  6. Free slot for ‘rental objectives’
 
Additional high-end bookable objectives (book in PPMS)
  1. CFI P-Apo 10X MultiCorr / NA 0.5 / WD 2.5 mm  (MRD71120) – €€€ – Oil, Water, Glycerin, Silicone Oil
  2. CFI Plan Apo 25XC Sil / NA 1.05 / WD 0.55 mm  (MRD73250) – €€€ – special suited for Live Imaging

CSU-W1 Confocal Scanner Unit

Light path and cameras

image source

Dichroic Mirror Block
  1. Confocal (visible lasers) quad-band dichroic mirror: 405 nm / 488 nm / 561 nm / 640 nm
  2. Widefield (CoolLED) dichroic mirror: 350-700 nm
Image Splitting Dichroic
  1. DM A561 nm long pass (FITC / TRITC-dual camera)
  2. Glass (transmits to camera 2)
  3. Mirror (reflects to camera 1)
Emission filter wheel camera 1 – transmitted (Photometrics BSI)
  1. single bandpass: 447/60 nm  → DAPI / BFP
  2. single bandpass: 525/30 nm  → GFP, Venus, mNeonGreen, Alexa488, FITC
  3. single bandpass: 600/52 nm → TRITC, mCherry
  4. single bandpass: 708/75 nm → Cy5
  5. quad-band 440/521/607/700 nm → DAPI / FITC / TRITC / Cy5
 Emission Filter wheel Camera 2 – relected (Photometrics BSI)
  1. single bandpass: 447/60 nm  → DAPI / BFP
  2. single bandpass: 525/30 nm  → GFP, Venus, mNeonGreen, Alexa488, FITC
  3. single bandpass: 600/52 nm → TRITC, mCherry
  4. single bandpass: 708/75 nm → Cy5
  5. quad-band 440/521/607/700 nm → DAPI / FITC / TRITC / Cy5
Cameras:
  • 2 x Teledyne Photometrics BSI (USB3) sCMOS camera, 95% quantum efficiency; 6.5 µm x 6.5 µm pixel area
Computer (HP Z8 Workstation): 

Dual Intel Xeon 4114 2.2 2400 MHz 10 CPU

  • RAM: 128 GB DDR4 2666 MHz
  • Nvidia Quadro RTX5000
  • 2 TB SSD drive for fast imaging
  • 6 TB HDD
  • 10 GB Ethernet card
System manual and additional material is available on SeaFile (login required).