Nikon CSU-W1-01 SoRa+NIR

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

The Nikon CSU W1-01 SoRa+NIR spinning disk microscope is an advanced enhanced resolution imaging modality, and the platform of the 355 nm UV laser ablation system. It has a piezo drive for fast and precise imaging of 3D stacks, and is equipped with an incubation chamber and a CO2/O2 gas mixer and an automated water immersion dispensing system.

Imaging modalities/features

  1. Confocal (spinning disk)
  2. Widefield (CoolLED)
  3. Super resolution via optical Re-assignment (SoRa)
  4. NIR imaging (not available in SoRa mode)
  5. Feedback microscopy
  6. UV laser cutter v2

Specifications

  • Nikon Ti2E inverted microscope stand (double deck) with Perfect Focus System (PSF)
  • Automated Water Immersion Dispensing System
  • Yokogawa CSU-W1 dual-disk spinning disk unit with 50 μm pinholes disk & 50 μm pinholes with micro-lenses SoRa disk
  • Yokogawa SoRa with magnification changer for optimum resolution across different objective magnifications (SoRa modality)
    • 4.0x; 60x objective; 55×55 μｍ FOV (40x objectives also possible with limited resolution gain)
    • 2.8x; 100x objective; 48×48 μｍ FOV
    • 1.0x; normal spinning disk imaging
    • Resolution
  SoRa mode (optical only) SoRa mode + deconvolution
XY resolution 150 nm 120 nm
Z resolution 320 nm 300 nm
  • 2 Teledyne Photometrics BSI cameras
  • Motorized stages:
    1. Nikon TI2-S-SE-E XY stage with linear encoders: 100 nm XY resolution, 500 nm XY accuracy
    2. Mad City Labs Z piezo & linear stage encoder: travel range: 200 μm, 0.4 nm min step size
  • Incubation chamber: ISTA custom-built dark chamber with Life Imaging Services heating unit (25-42 ºC)
  • Ibidi CO2/O2 gas mixer

Light sources

  • CoolLED – pE-300 White Light LED source
  • UV laser: 355 nm pulsed laser – not for imaging
  • Visible lasers: Omicron LightHUB UltraN
    • 405 nm laser:  (120 mW) – Continuous wave
    • 488 nm laser:  (200 mW) – Continuous wave
    • 561 nm laser:  (150 mW) – Continuous wave
    • 638 nm laser:  (200 mW) – Continuous wave
  • NIR laser: Omicron LightHUB NIR
    • 785 nm laser:  (200 mW) – Continuous wave

Filters for visual/widefield examination (lower deck)

  • DAPI-5060C Filter Cube: Ex 377/50, DM 409, Emission BP 447/60
  • GFP-4050B Filter Cube: Ex 466/40, DM 495, Emission BP 525/50
  • mCherry-C Filter Cube: Ex 562/40, DM 593, Emission BP 640/75
  • Cy5-4040C Filter Cube: Ex 628/40, DM 660, Emission BP 692/40

Objectives / position

  1. CFI Plan Apo λ 10x air / NA 0.45 / WD 4 mm (MRD00105)
  2. CFI Plan Apo λ 40x air / NA 0.95/ 0.17-0.25 mm (MRD00405)
  3. CFI Plan Apo VC 60x WI/ NA 1.2 / WD 0.28-0.31 mm / Water – (MRD07602) – €€
  4. CFI Plan Apo λ 60x oil / NA 1.4/  WD 0.13 mm / oil (MRD01605) – €€
  5. CFI SR HP Plan Apo λ 100x Sil / NA 1.35/ WD 0.30 mm / Silicone oil (MRD73950) – €€€
  6. Free slot for bookable objectives
 
Additional high-end bookable objectives (book in PPMS)
  1. CFI Apo LWD λ S 20XC WI / NA 0.95 / WD 0.9-0.99 mm / Water (MRD77200) – €€€

Facility objective (UV test IOF)

CFI Plan Fluor 40x / NA 0.75 / WD 0.66 mm (MRH00401)

Airy unit (AU) calculation for most objectives available for Nikon 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 Nikon CSU-W1-01 system (100x/1.35 NA Si oil is bookable). 
  • 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

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
  3. Near InfraRed (NIR laser) dichroic mirror: 785 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 (Photometrics BSI)

  1. quad-band 440/521/607/700 nm → DAPI / FITC / TRITC / Cy5
  2. single bandpass: 447/60 nm  → DAPI / BFP
  3. single bandpass: 525/30 nm  → GFP, Venus, mNeonGreen, Alexa488, FITC
 Emission Filter wheel Camera 2 (Photometrics BSI)
  1. quad-band 440/521/607/700 nm → DAPI / FITC / TRITC / Cy5
  2. single bandpass: 447/60 nm  → DAPI / BFP
  3. single bandpass: 525/30 nm  → GFP, Venus, mNeonGreen, Alexa488, FITC
  4. single bandpass: 600/52 nm → TRITC, mCherry
  5. single bandpass: 708/75 nm → Cy5
  6. single bandpass: 835/70 nm → NIR

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).

Contact: Imaging and Optics Facility