Nikon X-Light-V3-01 + OT

Location: room i04.U1.019 – book in PPMS

System description and specification

The Nikon/Crest Optics X-light-V3-01+’Optical Trap’ offers a tissue rheology probing platform in combination with Spinning Disk microscope, with a broad range of excitation lines throughout the visible spectrum. The dual-camera system configuration provides is optimized for fast acquisition for live processes and/or large samples size. The z-piezo stage provides fast and precise imaging of 3D stacks. The automated water immersion system allows prolonged and combined live imaging experiments.

 The automated water immersion system allows prolonged and combined live imaging experiments. This incubation chamber and CO2 mixer with humidifier makes this system best suited for live imaging.

The Optical Trap enables measuring rheoglogy, forces and interactions in a direct, absolute and non-invasive way in the field of Cell Mechanobiology, Molecular Biophysics, Phase Separation and Soft Matter Physics applications.

Specifications
  • Nikon Ti2E inverted microscope
  • Crest Optics X-light V3 spinning disk confocal
  • dual-camera: Hamamatsu Orca FusionBT  (5.3 megapixel sCMOS – 6.5μm Pixels)
  • Tube-port camera: Hamamatsu Orca Flash LT3 (Optical Trap & Widefield)
  • 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, 0.4 nm min step size
  • Perfect Focus System (PSF4) – PM compatible (Optical Trap)
  • Automated Water Immersion Dispensing System.
  • Climate control: heated incubator + CO2 mixer + humidifier
Specification Optical Trap / Tweezers sources

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Light sources
  • CoolLED pE-300 for epi-fluorescence imaging: 400nm, 460nm, 550nm
  • Lumcore CELESTA + Despeckler, with 7 independent laser lines for confocal imaging: 405, 446, 476, 518, 545, 637, 748 nm (~800 mW per channel)
  • Toptica Photonics IR trapping laser: 1064 nm, 5W power (Laser Safety Class 4)
Imaging modalities/features
  1. Widefield (LED-based): large FOV, using the Nikon camera
  2. Crest Optics X-light-V3-02 spinning disk offers a unique large Field Of View (∅ 25mm)
  3. Widefield (laser-based)
  4. Feedback microscopy (Nikon JOBS & Marco)

Ti2 – Widefield

  • Nikon Ti2E, with automated water immersion
  • CoolLED pE300 White MB

Filters 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-D 10x air / NA 0.45 / WD 4 mm (MRD701170)
  2. CFI Plan Apo λ-D 20x air / 0.8/ 0.8 mm (MRD701170)
  3. CFI Plan Apo λ 40x WI / 1.15/  0.60 mm (MRD77410) – €€€
  4. CFI Plan Apo VC 60x WI / NA 1.2/  WD 0.31 mm (MRD07602) – €€ – Optical Trap
  5. CFI Plan Apo λ 60x oil / NA 1.42/ WD 0.15 mm / oil (MRD71670) – €€
  6. empty
 
Additional high-end bookable objectives (book in PPMS)
  1. CFI P-Apo 10X MuliCorr / 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

X-Light-V3 Confocal Scanner Unit

Excitation Light:
  • Lumcore CELESTA: 405, 446, 476, 518, 545, 637, 748 nm (~800 mW per channel)
  • 18mmHT micro-lenses
Spinning Disk 
  • Standard pattern → 50 μm pinholes; 250 μm spacing disk

Emission filter wheel – single camera mode
  1. Emitter: FF02-438/24 (DAPI)
  2. Emitter: FF01-511/20 (FITC)
  3. Emitter: FF01-595/31(TRITC)
  4. Emitter: FF02-685/40(Cy5)
  5. Emitter: FF01-819/44 (Cy7)
  6. Emitter: FF02-485/20-25 (CFP)
  7. Emitter: FF01-560/25-25 (YFP)
  8. Empty

Emission Dichroic Mirror Block – dual camera mode

    1. Celesta DAPI/Cy3-GFP/Cy5 DualCam Set (32mm)

    2. Celesta CFP/YFP Dual Cam Set (32mm)

    3. empty (transmitted light)

Optional: Celesta GFP/mCherry Dual Cam Set (32m)

Emission Light detection: dual camera

      • 2 x Dual Hamamatsu Orca FusionBT  (5.3 megapixel sCMOS – 6.5μm Pixels)
      • camera 1: transmitted camera (horizontal);
      • camera 2: reflected camera / vertical camera)
 
Computer (HP Z4 High-Data Workstation): 
  • Dual Intel Xeon W-2245  3.9 GHz 8 CPU
  • RAM: 128 GB DDR4 2666 MHz
  • Nvidia Quadro RTX A5000
  • 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).