Nikon X-Light-V3-03

Posted onAuthorGabriel Krens

Nikon X-Light-V3-03
Nikon X-Light-V3-03 - Image 2
Nikon X-Light-V3-03 - Image 3
Nikon X-Light-V3-03 - Image 4
Nikon X-Light-V3-03 - Image 5

The Nikon/Crest Optics X-light-V3-03 is a customized spinning disk imaging platform that offers a unique large Field Of View (∅ 25mm) Spinning Disk microscope, and a wide variety of objective dedicated for (extra-) large sample imaging. The triggered dual-camera configuration provides the optimal configuration for fast acquisition of large samples. The stage has an extra-large range and is equipped with an 8-slide holder, and additional custom  stage inserts to facilitate large sample mounting of cleared and expanded tissues samples. An in-house developed “serial block-face imaging” modality is currently under construction, and is to be implemented at a later time. Live imaging can be facilitated “on demand”, especially for aquatic model organism that benefit of water dipping objectives.

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Description

*coming soon*

Location: room i06.U1.406

Specifications
  • Nikon Ni-E upright microscope
  • Crest Optics X-light V3 spinning disk confocal
  • Dual-camera:  Photometrics Kinetix Back-Illuminated sCMOS (10.2 megapixel sCMOS – 6.5μm Pixels)
  • Motorized Focusing  nosepiece Up/Down movement (Up 13 mm/Down 2 mm)
    • Additional motorized Objective Piezo (Nano-F200N ) – 200 micrometer range (pending)
  • X/Y stage (Marzhauser) with 225 x 76 mm travel range:
    • i) 8 objective slides,
    • ii) 1-3 well-plates,
    • iii) X-large sample holder (under development)
  • in development: blockface imaging module (under development)
Light sources
  • CoolLED pE-300 for epi-fluorescence imaging: 395nm, 460nm, 575nm
  • Omicron BHU, with 4 independent laser lines: 405, 488, 555, 638 nm
Imaging modalities/features
  1. Widefield: LED-based
  2. Widefield: laser-based (trough X-light-V3 confocal head)
  3. CrestOptics X-light-V3-03 spinning disk offers a unique large Field Of View (∅ 25mm)
  4. Feedback microscopy (Nikon JOBS & Macro)

Nikon Ni2 – Widefield

  • Nikon Ni-E, upright automated microscope
  • CoolLED pE300 White MB
Filters cubes:
    1. DAPI: EX 392/25, DM 409, BA 447/60
    2. GFP/FITC: EX 474/27, DM 495, BA 525/50
    3. mCherry/TRITC:EX 554/23, DM 573, BA 609/54
    4. empty
Objectives: 2 position objective holder

  1. CFI Plan Apochromat Lambda D 4X / NA0.2 / WD 20mm (MDR70040)

  2. Nose-piece with 200 micron piezo, with option for following objectives:
Image Name Mag. Type Immersion NA Working distance  Cover glass Thickness Application Immersion value1
CFI Plan Apo
10XC Glyc		 10X Cleared / expanded Tissue Oil, Water Immersion, Glycerin, Silicone Oil 0.5 5.5 mm 0–0.17 Dipping LWD MRD71120 €€€€
CFI75
LWD 16X W		 16X Water 0.8 3.0 mm 0 Dipping LWD MRP07220 €€
CFI Plan Apochromat
Lambda D 20X		 20X Plan Apochromat Air 0.8 0.8 mm 0.17 MRD70270 €€
CFI Plan Apo LWD Lambda S 20XC WI 20X

Plan Apochromat

 Water 0.95 0.93 mm 0.11-0.23 Long Working Distance

MDR77205

€€€€€
CFI75 Apo 20XC W 20X Apochromat Water 1 2.8 mm 0–0.17 Dipping MRD77226 €€€€€€
CFI90 20XC Glyc 20X Cleared Tissue Glycerin 1 8.2 mm 0 Dipping MRD71220 €€€€€€
CFI75 Apo 25XC W 25x Water 1.1 2 mm 0–0.17 Dipping

MRD77220

€€€€€

11x “€” means €5000,- (ex VAT): €€ high-value objective; €€€ very high-value objective; €€€€ extremely high-value objective

Additional high-end bookable objectives (book in PPMS)
  1. CFI Plan Apo 10XC Oil, Water, Glycerin, Silicone / NA 0.5 / WD 5.5 mm (MRD71120) – €€€€ → suited for X-large sample / Extra-long working distance 
  2. CFI Plan Apo 25XC Sil / NA 1.05 / WD 0.55 mm  (MRD73250) – €€€ → suited for live Imaging

X-Light-V3 Confocal Scanner Unit

Excitation Light: Omicron V3 BHU – High Power w despeckler:
  • 4 lines: 405 / 488 / 555 / 640 (∼1200mW per line)
Spinning Disk:  25 μm pinholes; 135 μm spacing disk

Emission filter wheel – single camera mode
  1. ND 1.3 – 5%T for WF
  2. Emitter: FF01-442/42-25 (DAPI)
  3. Emitter: FF01-520/35-25 (FITC)
  4. Emitter: FF01-595/31-25(TRITC)
  5. Emitter: FF02-685/40-25(Cy5)
  6. empty (for dual-camera mode)
  7. empty
  8. empty
Emission Dichroic Mirror Block – dual camera mode
  1. Empty (reserved for single camera mode)
  2. V3 dual cam – Di 560LP → reflected (cam 2): FF01-520/35-32 (FITC)  / transmitted (cam 1): FF01-595/31-32 (TRITC)
  3. V3 dual cam – Di 560LP → reflected (cam 2): FF01-520/35-32 (FITC)  / transmitted (cam 1): FF01-680/42-32 (Cy5)

Emission Light detection: dual camera

  • Dual Photometrics Kinetix Back-Illuminated sCMOS (10.2 megapixel sCMOS – 6.5μm Pixels)
  • camera 1: Kinetix 0 → transmitted camera (horizontal)
  • camera 2: Kinetix 1 → reflected camera / vertical camera)
Computer (HP Z8 G5 Fury Workstation): 
  • Dual Intel Xeon W7-3446  4.6 GHz 20 CPU
  • RAM:256 GB DDR5 4800 MHz
  • Nvidia Quadro RTX A6000 Ada 48Gb
  • 6 TB SSD drive for fast imaging
  • 25 TB HDD
  • 10 GB Ethernet card
System manual and additional material is available on SeaFile (login required) – pending

Additional information

Building

i06
Camera / Detector

sCMOS
Fluorescent Lightsource

Laser: 405±5 nm, Laser: 488±5 nm, Laser: 561±5 nm, Laser: 640±5 nm, LED: 470-475nm, LED: 640-650nm
System_Specification

Illumination: Fluorescence, Illumination: Bightfield, Automated Stage, Dual Camera (see detectors)
Technology / Application

Large Sample Imaging, Time-lapse recording, High-troughput Screening

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