3i Spinning Disc

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General information

  Yokogawa Spinning disc confocal based on inverted IX81 microscope with automatic XYZ stage, and live experiment support.

Application
  • Multi-channel confocal/widefield fluorescence imaging of fixed/live samples
  • Multi-channel scanning TIRF imaging of live samples
  • DNA breakdown, photo-bleaching and photo-activation imaging
Imaging Mode
  • Wide-field Fluorescence
  • Spinning Disc Confocal
  • Scanning TIRF
Accessibility
Microscope details
  • Microscope frame: Olympus IX81 inverted microscope
  • Objectives: 20x/0.5, 40x/0.95, 60x/1.35 Oil, 100x/1.4 Oil, 100x/1.49 Oil TIRF
  • Light source for wide-field fluorescence illumination: X-Cite LED
  • Light source for confocal illumination: 3i 4-channel laser module (nm) - 405, 488, 561, 633
  • Bright-field imaging mode: BF, DIC
  • Spinning disc unit: Yokogawa CSU10
  • Auto-focusing: N/A
  • Live experiment support: heated live chamber (37 deg.) and 5% CO2 (humidified)
  • Camera
    • Confocal/TIRF imaging: Photomatric Evolve 512 EMCCD (512X512, 16um, 16 bits, 33.7 fr/s)
    • Widefield imaging: Hamamatsu ORCA ER (1344X1024, 6.45um, 12-bit, 8.3 fr/s)
  • Sample holder: universal slide/dish, 96-well plate
  • Optical bench: passive
  • Commercial dervice contract: 3i System Maintenance Agreement Class 200, - June 2021
Filter settings
Confocal/TIRF setup
Fluorophores DAPI GFP Cy3 Cy5 BF DIC
Laser 405 488 561 640 Halogen
Default filter cube 405M/488M/561M/640M (Pos2)
BS: 457/528/617/700, Em: quadband
BF (Pos1) DIC (Pos7, Analyzer)
Alternative emission filter 405a (Pos3, Em: 457nm) 488a (Pos4, Em: 528nm) 561a (Pos5, Em: 617nm) 640a (Pos6, Em: 700nm)

Widefield setup
Fluorophores DAPI GFP Cy3 Cy5 BF DIC
LED 377/50 485/20 560/25 650/13 Halogen
Default filter cube 405M/488M/561M/640M (Pos2)
BS:457/528/617/700, Em: quadband
BF (Pos1) DIC (Pos7, Analyzer)
Alternative emission filter 405a (Pos3, Em: 457nm) 488a (Pos4, Em: 528nm) 561a (Pos5, Em: 617nm) 640a (Pos6, Em: 700nm)


Workstation and software
  • Acquisition Software: Slidebook 6 (64-bit)
  • Workstation

    • CPU: Intel Xeon E5-2609
    • OS: Windows 10
    • RAM: 16GB
    • Local data drive: 1TB
    • Network name: cssd007397.ad.susx.ac.uk
Images taken from 3i
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