Confocal Spinning Disc 3 Nikon Ti2 – Yokogawa X1

Spinning-Disc-3
Suzan Ruijtenberg and Mike Boxem are imaging the worm (Ceanorhabditis elegans), in which their newly developed CRE-loxP based recombination system is illustrated in the intestine. Tissue specific expression of CRE leads to an intestine specific knock-out of their gene of interest in blue and, at the same time, causes a switch from red to green in the intestine while other tissues are unaffected.

Primary tasks:
Confocal Spinning Disc microscopy, 3D imaging

Microscope: Nikon Eclipse Ti2-E (motorized) with perfect focus

Stage: Ti2-S-SE-E (motorized)

Confocal Head: Yokogawa CSU-X1-M1

Piezo: MCL Nano-Z100-N2

Objectives:

  • Nikon CFI Plan Apo λ 10x/0.45 4.0mm WD 556 (Prime BSI) / 703 nm/px
  • Nikon CFI Plan Apo λ 60x/1.40 0.13mm WD OIL 94 (Prime BSI) / 118 nm/px
  • Nikon CFI Plan Apo λ 100x/1.45 0.13mm WD OIL 71 (Prime BSI) / 57 nm/px

Omicron LightHUB 6 with lasers:

  • 405 0.91 mW
  • 488 4.48 mW
  • 561 3.18 mW
  • 642 9.59 mW

Epifluorescence light source: CoolLED pE-300 White MB (LLG)

Filters:

  • BFP [EX405/10 / FF416]
  • GFP-A-Basic [EX469/35 / DM497 / BA525/39]
  • mCherry-C [EX562/40 / DM 593 / BA640/75]

Emission filter wheel: Sutter LB10-B wheel with:

  • 496LP; 525/30
  • 617/73
  • 512/630 dual
  • ET460/50
  • ET705/72

Detectors: Teledyne Photometrics Evolve EMCCD (512x512px) & Prime BSI sCMOS (2048x2048px)

Temperature control and CO2: Tokai Hit STX stage top incubator

Software: MetaMorph 7.10

Installation: 2009 (original), 2022 (extensive upgrade)

Location: O-519
Contact Person: Ruben Schmidt