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Spinning Disk Confocal Microscopes

The SCF houses several high-end, fully motorized spinning disk confocal microscopes. Each features at least 4 excitation laser lines, one or two spinning disk types, at least one highly sensitive sCMOS camera, and full environmental control.

Optimized for live-cell observation in 3D and/or at high speed/throughput, these microscopes offer an excellent compromise between imaging speed, sectioning capability, efficient light exposure, and sensitivity. Each system offers a fully equipped wide field transmission and epi-fluorescence capability in parallel, freely combinable with confocal observation channels.

The set-ups are particularly specialized for certain experiment types, often through the choice of microscope base, disk type, and (extra) peripherals. A brief overview follows below.

For detailed descriptions of the individual workstations (all specifications, filter settings, etc.) and interactive spectra viewers, see our internal Wiki (D-BSSE login required).

X1 Upright Spinning Disk Confocal

The X1 system is an upright microscope, specialized in high-speed live-cell observation for samples requiring in situ imaging on special sample holders or devices with a non-transparent bottom. The system is outfitted with full environmental control for live cell support (temperature, CO2, humidity) and a series of dry or water dipping lenses for direct, from-above immersion into live-cell environments, such as neurons growing on a MEA-chip or organotypic cultures on opaque surfaces.

For 3D observation, the microscope is equipped with the fastest spinning disk unit at the SCF (max. 10000 rpm = max. 2000fps). In most practical cases however, X1 fluorescence imaging is limited to a full frame rate of about 50fps, due to signal intensities and/or data-transfer rates from the camera. For improved illumination uniformity, the scanner is equipped with a Homogenizer unit (Visitron Systems).

In addition, for wide field imaging one can make use of an intensified high-speed CMOS camera (HiCAM Fluo) using a dedicated acquisition station. Geared toward capturing the fastest live-cell dynamics (e.g. neuronal signaling), this combined intensifier+detector can record full frame time-lapse sequences with a max. frame rate of 1000fps in any of the wide field fluorescence channels.

  • Microscope base: Nikon Ni-E (upright)
  • Confocal disk unit: Yokogawa CSU-X1
  • Uniformity unit: Homogenizer (Visitron Systems)
  • Disk format: 50 µm pinholes @ 250 µm spacing, dual disk technology
  • Available lasers: 405, 488, 561, and 640 nm
  • Camera at SD module: Photometrics Prime 95B (w. 2.5x relay lens)
  • Piezo z-stage: Mad City Labs Nano-Drive (200 µm)
  • Wide field light path: through disk (low throughput) or side-port (high speed)
  • Wide field excitation: Lumencor Spectra X (390/438/470/510/542/632 nm)
  • Wide field camera: Lambert Instruments HiCAM Fluo (high-speed camera)
  • Specialty: upright configuration, BSL2 experiments, ultra-high speed wide field imaging