Nikon Ti2-W1 spinning disk confocal microscope
The Ti2-W1 is a Nikon based spinning disk system using a Yokogawa W1 scanhead attached to a Ti2 inverted research microscope.
Nikon Ti2-Wi confocal microscope
image: Christian Ackermann
Spinning Disc Units from Yokogawa are very efficient because a second disk with microlenses is mounted opposite the disk with thousands of pinholes. These microlenses ensure that a large portion of the laser radiation passes through each pinhole and can be utilized for excitation. The images are then captured with CMOS or CCD cameras, which have a much higher quantum efficiency than the detectors of point scanning confocals. All these factors together make spinning disk systems particularly suitable for live imaging, as phototoxicity and photobleaching are greatly reduced. The W1 has two disks, one with 50 and one with 25µm pinholes, so that objectives with lower magnification can be used closer to strict confocality (e.g. 20x NA 0.95 water immersion objective: 4 A.U. with 50µm pinhole, 2 A.U. with 25µm pinhole).
The W1 system offers 6 laser lines: 405nm, 445nm (not functional at the moment), 488nm, 514nm, 561nm and 647nm. Image acquisition can be performed with single emission filters (high specificity) or in a triggered mode with fast laser switching through a 4-band filter (DAPI, FITC, TRITC, Cy5) or a 3-band filter (CFP, YFP, mCherry) - this mode is faster but less specific.
The camera installed is a Zyla 4.2P (CMOS camera, 2048 x 2048 pixels with a size of 6.5μm, QE up to 82%).
Additional features: On-stage environmental chamber from TokaiHit (temperature/CO2 control). Fast z-stacks can be obtained with piezo driven stage-insert (range 100μm), additional to the normal z-motor of the Ti2 microscope. Automatic water dispenser for experiments exceeding approximately 2 hours.
Objectives
- Nikon Plan Apo λ 10x NA 0.45 (WD 4mm)
- Nikon Plan Apo λD 20x NA 0.8 (WD 0.8mm)
- Nikon Apo LWD 20x WI λS NA 0.95 (WD 0.95mm)
- Nikon Apo LWD 40x λS NA 1.15 (WD 0.6mm)
- Nikon Plan Apo λS 40x SIL NA 1.25 (WD 0.3mm)
- Nikon Plan Apo IR 60x NA 1.27 (WD 0.16-0.18mm)
- Nikon Plan Apo λ 60x Oil NA 1.4 (WD 0.13mm)
- Nikon Plan Apo λD 100x Oil NA 1.45 (WD 0.13mm)
