The spinning disk confocal with 3D-FRAP system is a multi-functional microscope built on a motorized Nikon Eclipse Ti inverted microscope equipped with a stage-top incubator and CO2 control system. It is capable of multi-position 2D and 3D live cell imaging in both spinning disk confocal and widefield modes.
Compared to laser scanning confocals, spinning disk systems acquire confocal images at higher speeds with less photodamage to the sample, which makes them ideal for live-cell imaging. The Yokogawa CSU22 confocal scanning unit on this microscope allows for spinning disk confocal imaging in two colours (green and red emission) and DIC, with images captured by a Photometrics Evolve EMCCD camera. The additional iLas 3D-FRAP module allows users to carry out high precision controlled photokinetic experiments (405/491/561 nm) during spinning disk imaging.
Widefield fluorescence, DIC and phase contrast imaging can be done on this microscope system using the second detector, a Hamamatsu ORCA Flash4.0 sCMOS camera. For both widefield and spinning disk confocal modes, a piezo z-drive enables fast acquisition of z-stacks for 3D imaging, and the Nikon Perfect Focus System (PFS) ensures long-term timelapse imaging without z drift. Sample holders for 35 mm diameter dishes and chamber slides (e.g. Ibidi, LabTek) are available. All imaging modes are controlled with MetaMorph software, which also allows for the review and analysis of acquired images.