This setup is custom-built around a Yokogawa CSU-10 spinning disk confocal head mounted on a Nikon TE2000 inverted microscope with PFS attachment for continuous Z-drift compensation during live-cell imaging. AOTF-controlled laser line selection from a Coherent Innova 70C Multi-Line KrAr ion laser is combined with rapid filter-based emission selection for sequential, or emission beam-splitting for simultaneous, dual-color imaging of GFP/RFP or CFP/YFP combinations of fluorescent proteins. The laser light source is shared with a TIRF illuminator for critical angle illurmination. A choice of a high resolution Hamamatsu Orca ER or a high speed, high sensitivity Photometrics Cascade 512B electron-multiplying CCD camera is available. X,Y,Z axes are motorized to allow for position marking and revisiting during long-term timelapse recordings. A Piezo-driven Z axis top plate allows for fast Z stack acquisition with all available objectives. Objectives range from low magnification dry objectives for specimen positioning to high NA oil immersion objectives for TIRF (including a 100x1.49 PlanApo) and can be heated for live-cell imaging. The setup is integrated and controlled through MetaMorph.