TI Workbench is a software package that serves as a control and analysis center for cellular imaging and electrophysiological experiments. It is unique among general-purpose software packages where it integrates the control of cellular imaging and electrophysiological devices, as well as sophisticated data analyses, which provides superior usability in imaging experiments combined with electrophysiology. During the development over the last 20 years, the range of supported image acquisition devices has expanded from cooled charge-coupled device (CCD) cameras to multi-photon microscope systems. In this review, I outline the concept of TI Workbench together with its unique functions and features derived from ideas emerging during daily experiments in my own lab and in those of my collaborators over the last 20 years. TI Workbench includes standard functions required for time-lapse multicolor fluorescence imaging and electrophysiological experiments, in addition to specialized functions such as random-scan or conventional raster-scan two-photon microscopy packages and fluorescence life time imaging (FLIM) utilities. Data analysis modules, e.g. digital data filters for temporal waveforms of time-lapse image data and electrophysiology and for 2-D image data, and fluorescence correlation spectroscopy (FCS) analysis functions, are well integrated with data acquisition functions. A notebook function holds formatted text, graphs, image and movie data altogether, which are linked to the actual data files. TI Workbench uses Igor Pro software as a back-end output for publishing. In addition, TI Workbench imports several different formats of image and electrophysiology data, serving as a general-purpose data analysis software package.