Over the last 30 years, confocal microscopy has emerged as a primary tool for biological investigation across many disciplines. The simplicity of use and widespread accessibility of confocal microscopy ensure that it will have a prominent place in biological imaging for many years to come, even with the recent advances in light sheet and field synthesis microscopy. Since these more advanced technologies still require significant expertise to effectively implement and carry through to analysis, confocal microscopy-based approaches still remain the easiest way for biologists with minimal imaging experience to address fundamental questions about how their systems are arranged through space and time. In this review, we discuss a number of advanced applications of confocal microscopy for probing the spatiotemporal dynamics of biological systems.
Keywords: Colocalization; Confocal microscopy; Deconvolution; Fluorescence Lifetime Imaging Microscopy (FLIM); Fluorescence correlation spectroscopy (FCS); Fluorescence lifetime in photobleaching (FLIP); Fluorescence recovery after photobleaching (FRAP); Hyperspectral imaging; Image analysis; Linear unmixing; Machine learning; Neural network; Resolution; Superresolution microscopy.