The Harvard Center for Biological Imaging was opened May 17, 2010 and is one of the most innovative biological imaging centers in the world. Under the direction of Harvard faculty member Jeff Lichtman, HCBI Director of Imaging Doug Richardson, and in partnership with Carl Zeiss Microscopy; the HCBI is now recognized as a true “Evergreen” microscopy facility. In order to ensure investigators always have access to the latest and best technologies, the six most advanced imaging systems at the HCBI are replaced every 2-3 years, ensuring that the facility remains relevant and is never in possession of outdated equipment. Currently, the HCBI contains 16 microscopes and high-end processing stations including an ELYRA multi-function super-resolution system (LSM 710 confocal/TIRF/SIM/PALM/STORM), a LSM 780 upright confocal configured for 2-Photon imaging, a LSM 780 inverted confocal with environmental control, a LSM 700 inverted confocal, a fully environmentally controlled Cell Observer widefield microscope for long-term imaging, the AxioZoomV16 fluorescence zoom microscope, and an Axio Scan high throughput slide scanner. In addition, we recently installed the first commercial Lightsheet microscope for live cell imaging in North America. The facility also features wet lab space, eukaryotic and prokaryotic incubation, and access to nearby animal care facilities for a wide range of species. Further, Sven Terclavers, an embedded Zeiss specialist, is also present in the facility to assist with experimental design, advanced imaging configurations, training, and troubleshooting. All microscopes are covered under platinum service agreements to ensure rapid repair and minimum down times.