Imaging facility of IEB CAS (http://www.ueb.cas.cz/if) has been officially classified as independent unit within the organization structure of IEB CAS at the end of 2015. However, its existence has started already in 2006 within the framework of research centre of MEYS REMOROST, project LC 06034 by the installation of confocal laser scanning microscope Zeiss LSM 5 Duo. Using this microscope, in 2006-2010 there have been published numerous original contributions. In 2009, the facility was equipped with automated station for in situ hybridization and immunohistochemistry on whole mounts and sections (OPPC project CZ.2.16/3.1.00/21159; 2009), which allowed improvement of immunofluorescence methods. The attention was paid as well to non-invasive in vivo fluorescence microscopy, which was extended by the installation of spinning disk confocal microscope Nikon Eclipse Ti-E with Yokogawa CSU-X1 (investment of CAS; 2012). This microscope opened new possibilities in the imaging and quantifying cellular events in higher spatio-temporal resolution. This trend has been extended in 2015, when new inverted Zeiss LSM 880 with sensitive spectral detection has been installed (OPPC project CZ.2.16/3.1.00/21519; 2015). The spatial resolution limit of this microscope was further improved by the installation of Airyscan detector (OPRDE project CZ.02.1.01/0.0/0.0/16_013/0001775; 2018). Recent highlight of the facility is a new LSM880 with Airyscan in horizontal setup, designed for plant biology needs (OPRDE CZ.02.1.01/0.0/0.0/16_013/0001775; 2018). Together with the upright microscope with structured illumination, Zeiss Apotome 2 (investment of IEB ASCR; 2015), the imaging facility is now optimized namely for demanding observations on living plant material, e.g. studies of dynamics of cytoskeleton, integral and peripheral plasma membrane proteins, plasma membrane associated-tethering complexes and lipids.