Our Zeiss LSM 980 Airyscan2 system was installed in the NIC in 2023 and funded through support from the University of Wisconsin-Madison Vice Chancellor for Research and Graduate Education and the College of Letters and Science along with contributions from Botany’s Newcomb Endowment and other departments and units on campus whose researchers use the NIC microscopes.
For confocal, the rear port LSM 980 scanhead has tunable fluorescence emission over the visible spectrum entering a 32-channel GaAsP spectral detector, 2 cooled multialkalai PMT detectors, and a transmitted light detector for brightfield imaging. Six solid-state lasers are available: 405, 445, 488, 514, 561, and 639nm.
For super-resolution, the Airyscan2 detector enables 120nm resolution in xy and 350nm in z. This detector works with all available lasers and can also be utilized as an additional GaAsP confocal channel. Our Airyscan2 can be set up for multiplexing to enable increased speed of data capture of up to 47 FPS.
Microscope optics include a Plan-Apochromat 63x/1.4 oil, a C-Apo 40x/1.2 water immersion objective in addition to 10x, 20x, and 2.5x. Filter sets include multiple primary beamsplitters for optimal unmixing capability.
AI Sample Finder allows fast acquisition of a low magnification reference image and then a quick overview of fluorescence signal using the 2.5x objective captured with the side port Axiocam 705 camera (5MP CMOS). AI then automatically identifies the sample carrier on the low-magnification view and outlines possible regions of interest for imaging. This overview can then be used to quickly navigate to the region of interest for higher magnification data capture and to define tiled regions to image a larger sample area in high resolution.
Software is ZEN 3.8 and includes z-stack, time lapse, photobleaching, photoactivation, spectral unmixing, colocalization, 3D rendering, and experiment designer. A second offline processing PC runs ZEN 3.8 desk software including Image Analysis 2D and 3D, Intellesis AI Package, FRAP/FRET analysis and Airyscan Joint Deconvolution.
Additional capabilities are an enclosure and hardware for temperature and CO2 control, a joystick-controlled xy stage and a Piezo insert frame for precise z control of the sample holder during data capture. Sample holders include those for dishes, slides, large coverglass, and multiwell plates.
