The Zeiss Elyra PS.1 Combi system

NIC Super-Resolution & LSM 780 Confocal

Localization in the Arabidopsis root of the P-type proton ATPase. Image by Miyoshi Haruta, Sussman Lab (Department of Biochemistry).
Localization in the Arabidopsis root of the P-type proton ATPase. Image by Miyoshi Haruta, Sussman Lab (Department of Biochemistry).

Our Zeiss Elyra PS.1 Combi system has both confocal imaging capability (LSM 780) and super-resolution imaging capabilities: SIM (Structured Illumination Microscopy) and PALM (Photo-Activation Localization Microscopy). This instrument can also image samples using variable angle excitation and TIRF (Total Internal Reflection Fluorescence) microscopy.

Our Zeiss Elyra PS.1 Combi was installed in the NIC in 2014. It was made possible by a generous anonymous donation to the Department of Botany along with support from many units within UW-Madison including the College of Letters & Science, the College of Agriculture and Life Science, School of Medicine and Public Health, and the Graduate School along with contributions from a number of individual departments.

For super-resolution, the inverted microscope has four laser lines (405, 488, 561 and 640nm) and five color filtersets for both PALM and SIM super-resolution imaging. It is equipped with two EM-CCD cameras, an Andor iXon3 897 camera for PALM imaging in TIRF mode and an Andor iXon3 885 camera for 3D SIM.

For confocal, six laser lines are available (405, 458, 488, 514, 561, and 633nm). It is also equipped with a 34 element internal GAsP detector (LSM 780) for single-molecule sensitive full spectral imaging over the visible spectrum and a transmitted light detector for DIC imaging.

Microscope optics include the 100x/1.46 alpha Plan-APO for TIRF, the Plan-Apochromat 63x/1.40 for SIM, and 100x/1.57 alpha Plan-APO for PALM, 40x/1.10 C-Apochromat for confocal in addition to 10x, 20x, and 63x (water) objectives for confocal imaging. Filter sets include a 4 laser line primary beamsplitter for optimal unmixing capability.

Additional capabilities are a joystick-controlled motorized stage, stage insert for temperature and CO2 control (Tokai Hit), and a fiber optic-coupled computer running Zeiss’ ZEN software capable of imaging the same sample using three different technologies in succession (confocal, SIM and PALM).

Mouse brain slice imaged using a confocal in the NIC. Sample from the Riters Lab (Zoology)
Mouse brain slice (Riters Lab, Department of Zoology) imaged using the tiling capability of our Zeiss LSM 780 confocal microscope.
Micrographs of plant cells