This workstation is only available to users already trained on the Zeiss LSM880 microscopes. The PC is a Hewlett-Packard Z6 workstation with an 8 core Intel Xeon CPU, 64GB of RAM, 12TB of data storage (RAID 10), running Windows 10 (64 bit) with a licensed version of the Zeiss ZEN software for data processing (stitching, spectral unmixing, etc.) and FIJI (ImageJ).

Programmable microwave tissue processor, with continuous power settings from 100-750 Watts, load cooling to maintain consistent temperatures, with built-in fume extraction and vacuum.

The microwave is useful for:

• improved tissue fixation
• improved retention of fluorescence after fixation
• for more rapid tissue processing for immunocytochemistry
• electron microscopy
• decalcification.

The Zeiss LSM 880 NLO upright multiphoton/confocal microscope includes a 34-channel detector for spectral information and additional non-descanned GAsP detectors. The system includes 7 visible spectrum laser lines (405, 458, 488, 514, 561, 633 nm), plus an additional software-tunable Spectra Physics Mai Tai HP DeepSee laser (690-1040 nm). The system includes software modules for controlled region-of-interest scanning, deconvolution, physiology experiments, performing FRAP & FCS and control of the motorized stage to allow stitching together images from multiple fields of view into one larger image (montage). The software also enables visiting multiple sites during a time lapse acquisition, and the instrument includes an environmental chamber suitable for live cell imaging at a variety of temperatures. Wet lab space is available to users of the facility in an adjacent room, including cell culture incubators and a biosafety hood.

NOTE: BSL2 samples must be discussed with the on-site core manager BEFORE they arrive in the facility. Live animals - invertebrates only!

The Fast Airyscan allows for resolution improvement up to 1.9x of typical confocal resolution and the fast mode sacrifices a bit of resolution for speed (useful for live cell imaging). The scanhead includes a 34-channel detector for spectral information.

The Zeiss LSM 880 inverted confocal microscope includes a 34-channel detector for spectral information, and 7 Laser lines (405, 458, 488, 514, 561, 594, 633nm). The system includes additional software modules for performing FRAP and control for the motorized stage to allow stitching together images from multiple fields of view into one larger image (montage), as well as visiting multiple sites during a time lapse acquisition. The instrument includes an environmental chamber suitable for live cell imaging at a variety of temperatures, humidity and CO2 capabilities. The facility has wet lab space, cell culture incubators, and a biosafety hood in an adjacent room.

NOTE: BSL2 samples must be discussed with the on-site core manager BEFORE they arrive in the facility.

Objective lenses (with matching DIC prisms) include:

• Plan-Apochromat 10x/0.45 (WD=2.0mm)
• Plan-Apochromat 20x/0.8 (WD=0.55mm)
• Plan-Apochromat 40x/1.3 Oil
• Plan-Apochromat 63x/1.4 Oil
• Plan-Apochromat 40x/0.95 dry CORR (not typically on the microscope, available upon request)

This workstation is only available to users already trained on the Zeiss Elyra S.1. First priority is given to users scheduled on the Zeiss Elyra S.1 for processing their data. The PC is a Hewlett-Packard Z6 workstation with an 8 core Intel Xeon CPU, 64GB of RAM, 12TB of data storage (RAID 10), running Windows 10 (64 bit) with a licensed version of the Zeiss ZEN software for data processing and FIJI (ImageJ).

The Zeiss Elyra S.1 uses the technique of structured illumination (SR-SIM) to achieve resolutions up to two times better than that of a confocal, deconvolution or standard fluorescence microscope. Resolutions in XY of 110nm and in Z of 300nm are possible (resolution is wavelength and sample-preparation dependent), allowing users to image a volume in the sample that is 1/8 that of previous techniques.

The good news is that this super-resolution technique is a not-too-difficult jump from the dyes and sample prep techniques used for a confocal or deconvolution microscope. The Elyra S.1 can image four fluorescent colors, roughly corresponding to DAPI, FITC/GFP, Rhodamine/RFP, and CY5 (see below). SR-SIM can image to depths of 10-15 microns into a sample. Note: SR-SIM does not work well (or at all) with dimly fluorescent samples or samples that photo-bleach.

The Elyra S.1 has the following microscope objectives:

• Plan-Neofluar 10x/0.30
• Plan-APO 40x/1.4 Oil
• Plan-Apochromat 63x/1.40 Oil
• Plan-Apochromat 63x/1.30 Oil/Water/Glycerine KORR
• Alpha Plan-APO 100x/1.46 Oil

The laser lines are 405, 488, 561, and 642nm. Imaging can be performed somewhat faster using a single multi-band cube, or with 4 sequential cubes to reduce spectral overlap:

• EF LBF 405/488/561/642
• EF BP 420-480 / BP 495-550 / LP 655
• EF BP 495-550/BP 570-620
• BP 420-480/LP 655