Facilities and Major Equipment

Department of Physiology and Cell Biology

Departmental Core Facilities

Confocal Microscopy

The confocal microscopy suite contains a Zeiss LSM 510 Meta laser confocal microscope, a Bio-Rad MRC 600 laser confocal microscope, and a Noran Odyssey laser confocal microscope. Also available within the Department is a high-speed Nipkow dual spinning disk confocal laser imaging system, consisting of a QLC100 high-speed confocal scan head, XR-Mega-10 ICCD camera and appropriate lasers for calcium imaging (See also: Calcium Imaging)

Transmission Electron Microscopy

This facility houses a Phillips CM10 transmission electron microscope that is equipped with an analog negative (5x7) photosystem as well as a Gatan digital imaging system (BioScan 792). The adjacent rooms contain a fully equipped darkroom and two Leica (Ultracut S) microtomes with Reichert FCS cryo attachment for cutting frozen sections for electron microscopy. Other microtomes are available for tissue preparation and light microscopy.

Molecular Core

This facility is fully staffed and equipped with shared instrumentation that includes real-time quantitative PCR (Applied Biosystems ABI 7300 sequence detector), gel imaging apparatus (Bio-Rad Molecular Dynamics PhosphorImager), and four 96-well PCR thermal cyclers, Additional instrumentation includes gel electrophoresis, rocking incubator, hybridization chamber, electroporation units, centrifuges, refrigerators and freezers, and software for sequence analysis.

Tissue Culture

The fully staffed Tissue Culture Facility contains primary and cell-line-dedicated incubators, high- and low-speed centrifuges, dissecting microscopes, biosafety level II hoods and safety cabinets, fluorescent inverted microscope with Q-imaging, and cryogenic (liquid nitrogen) storage.

Histology and Immunocytochemistry

A workstation for paraffin wax-embedding, sectioning (rotary microtome), collection of sections, and staining, as well as a Nikon microscope equipped with a z-axis encoder and drawing tube is available for histology needs as well as design-based stereological quantification of particles in tissue sections. In addition, a motorized cryostat (Leica CM3050) and vibratome is available along with upright and inverted fluorescent microscopes with digital imaging systems. Wet laboratory with dissecting microscopes for tissue processing is also available.

Calcium Imaging

A workstation to visualize organelle traffic and rapid calcium waves in cells, using a high-speed Nipkow dual spinning disk confocal laser imaging system, consisting of a QLC100 high-speed confocal scan head, XR-Mega-10 ICCD camera, filter sets, fiber optics, 4-channel acousto-optical tunable filter (AOTF), with the scan head mounted on a Nikon E600 FN upright Physioscope with fluorescent water immersion lenses.

HPLC

An HPLC system with an electrochemical detector (Agilent Technologies) is available, as well as an HPLC system with a fluorescence detector (Agilent Technologies).

Cell Sorting

This Facility houses a 4-color Beckman Coulter XL/MCL analytical flow cytometer with a single 488 Ar laser, configured to measure emissions at 525 (fluorescein, AF488, CFSE, etc.), 575 (R-phycoerythrin), 670 (7-AAD, PerCP and PE tandems such as PE-cyanine 5) and >740nm (PE-cyanine 7 tandems). Propidium iodide (PI) can be effectively measured at 575, 670 or at longer wavelengths. Excellent analytical software (FlowJo, Tree Star, Ashland, OR) is available for use.

Computer Support

A full-time computer expert is available for all computer and software needs in the Department.

School of Medicine Core Facilities

Confocal Microscopy

The facility provides training and scoping time on the FV1000 Olympus laser scanning confocal microscope. This facility also houses two Olympus FV300s workstations for two-photon microscopy on upright and inverted microscopes.

Electronics/Technical Support

The School of Medicine provides a full-time staffed electronics and instrument repair facility, located within the Department of Physiology.

Transgenic Facility

The Core contains a gene targeting center that provides a complete ES cell culture and targeting service, a microinjection center, for both fertilized eggs and blastocyst injections, and an animal service that provides animals and embryos for the center, performs embryo transfers, and has responsibility for managing transgenic founders and colonies. Additional services include genotyping, karyotyping, breeding mice and cryopreservation services, assay ES cell clones, target construction. For details, see www.unr.edu/med/dept/pharmacology/transgenic/

University of Nevada, Reno (UNR) Core Facilities