Cellular and Molecular Imaging Core

The mission of the Cellular and Molecular Imaging (CMI) Core facility of the Integrative Neuroscience COBRE center is to provide infrastructure and resources necessary to perform cellular and molecular imaging of biomolecules as well as media preparation to further research at the University of Nevada, Reno (UNR). The Core facility is located on the third floor of the Fleischmann Lifesciences building in the Biology Department of the College of Science.

The CMI Core facility provides access to state-of-the-art instruments including a new Scientifica HyperScope two-photon imaging system for deep tissue scanning in live brains and whole animals, a Leica SP8 confocal microscope system equipped with a LIGHTNING allowing the capture of cellular details and observe dynamics with a resolution down to 120 nm, a Leica Thunder 3D Tissue upright microscope, a Leica Thunder Model Organism stereomicroscope intended to image embryos, organs and small organisms such as Drosophila and C. elegans, a GE Phosphoimager for Northern blotting, a BioRad ChemiDoc for Western blotting, two BioRad Real-Time PCR instruments for precise and quantitative PCRs, a QIAgility for automated PCR set-up, a GeneSys 180 UV/Vis spectrophotometer, and a Quibit 4 fluorometer.

Further capabilities of the core includes media preparation instruments and resources for husbandry of Drosophila and C. elegans used as model organisms.

Location: FA 318 (door key access required)

Two-photon microscopy has become an indispensable tool for cellular neuroscience, allowing high-resolution fluorescent imaging of cells in vitro and in vivo living brain and whole animals. The HyperScope system is a custom-build, two-photon microscope equipped with a Mai-Tai eHPDS broadly tunable Ti-sapphire laser (690-1040 nm), multiple objectives (16x, 20x and 40x), and stands on a Thorlabs breadboard and anti-vibration table. With the Scientifica SciCam Pro CCD camera, it is possible to perform functional imaging of video frame rates of 24 fps (full resolution) and 40 fps (binned 2 x 2). The detector assembly includes one red-shifted PMT and one GaAsP PMT modules, and epifluorescence imaging with filter sets for GFP (470/40x extinction, 525/50m emission) and mCherry (560/40x extinction, 630/75m emission). The scan mirrors in the imaging path can be arranged in multiple configurations depending on your needs. With up to 3 mirrors in series, you can choose between galvo/galvo (GG), resonant/galvo (RG), or resonant/galvo/galvo (RGG) arrangements. The microscope is fitted with an ultra-smooth Z-axis drive to ensure vibration free movement, with the Z-axis being controlled with software for acquiring z-stacks with a minimum step size of 0.1 mm. The addition of a Piezo z-axis objective positioner allows volume imaging up to 400 mm. The microscope can be easily configured and switched between in vitro and in vivo applications by removing the substage optics and riser block. The system is enclosed in a black-out cage to prevent light and noise pollution of biological samples.

This platform includes an award winning Scientifica SciScan acquisition software, allowing users to integrate SciScan easily with custom-written applications, such as LabView and ScanImage. The ScanImage software interface (Vidrio Technologies) used by Scientifica was developed by HHMI Janelia Farm specifically for neuroscience applications, and is considered to be the most advanced software for two-photon laser scanning on the market.

Key unique features of the Scientifica HyperScope include:

Location: FA 324

The Leica TCS SP8 LIGHTNING confocal allows image information extraction with super resolution acquisition that exploits the sub-diffraction lateral resolution of confocal microscopy. It allows to capture cellular details and observe dynamics with a resolution down to 120 nm, which is close to STED super resolution microscope capabilities. The upright confocal is capable of simultaneous collection from up to 3 detection channels (two PMTs, one HyD) using multiple laser lines (diode 405, blue 488, green 552, red 638 nm). The confocal is configured with a conventional tandem scanner (8 kHz), a super Z-galvo stage type S, multiple objectives (10x, 20x, 25x, 40x, 63x), and stands on an anti-vibration table. The confocal is connected to a computer workstation equipped with Leica LAS-X software and a 30 inch monitor.

Key features of the confocal include:

Location: FA 324

THUNDER imagers are a brand-new class of computational clearing high-resolution wide-field microscopes developed exclusively by Leica Microsystems. These imagers can acquire sharp data and high-quality images by instant removal out-of-focus blur or haze inherent to thick 3D samples that are generally acquired using widefield microscopy. These imagers also significantly shorten the imaging workflow from image acquisition to data analysis with the new LAS-X navigator software. The THUNDER Imager 3D Tissue is a motorized upright microscope and is outfitted with a ultra-fast DFC 9000 GT/C camera, and is ideal for cell culture, tissues, fixed samples, and cleared tissues. The THUNDER Imager Model Organism is a M205 FCA stereomicroscope intended to image embryos, whole organs and small organisms.

Key features of the THUNDER 3D Tissue imager include:

Key features of the THUNDER Model Organism imager include:

Location: FA 312 (Shared Core/Equipment room, door code access required)

The Typhoon Phosphoimager is a fast and versatile laser scanner for sensitive and quantitative measurements of radio-isotropic-labeled samples. The Typhoon is a non-confocal variable mode laser scanner offering high-speed and resolution for precise quantitation of proteins, nucleic acids and other biomolecules. The scanner comprises "filmless" autoradiography with storage phosphor screens. The laser can generate 16-bit images at up to 25 μm pixel resolution to give precise quantitation in gels and blots, with a high speed to scan gels of up to 240 x 250 mm in less than 2 min. Two phosphor screens with a size of 200 X 250 mm are available for autoradiography. The scanner can detect a broad range of isotopes in radiolabeled samples. Directly adjacent to the phosphoimager system, there is a PC computer with a software package that allows on-site quantitative and qualitative analysis of images and data.

Key features of the Typhoon imager include:

Location: FA 312 (Shared Core/Equipment room, door code access required)

The Biorad Chemidoc is an easy, high-performance gel and blot imaging for accurate quantification of protein samples in gels and membranes. The Chemidoc streamlines Western blotting in a five-step approach by combining traditional blotting techniques with innovative tools to allow improved band quantitation and quick assessment of electrophoresis results and blot transfer quality prior to western blotting. The Chemidoc enables rapid western blot results in as little as 15 min with efficient protein transfer in 3 min, and 5 min stain-free gel imaging. The imbeded software includes all the tools for quantitative and qualitative image and data analysis. Images taken on the ChemiDoc can also directly be print.

Key features of the ChemiDoc system include:

Location: FA 314 (Tissue Culture facility, door code access required)

The Trans-Blot Turbo transfer system combines traditional blotting techniques with modern filter paper and buffers allowing rapid transfer of proteins from gels to membranes with minimal preparation time. The system integrates a high amperage power supply that directs current between a built-in platinized titanium anode and a stainless steel cathode.

Key features of the Turbo-Blotter include:

Location: FA 313 (Autoclave room)

The Integra media autoclave allows the rapid and gentle preparation of 1-10 liter culture medium. Precise controlling of temperature, time and pressure during the sterilization process guarantee constant high quality. The mediaclave can be used for the preparation and sterilization of numerous types of culture media, including media for C. elegans research. The wide adding port facilitates safe addition of supplements prior to dispensing. The mediaclave is connected to a peristaltic pump housed in a vertical flow workstation to fill petri dishes and vials.

Key features of the Mediaclave include:

Location: FA 312 (Shared Core/Equipment room, door code access required)

Two Biorad real-time PCR machines combine advanced optical technology with precise thermal control to deliver sensitive, reliable detection, quick set-up runs and monitoring of amplification traces in real-time on the integrated LCD touch screen. Directly adjacent to the two qPCR detection systems, there is a PC computer containing easy to use software for on-site analysis of qPCR data.

Key features of the CFX96 include:

Location: FA 312 (Shared Core/Equipment room, door code access required)

The QIAgility is a compact benchtop instrument that enables rapid, high-precision setup of PCR experiments with ready-to-use software for ease and convenience. The software provides step-by-step guidance for worktable setup and automatic calculation of all mixes, eliminating the need to program pipetting steps. Just select the kit and cycler, define your targets and start the run. For added flexibility the QIAgility is operated via a laptop computer.

Key features of the QIAgility include:

Location: FA 312 (Shared Core/Equipment room, door code access required)

The GeneSys spectrophotometer performs high-throughput quantitative UV-Vis measurements where a double beam is required as a reference cell position. Optimized for usability and performance, this system features a high-resolution color touchscreen, and optional Wi-Fi networking. The compartment is a 8-position cell changer (standard).

Key features of the GeneSys 180 UV/VIS include:

Location: FA 314 (Tissue Culture facility, door code access required)

The Qubit 4 Fluorometer accurately measures DNA, RNA, and protein quantity. The Qubit together with a RNA IQ Assay Kit can accurately distinguish intact from degraded RNA in just two easy steps. For all Qubit assays, the concentration or quality of the target molecule in the sample is reported by a fluorescent dye that emits a signal only when bound to the target, which minimizes the effects of contaminants on the result. The easy-to-use touch-screen menus make it easy to select and run the assays you need, with results displayed in just a few seconds.

Key features of the Qubit 4 Fluorometer include:

A Lab/Core Technician (tbd) on staff provides basic training and technical support in order to assist researchers with all of our equipment and software. For information about the core, please contact the core director Alexander van der Linden, Ph.D. at [email protected].

For confocal and two-photon imaging:

For molecular imaging and accessories: