CNRM Scientific Cores

Microscopy Core

Facilities

Microscopy core has 2-Photon microscope/electrophysiology, Sterology, and Biolistic Transfection facilities at USUHS.

2-Photon Microscope/Electrophysiology

Location: USUHS B3131

Equipment:

• Microscope - Zeiss LSM 7 MP system with an upright Axio Examiner microscope and Coherent Cameleon pulsed Ti:Sapphire Laser. Operating system for the microscope is "Zen 2009" software. Filter cube combinations currently support detection of green fluorescence (GFP, Calcium Green, Alexa 488, etc.) and red fluorescence (e.g. Rhodamine dyes).
• Electrophysiology - Molecular Probes Axon CNS MultiClamp 700B two-channel amplifier and "pClamp 10" software for pulse generation and data acquisition. Additional equipment includes a GRASS S88K Dual output stimulator, a pico-injector for bolus loading of dyes or focal stimulation and an 8-channel peristaltic pump for bath perfusion. Best suited for imaging slices or in vivo preparations with concurrent electrophysiological measurements.

Scheduling

Contact Fritz Lischka for initial training and scheduling. An on-line scheduling system may be implemented in the future.

Points of Contact

• Dr. Fritz Lischka, Email
• Dr. Sharon L. Juliano, Core PI, Email

Sterology

Location: USUHS BIC G233

Equipment:

• Zeiss AxioImager M2 upright microscope and "Stereo Investigator" software package from MicroBrightField. The microscope is equipped with a wide range of objective lenses (2.5x - 63x), standard and oil immersion condensers, an X-Cite fluorescent light source, and filter cubes appropriate for imaging red, green, and blue fluorophores.
• The system also has a motorized stage and two digital cameras.
• The Zeiss Axiocam MRc is a color camera suitable for brightfield imaging, while the Hamamatsu Orca-R2 captures monochrome fluorescence images.
• The Stereo Investigator software allows for unbiased quantification of parameters such as cell numbers, lengths, areas, and volumes. The software also implements several statistical analysis methods.

Scheduling

Use the BIC on-line scheduling system, see user guide for instructions. Contact Dennis McDaniel for initial training.

Points of Contact

• Dr. Dennis McDaniel, Email
• Dr. Sharon L. Juliano, Core PI, Email

Biolistic Transfection

Location: USUHS Building C, room C2024

Equipment:

• Helios Gene-Gun® (Bio-Rad) system.
• During biolistic transfection, a plasmid DNA construct is coated onto colloidal gold particles and then introduced into cells under a brief, high-pressure pulse of helium.
• Using this system, plasmids over-expressing fluorescent proteins or driving RNAi-interference RNA hairpins can be quickly introduced into cells, tissue slice cultures, or whole specimens(exposed brain and spinal cord).
• The main utility is for live imaging or electrophysiological recording after transfection.

Scheduling

Contact Dr. Schell. The system is usually available; transfection takes less than 10 min. Users are expected to produce their own DNA-coated gold particles, using a protocol and equipment available in the Schell laboratory.

Access to the use of these cores is subject to CNRM review and chargeback fees may apply.

Points of contact for information on core resources, policies, and procedures

• Dr. Michael J. Schell, Email
• Dr. Sharon L. Juliano, Core PI, Email

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