Microscopy Core

Center for Neuroscience and Regenerative Medicine

Microscopy Core

Microscopy Core

The major functions of the Microscopy Core are as follows:

  • Provide specialized fluorescence and bright field microscopes for quantitative and qualitative analysis of molecular processes at the resolution of single cells as well as across circuits of neurons.
  • Operate advanced systems for two-photon microscopy integrated with electrophysiology and for quantitative unbiased stereology. The fluorescence microscopy is well designed to exploit the advantages of pre-clinical models for mechanistic approaches using optogenetics, in vivo imaging, and optical imaging through cleared tissues for whole brain three-dimensional analyses. 

 

 

 Photon Microscope/Electrophysiology

Location: USUHS B2063

     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 2012" software. Filter cube combinations support detection of green (GFP, Calcium Green, Alexa 488, etc.), red (e.g. Rhodamine dyes) and blue (DAPI, cascade blue) fluorescence.

  • Electrophysiology - Molecular Probes Axon CNS MultiClamp 700B two-channel amplifier and "pClamp 10" software for pulse generation and data acquisition, GRASS S88K Dual output stimulator and a PMP 102 micropipette puller (MDI).
  • Optogenetics – X-Cite XLED 1 light source with excitation filters for blue (470nm, channelrhodopsin)  and yellow (570 nm, Halorhodopsin), light guides and quartz fibers for optical control of rhodopsin-expressing cells in physiological experiments.

  • 3-D tissue imaging – extra-long working distance objectives  (20x) corrected for Scale or FocusClear solutions, motorized microscope stage, tiling software and off-line computer workstation with 3-D analysis software (Amira/Arivis) for 3-D analysis of large image stacks.

  • Additional equipment includes a pico-injector for bolus loading of dyes or focal stimulation, an 8-channel peristaltic pump for bath perfusion and anesthesia system for in vivo imaging.

  • Best suited for imaging slices or in vivo preparations with concurrent electrophysiological measurements.

     Scheduling:

     Contact Fritz Lischka for initial consultation and training. Use the BIC on-line scheduling system to reserve time (choose "7MP").

     Points of Contact:

 

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 to reserve time. See user guide for instructions. Contact Dennis McDaniel for initial training.

     Points of Contact:

     Dr. Dennis McDaniel, dennis.mcdaniel@usuhs.edu

     Dr. Sharon L. Juliano, Core PI

 

Biolistic Transfection

Location: Mobile

     Equipment:

  • Helios Gene-Gun® (Bio-Rad) system(link is external).
  • 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. Lischka. The system is usually available Transfection takes less than 10 minutes. Users are expected to produce their own DNA-coated gold particles.

     Points of Contact:

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

  • Access to CNRM core resources is available for CNRM funded studies and for broader research needs according to CNRM policies, including prioritization and cost sharing.  For further information on the policies or for a copy, please email julie.wilberding.ctr@usuhs.ed.

Additional systems for imaging are available through the USU Biomedical Instrumentation Center.