The Creighton University Integrated Biomedical Imaging core facility features a multi-photon confocal microscope that allows high resolution imaging of labeled cells and tissues in three dimensions and time series. The facility, available to investigators on a fee-for-use-time basis, is supported by the Creighton University School of Medicine, the Nebraska NIH-INBRE program, and by user fees.
The facility, located in Criss II room 405, has been instrumental in the creation of forums for scientific interaction among Nebraska scientists, including an annual symposium on imaging methods.
The instruments and imaging techniques now available to IBIF users include the following:
- Confocal imaging of fluorescent molecular labels in tissues and cells, using a Leica TCS SP8, with five laser lines and high numerical aperture objectives,
- Multi-photon confocal imaging using the Leica TCS SP8 confocal microscope, Spectra Physics Mai Tai DeepSee near-infrared tunable laser and Leica HyD advanced detectors,
- Long-working distance multi-photon optimized objectives for deep tissue imaging, e.g. in brain,
- Adjustable height fixed stage for live animal studies,
- Imaging of molecular dynamic processes in living cells,
- Imaging of transmembrane currents,
- Fluorescence Resonance Energy Transfer (FRET), a method of quantifying intermolecular associations,
- FLIM, which gives information about the excited state of molecules,
- Metabolic imaging, which provides a direct reading of energy utilization in living cells,
- TIRF imaging, using an Olympus TIRF system on an inverted Olympus microscope, which enables very low background visualization of membrane-bound molecules, and
- Single molecule imaging, using a very-high quantum efficiency EM-CCD camera (Andor DU-897E) on an Olympus inverted fluorescence microscope.
In addition the core is equipped with:
- Three-dimensional reconstruction and analysis software (PerkinElmer Volocity) in a stand-alone workstation,
- Open-source OMERO server for image management, and
- High-resolution transmitted light (Normaski) and epifluorescence imaging on a Zeiss Axioskop II with high numerical objectives and image acquisition via a Q-Imaging EXi camera.
Imaging can be performed with a wide variety of fluorophores, including fluorescent proteins, in either single-photon or multi-photon modes. Software is available for FRET, FRAP and FLIM, ratiometric imaging, and complex time series experiments.
Coming soon – superresolution!
Richard Hallworth, Ph.D., Director, firstname.lastname@example.org