The Integrated Biomedical Imaging Facility at Creighton University is a new multi-user core that is closely related to the scientific focus in structural biology. Core technology includes the LSM 510 METANLO confocal scanning system with three lasers (Argon 458/477/488/514 nm, Green HeNe 543 nm, Red HeNe 633 nm. The confocal is built on a Zeiss AxioPlan 2IE MOT motorized upright microscope, with the following objectives: Plan Neo 40x oil 1.3 numerical aperture (n.a.) with differential interference contrast (DIC); Plan 63x 1.4 n.a. DIC; Achroplan 40x 0.8 n.a. water DIC, for live cell work; C-Apochromat 40x 1.2 n.a. DIC ,for fluorescence correlation spectroscopy; Plan Apo 10x; Plan Neo 20x 0.75 n.a. DIC. Initial funding for the core was through an National Science Foundation EPSCoR grant. The Core can image with most fluorophores, in either single-photon or multi-photon modes, with resolution approaching the theoretical limit (222 nm) The META multi-channel system enables separation of fluorophores with closely-spaced emission peaks by linear un-mixing. The types of experiments available far surpass those of conventional high resolution confocal imaging. Particular emphasis is placed on fluorescence-based and other dynamic imaging modalities, including:
- Spectral Imaging using the META detector, including linear un-mixing for separation of fluorophores with overlapping emission bands.
- Fluorescence Correlation Spectroscopy (FCS) for the study of the dynamics of molecular interactions.
- Fluorescence Recovery After Photobleaching (FRAP), for the study of molecular mobility.
- Fluorescence Resonance Energy Transfer (FRET), for the study of molecular interactions.
- Fluorescence Lifetime Imaging (FLIM), an enhanced approach to FRET.
- Total Internal Reflection Fluorescence (TIRF), for the study of single molecule dynamics.
- Complex time-series experiments from seconds to days.
- Imaging of ionic currents of pH.
- Live cell imaging in culture with temperature control.
- Metabolic imaging.
- Deep brain or deep tissue imaging in vitro or in vivo.
Director CUIBIF Integrated Biomedical Imaging Facility
Department of Biomedical Sciences
2500 California Plaza
Omaha, NE 68178