Auditory & Vestibular Technology (AVT) Core

• Electrophysiology (Sarath Vijayakumar)
  • Contact Sarath Vijayakumar (SarathVijayakumar@creighton.edu)

• Molecular Biology (Sarath Vijayakumar)
  • Contact Sarath Vijayakumar (SarathVijayakumar@creighton.edu)

• Imaging (Anthony Stender)
  • Contact Anthony Stender (AnthonyStender@creighton.edu)

• Mass Spectrometry (Molly McDevitt)
  • Contact Molly McDevitt (MollyMcDevitt@creighton.edu)

The electrophysiology facilities are composed of the main facility on the Creighton campus (Rooms 304 and 334 in the Criss I Building) and two satellite facilities at BTNRH and UNMC.

- The electrophysiology facility offers these services:

•  Auditory Brainstem Response (ABR)
• Distortion Product Otoacoustic Emission (DPOAE)
• Wideband Acoustic Immitance (WAI). Coming soon...
• Endocochlear Potential (EP)
• Cochlear Microphonics (CM)
• Vestibular sensory Evoked Potential (VsEP)
• Video oculography for VOR/OKR

Current Instruments include:

• TDT RZ6 system for measuring auditory evoked potentials and otoacoustic emissions.
• Axopatch 200B intergrating patch clamp amplifiers and 1440A Digidata boards for recording cochlear potentials (CM, CAP, EP) from small mammals and microphonic response from zebrafish.
• National Instruments (NI) based system for vestibular evoked potentials.
• ISCAN 200 based system for VOR/OKR

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*Directed by Sarath Vijayakumar

Located in Room 209 of Criss I building, the facility is equipped with instruments commonly used for molecular biology and genomic experiments.

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Milliplex Analyzer developed by Luminex uses multiplex xMAP bead technology that allows for teh simultaneous analysis of a broad selection of analytes (e.g. cytokines, hormones and adipokines) across a wide range of disease states, including metabolic disease, inflammation, neurodegenerative disease, toxicity, cancer and more.  With the Milliplex system, you can:

• Quantitate up to 100 protein targets from culture media, sera, and other matrices simultaneously.
• Automatically analyze up to 96 samples in 60 minutes (general format: 88 sample wells + 8 standard wells)
• Dramatically increase the amount of useful data

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The QuantStudio 3 Real-Time PCR System in designed for users who need an affordable, easy-to-use real-time PCR system that doesn't compromise performance and quality.  The simplified Design and Analysis software is ideal for both first-time and experienced users.  Using Proven OptiFlex technology (featuring 4 coupled channels and white LED) and featuring three independent Veriflex temperature zones, the QuantStudio 3 system enables improved data accuracy and sensitivity for a broad range of genomic applications.  Applied Biosystems ProFlex thermocycler is also available for routine PCR applications.

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The ChemiDoc MP Imaging System is a full-feature instrument for imaging and analyzing gels and western blots.  It is designed to address multiplex fluorescent western blotting, chemiluminescence detection, general gel documentation applications, and stain-free technology imaging needs.  Features include:

• All-in-one flexible imaging - get precise, reproducible fluorescent, chemiluminescence, and colorimetric gel and blot detection, analysis and documentation in a single system
• Multiplex fluorescent western blotting - detect up to three proteins simultaneously and eliminate the need for stripping and reprobing
• Ease of use - features include automatic selection of optimal light source by application, auto focus, auto exposure, and preview features ensure optimal images.  Learn in minutes with on screen help.
• Stain-free protein normalization - stain-free imaging permits the normalization of bands to total protein in both in gels and blots; eliminates the need for housekeeping proteins
• Sample Trays and Smart Tray Technology - three sample trays available to cover diverse imaging applications. Smart Tray Technology automatically recognizes the application-specific trays and adjusts imaging parameters and software options accordingly

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The 10X Chromium Controller uses advanced microfluidics to perform single cel activity in multiple dimensions, including gene expression, cell surface proteins, immune clonotype, antigen specificity, and chromatin accessibility.  It is compatible with Chromium Singe Cell products, including Single Cell Gene Expression, Single Cell Immune Profiling, and Single Cell ATAC.

• Process 1-8 samples per run
• Collect 500-10,000 cells per sample
• Libraries produced are compatible with Illumina NextSeq 2000 in the innovative Genomics and Bioinformatics core

*Directed by Sarath Vijayakumar

We offer multiple advanced research microscopes that are available for imaging-based experiments.  We also provide a selection of sophisticated software packages that can be utilized for image analysis.  Please inquire if you have specific questions about any of these instruments or about gaining access to them.  We are happy to provide training as well as to consult with users about their experimental and analysis needs.  New users must receive official training and sign a user agreement prior to gaining access to the facility's tools or its reservation systems.

Instrumentation:

The Advanced Imaging Core hosts four confocal laser scanning microscopes from Zeiss for users who wish to perform fixed cell or live cell imaging.  All four microscopes offer DIC imaging capabilities and popular objective magnifications (10X, 20X, 40X, 63X). The two LSM 700s offer four excitation wavelengths (405, 488, 555, 639 nm).  The inverted LSM 700 also has a stage incubator.  The LSM 710 has 7 laser lines (405, 458, 488, 514, 561, 594, 633), 2 PMT’s, and a 32-channel detector.  The latest addition to the core facility is the Zeiss 980 with Airyscan, which offers high resolution, improved sensitivity, and decreased acquisition times.  Finally, the AVT- Advanced Imaging Core manages a Zeiss PALM CombiSystem, which enables users to perform microdissection and micromanipulation of samples on standard glass slides within a single microscope.

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The Advanced Imaging team also manages several specialized microscopes associated with CU-IBIF, Creighton University’s Integrated Biomedical Imaging Facility. 

First is an inverted Nikon Ti-E confocal microscope with a Yokogawa spinning disk, a Hamamatsu Orca Flash camera, and an incubated stage.  This microscope is capable of fast full-frame imaging at speeds up to 1000 frames/second, and it can perform Z-stacks at rates 10 times faster than a typical point scanning confocal.  This microscope is equipped with Live Super-Resolution, thus enabling the high-speed imaging which is ideal for live cell experiments.  It is also an ideal choice for extended time-lapse imaging experiments.

Next is an upright Leica SP8 confocal microscope that is suitable for live animal imaging and can support accessories for electrophysiology measurements.  In addition to visible-range lasers, it has a tunable, pulsed, near-infrared Ti:S laser for multiphoton excitation of UV/Visible fluorophores and second harmonic imaging. Non-descanned Super HyD detectors and a Becker and Hickl SPC 830 Time Correlated single photon counting system enable fluorescence lifetime imaging, fluorescence correlations spectroscopy, and lifetime-based FRET techniques. The Leica has spectral detectors that enable spectral imaging.

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Third is a home-built Total-Internal Reflection Fluorescence Microscope.  This instrument offers a through-the-prism beam path to reduce noise from scattered laser light, and it incorporates three objective options (10X, 20X, 40X) and four laser lines (405, 488, 552, and 647 nm).  Images are captured with an Andor iXon EMCCD scientific camera.

The team also manages an inverted ImageXpress Micro 4 widefield microscope from Molecular Devices, equipped with an incubation chamber.  The ImageXpress enables users to conduct preprogrammed experiments over extended time periods at high-throughput.  In addition, CU-IBIF is also the home to two other widefield microscopes for basic imaging needs.

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Software Packages Available:

The Advanced Imaging Core provides a selection of analysis tools for the THC and users of our facility.  We have two workstations that run the Imaris Microscopy Image Analysis software, which is helpful in processing 3D and 4D images.  A standalone workstation is available for utilizing Nikon’s NIS Elements and Leica’s LAS X software.  Users interested in knowing more about these software packages and accessing them are asked to reach out to the team.  We are happy to discuss your image processing needs with you, and we have experience in using a variety of other programs as well for analysis and processing, including ImageJ, MATLAB, Python, and R.

When publishing a research paper that relied on equipment used within the AVT Core Facility, please remember to acknowledge the core.  Proper acknowledgment is critical for tracking research papers and meets the requirements of our funding agencies.  We recommend that you use the statement below that best fits your paper (you may simply copy and paste this content into the Acknowledgments section of your paper):

If Imaging Core was used:

This research was partially conducted at the Auditory and Vestibular Technology Core (AVT) at Creighton University, Omaha, NE (RRID:SCR_023866). This facility is supported by the Creighton University School of Medicine and grants GM103427 and GM139762 from the National Institute of General Medical Science (NIGMS), a component of the National Institutes of Health (NIH). IBIF was constructed with support from grants from the National Center for Research Resources (RR016469) and the NIGMS (GM103427). This investigation is solely the responsibility of the authors and does not necessarily represent the official views of NIGMS or NIH.

If Imaging Core was not used:

This research was partially conducted at the Auditory and Vestibular Technology Core (AVT) at Creighton University, Omaha, NE. This facility is supported by the Creighton University School of Medicine and grants GM103427 and GM139762 from the National Institute of General Medical Science (NIGMS), a component of the National Institutes of Health (NIH). IBIF was constructed with support from grants from the National Center for Research Resources (RR016469) and the NIGMS (GM103427). This investigation is solely the responsibility of the authors and does not necessarily represent the official views of NIGMS or NIH.