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Live Microscopy Core
Manager: Chet Closson
Director: Marshall H Montrose, PhD
Location: MSB 4105
The Live Microscopy Core imaging facility offers two advanced laser-scanning microscopes optimized for work with living cells and tissues, plus various conventional fluorescence microscopes and a fluorescence stereomicroscope. Confocal and two-photon imaging can be performed in an inverted or upright configuration to best fit the experimental set-up. Also available for use is a Laser Capture Microdissection instrument to isolate single cell types for DNA, RNA, or proteomic analysis.
The facility is self-serve and open 24/7 to users who have undergone full training so that they can work independently. Data is automatically backed up to a remote server that allows users password-controlled access to archived data. If you prefer not to invest in full user training, Core staff can perform imaging for you for an extra fee.
The Core has capabilities for and experience with a variety of experimental techniques, such as Förster resonance energy transfer (FRET), fluorescence recovery after photobleaching (FRAP), ratio-imaging with pH- and calcium-sensitive dyes, second harmonic generation (SHG) imaging, and in vivo two-photon photo-damage models.
Microscopes
 Zeiss LSM510 META Confocal Microscope
Go to Microscope Scheduler (users only)
- Available all mornings and weekends by arrangement
- Microscope: Zeiss Axiovert 200 inverted microscope platform
- Microscope objectives: 40x/1.2 C-Apochromat (water immersion), 10x/0.45 C-Apochromat (water immersion), 20x/0.8 Plan-Apochromat (air), and 10x/0.5 Fluar (air)
- Two conventional confocal detectors
- META detector: that allows spectral analysis of emissions over the range of 460-800 nm with roughly 10-nm resolution
- Transmitted light detector for laser-light Nomarski (differential interference contrast) imaging
- Fluorescence filter blocks for direct visualization of double and triple labeled samples by eye
- View diagram of the Zeiss META confocal light-path configuration
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 Zeiss LSM510 NLO Two-Photon Microscope
Go to Microscope Scheduler (users only)
- Available 24/7
- Two-photon and confocal microscope
- Microscope: Zeiss Axiovert 200 inverted microscope platform
- Dark environmental chamber: encloses microscope to allow 37 ĄC control and keep stray light out of detectors
- Microscope objectives: 40x/1.2 C-Apochromat (water immersion), 10x/0.45 C-Apochromat (water-immersion), 40x/0.8 C-Achroplan NIR (water-immersion), and 10x/0.3 Plan-Neofluar (air)
- Lasers for excitation of fluorescence: Ar (458, 477, 488, and 514 nm), green HeNe (543nm) and red HeNe (633 nm), Ti-Sa (700-990 nm)
- Two non-descanned detectors, which allow for collection of light from deep tissue two-photon imaging with higher efficiency than confocal detectors
- Transmitted light detector for laser-light Nomarski (differential interference contrast) imaging
- Fluorescence filter blocks for direct visualization of double and triple labeled samples by eye
- View diagram of the Zeiss NLO descanned detector light-path configuration
- View diagram of the Zeiss NLO nondescanned detector light-path configuration
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Zeiss Digital-Imaging Fluorescence Microscope
- Available 24/7
- Live cell ratio imaging (Ca2+ and pH-sensitive dyes)
- CoolSnap HQ camera for image acquisition
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Leica Stereo Microscope
- Available 24/7
- Continuous zoom from 7x to 230x magnification
- Color camera to capture images in bright field, dark field, or fluorescence modes
- Filters for UV and GFP fluorescence excitation
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Microscope Fees
| Service |
Standard Rates |
Member Rates* |
| User training (for novices) |
$250/person |
Two free user trainings per PI per year, then $250/person |
| User training (for novices) |
$250/person |
Two free user trainings per PI per year, then $250/person |
| Two-photon training (and "entrance exam" for users familiar with the Zeiss LSM) |
$100/person |
$100/person |
| Primetime usage fee (Mon-Fri 9 am-5 pm) |
$30/hour |
$20/hour* |
| Offpeak usage fee (nights and weekends) |
$20/hour |
$10/hour* |
*Digestive Health Center members: First-time DHC member PIs receive 20 hours free. DHC member discounted hourly rates apply to the first 100 hours per quarter, and standard rates apply thereafter. Please acknowledge PHS grant DK078392 in all publications involving the use of DHC cores or support.
CD-R or CD-RW are provided at cost to users to facilitate data storage.
Resources
Software
- Metamorph - Universal imaging software for post-acquisition data analysis, available for sharing on computers in MSB 4103
- ImageJ - Java-based image process and analysis, free download
- PC Scion Image - Imaging software, free download
- Zeiss LSM Browser - View LSM images and acquisition information, free download
Access Microscope Backup Data Server (users only)
- With a windows 2000/XP machine, search for computer 'mcpbe' or 'mcpbe.uc.edu'. Click on the computer name "Samba 3.0.13-1.6-SUSE (mcpbe)" when it appears.
- Click on your username if you have logged in with the same credentials you have supplied for microscope use.
- Otherwise supply your credentials (username and password) when the login window appears (VPN required outside UC).
Sample preparation
- In-house mouse surgery room available
- Incubators for cell culture
- Perfusion pumps and custom chambers
Upright and Inverted Confocal Microscopes
- Objective Inverter can be applied to both the META confocal and the two-photon microscopes to convert the inverted platform into an upright configuration
Helpful Microscopy Resources
Laser-Capture Microdissection
 Molecular Devices VERITAS Laser-Capture Microdissection (LCM) Instrument
Go to LCM Scheduler (users only)
- Available 24/7
- Automated microdissection of individual cells or multicellular structures from slides containing tissue sections or cytological samples
- Harvest single cells or tens of thousands of cells in seconds
- IR laser capture and UV laser cutting
- Epifluorescence (1500-hour metal halide lamp)
- Microscope objectives: 2x, 10x, 20x, and 60x Nikon infinity-corrected objectives
- Motorized stage with positions for 3 slides and 8 caps
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Infrared Imaging
Li-Cor Odyssey Infrared Imager
Go to Li-Cor Odyssey Scheduler (users only)
- Available 24/7
- IR fluorescence detection and analysis of Western blot bands
- Two IR lasers (680nm and 780nm) for simultaneous two-color excitation and fluorescence detection
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Digestive Health Center members: Please acknowledge PHS grant DK078392 in all publications involving the use of DHC cores or support.
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