Today is Wednesday, Dec. 13, 2017

Department of

Pharmacology & Systems Physiology 

Live Microscopy Core

Manager: Chet Closson 
Tel: (513) 558-4607
Director: Christian I. Hong, PhD 
Location: MSB 3155 

 

The Live Microscopy facility is designed to help investigators do high-resolution imaging with either living or fixed specimens. Please acknowledge use of our facility in your publications as: Live Microscopy Core, Department of Molecular and Cellular Physiology, University of Cincinnati.

Equipment available in the facility:

  • Two advanced Zeiss laser scanning microscopes:
    • LSM510 NLO confocal microscope with additional equipment for two-photon microscopy*
    • LSM 710 Live DUO confocal microscope with two parallel scanners/detectors including a very high speed scanner (>100 images/sec)*
  • Laser Capture Microdissection instrument
  • Stereomicroscope
  • Infrared imager
  • Cryostat

*These microscopes can be adapted to operate in either an inverted or upright configuration by using an objective inverter apparatus, which allows for more flexibility in sample preparations and set up. 

The Core includes an integrated animal surgery room, and maintains an IACUC approved space for holding experimental mice imported from UC and CCHMC laboratories. Investigators can use any of their mutant mice in the facility with minimal delay or paperwork. The core has environmental chambers for temperature control on the microscopes during imaging of live cells/tissues, and perfusion chambers and perfusion systems allowing continual refreshing of nutrient solutions bathing tissues. 

Training on each instrument must be completed before independent access is allowed, but then card key access allows 24/7 experimentation.


To schedule time on any instrument, please log in to your account at LabAgenda

Microscopes 

Zeiss LSM710 LIVE Duo Confocal Microscope

  • Available 24/7
  • Combination system of the new LSM710 high-resolution confocal plus the LIVE high-speed line scanner (up to 120 frames/sec at 512x512) on the same microscope stand
  • Microscope: Zeiss Axio Observer Z1 inverted microscope platform
  • Environmental chamber: encloses microscope to allow temperature and gas control
  • Microscope objectives: 10x/0.3 EC Plan Neofluar, 20x/0.8 Plan-Apochromat, 40x/1.1 LD C-Apochromat (water immersion), 63x/1.2 C-Apochromat (water-immersion)
  • LSM710 Point Scanner: 6 laser lines (405, 458, 488, 514, 560, 633nm) and 4 detectors (3 spectral confocal fluorescence plus 1 transmitted light detector).
  • LIVE fast line scanner: 3 laser lines (405, 488, 560nm) and 2 confocal fluorescence detectors
  • Fluorescence filter blocks for direct visualization of double and triple labeled samples by eye

 



Zeiss LSM510 META Confocal Microscope

  • Available all mornings and weekends by arrangement
  • Microscope: Zeiss Axiovert 200 inverted microscope platform
  • Microscope objectives: 40×/1.2 C-Apochromat (water immersion), 10×/0.45 C-Apochromat (water immersion), 20×/0.8 Plan-Apochromat (air), and 10×/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


Zeiss LSM510 NLO Two-Photon Microscope

  • Available 24/7
  • Two-photon and confocal microscope
  • Microscope: Zeiss Axiovert 200 inverted microscope platform
  • Dark environmental chamber: encloses microscope to allow 37-°C control and keep stray light out of detectors
  • Microscope objectives: 40×/1.2 C-Apochromat (water immersion), 10×/0.45 C-Apochromat (water-immersion), 40×/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


Zeiss Digital-Imaging Fluorescence Microscope

  • Available 24/7
  • Live cell ratio imaging (Ca2+and pH-sensitive dyes)
  • CoolSnap HQ camera for image acquisition


Leica Stereo Microscope

  • Available 24/7
  • Continuous zoom from 7× to 230× magnification
  • Color camera to capture images in bright field, dark field, or fluorescence modes
  • Filters for UV and GFP fluorescence excitation


Microscope Fees


Description Internal Rate External Rate
Microscopes: Independent use

Prime Hours Usage (per hour, Mon-Fri, 8:00am-5:00pm)

Off Hours Usage (per hour, all times not covered by Prime Hours)

$35

$25

Internal + Market

Internal + Market

Technician Assistance:

Full Training for New Users(per person per 4 hr minimum)

Advanced Training for Users w/ Previous Training (per person)

$280

$140

Internal + Market
Leica stereo microscope
Any-time usage fee (per hour) $5 Internal + Market
VERITAS laser-capture microdissection
(unlimited access per year) $350 Internal + Market
Li-Cor Odyssey infrared imager
(unlimited access per year) $200 Internal + Market
Applied Biosystems StepOnePlus RT-PCR
(unlimited access per year) $200 Internal + Market
Leica Cryostat
(unlimited access per year) $200 Internal + Market
PerkinElmer EnVision Multimode Plate Reader
(unlimited access per year) $200 Internal + Market
Technician Assistance:

Collect Images or samples (per hour)

Consultation (per hour)

$70

$33

Internal + Market

Internal + Market


*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
  • 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.uc.edu'. Click on the computer name "mcpbe server (Samba, Ubuntu) (mcpbe.uc.edu)" when it appears.
  • Click on the username (PI) if you are logged in your own system with the same credentials you have supplied for microscopy/server use. You will find your files in the META folder for the confocal, the NLO folder for the 2 photon, etc.
  • Otherwise supply your credentials (username and password) when the login window appears (on-campus or VPN access only).
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
  • Interactive Java tutorials about confocal microscopy at OlympusConfocal.com
  • Confocal microscopy information from Zeiss
  • Interactive database of fluorescent dye spectra at Invitrogen.com (Molecular Probes)
  • Fluorescent dye and filter database from Zeiss


Laser-Capture Microdissection


Life Technologies ArcturusXT Laser-Capture Microdissection (LCM) Instrument

  • 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 (EXFO Excite 2000-hour 1500-hour metal-halide lamp)
  • Microscope objectives: 2×, 10×, 20×, and 40× Nikon CFI60 infinity-corrected objectives
  • Motorized stage with positions for 3 standard slides or 2 large format slides, and 4 caps


Infrared Imaging



Li-Cor Odyssey Infrared Imager

  • 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

Digestive Health Center members: Please acknowledge PHS grant DK078392 in all publications involving the use of DHC cores or support.