Live Cell Imaging Facility (LCIF) of the Center for Investigation of Membrane Excitability Diseases (CIMED) is focused on studying cell signaling in excitable cells with the use of optical live cell imaging methods. We provide instruments, expertise and assistance necessary to monitor cell functions, particularly intracellular ion concentrations, with the use of fluorescent indicators. The facility is available to researchers from and outside of Washington University to carry out collaborative projects related to the mission of the Center.

Core Description

The Live Cell Imaging Center provides access to instruments that allow noninvasive observation of cellular functions, mostly intracellular free ion concentration and membrane potential changes, with the use of fluorescent indicators in living cells. After completing mandatory training, the users are granted access to the facility and can reserve time on a particular instrument using a web-based calendar. Users who choose to work independently can rely on technical assistance and advice from the staff members. Alternatively, the experiments can be performed by an LCIF staff member.  Please direct general questions and collaboration offers to LCIF Director Dr. Colin Nichols and technical inquiries to Kris Hyrc. ​

Access

Service available to all entities, including for-profit organizations.

Priority service for Washington University only.

Additional information:

Orientation and training required. Please contact Kris Hyrc for details.

Services

  • Intracellular free calcium concentration measurements in living cells with the use of ratiometric (e.g., fura-2) and/or single wavelength (e.g., fluo-3) fluorescent calcium indicators
  • Determination of intracellular free ion concentrations such as K+, Na+, Zn2+, and Mg2+ with the use of fluorescent calcium indicators in living cells
  • Measurements of cytosolic pH in living cells
  • Monitoring the changes in cell and mitochondrial membrane potential with the use of fluorescent indicators in living cells
  • Confocal imaging of fixed or living samples in FITC, TRITC and Cy5 channels

Equipment

  • Two Till Photonics digital microscopes equipped with Polychrome V monochromators, high resolution (1344x1200) cooled CCD cameras, perfusion system, and controlled environment chambers
  • A standard filter wheel based imaging rig built around a Nikon Eclipse 300 microscope with a cooled Sensicam (640x480) camera
  • A dedicated Ionoptox© rig built around a Nikon Diaphot microscope for simultaneous analysis of sarcomere length and intracellular [Ca2+] in isolated cardiac myocytes
  • Olympus FV500 confocal microscope with 488 nm, 563nm and 633nm excitation wavelengths
  • TIRF imaging system with single molecule FRET capability built around Nikon Eclipse microscope using Nikon's DV2 technology with 488 nm, 532 nm and 633 nm laser lines
  • A microelectrode setup that allows recording single cell action potentials in whole organs such as heart
  • A wide-field imaging system built around a Zeiss Axioevrt 200M microscope equipped with a Lambda DG-4 illumination system, excitation/emission filter wheels and an EM-CCD camera. The system can be used for variety of applications ranging from cytosolic free calcium measurements with a variety of single wavelength and ratiometric indicators to standard slide imaging.

Pricing

Pricing is subject to core verification

Hourly fees are charged. The rates depend on the instrument used and time of day, and range from $5 to $50/hour. See the CIMED Live Cell Imaging Facility website for details.


AFFILIATIONS
Center for Investigation of Membrane Excitability Diseases (CIMED)