LCIF

CIMED Live Cell Imaging Facility

New and Noteworthy:

$1000 microgrants are now available to offset the charges associated with use of the facility.  For more information see contacts above or see link:

1.   Overview:

The 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 inside and outside of Washington University to carry out collaborative projects related to the mission of the Center.  The facility is available to researchers interested in carrying out collaborative projects.  On January 1, 2013, we instituted a use-based repayment and grant-supported mechanisms to cover the operation costs of the facility.

2.  Equipment:

  • Two Till Photonics digital microscopes equipped with Polychrome V monochromators, high resolution (1344×1200) 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 CCD camera.
  • A dedicated IonOptix rig built around a Nikon Diaphot microscope for simultaneous analysis of sarcomere length and intracellular [Ca2+]i in isolated cardiac myocytes.
  • Olympus FV500 confocal system based on Olympus IX70 microscope with full set of Olympus lenses (4x/0.035 to 100x/1.4), which allows collecting high resolution (up to 2048×2048) images with 488 nm, 568 nm and 633 nm excitation laser lines.
  • Nikon TE2000 based TIRF system equipped with 488 nm, 531 nm and 640 nm laser lines, a dedicated Alpha TIRF (100x/1.49) lens and a fast EMCCD camera that allows single molecule FRET imaging with the use of Nikon Dual View technology.

 

3. Services:

  • Measurement of  intracellular free ion concentrations (Ca2+, K+, Na+, H+, etc.) and their dynamics with the use of single wavelength and ratiometric  fluorescent indicators in living cells.
  • Simultaneous measurements of cardiac myocyte contractility and [Ca2+]1 changes.
  • Analysis of cell structure, function and molecular interactions with the use of confocal, TIRF and FRET imaging.
  • Assistance in designing and performing experiments employing fluorescent indicators in living cells.

 

4. Staff and Contact Information:

  • Faculty Core Director:  Colin Nichols, PhD (cnichols@wustl.edu).
  • CIMED Administrator, Ms. Jayma Mikes (E-mail: jmikes@wustl.edu).
  • Core Manager:  Krzysztof (Kris) Hyrc, PhD (314-362-4876; hyrck@neuro.wustl.edu).

For all questions regarding access, training, and technical assistance please contact  Dr.  Kris Hyrc.

General inquiries should be directed to Dr. Colin Nichols or Ms. Jayma Mikes.

5. Charges:

We charge the following fees for using the facility:

Instrument

Peak Time*

Mon 9:00 AM-

Fri 5:00PM

Off-Peak Time*

Mon 5:01 PM – Fri. 9:00 PM

Nights/Weekends/Holidays*

Fri 9:01 PM –

Mon. 8:59 AM

Training fees

1-3 people

TIRF/FRET

$40/hour

$20/hour

$10/hour

$100

IonOptix

$20/hour

$10/hour

$5/hour

$50

FV500 confocal

$20/hour

$10/hour

$5/hour

$100

Till Photonics

$20/hour

$10/hour

$5/hour

$100

 *These charges apply to work done by the users themselves.  IF the work is performed by staff members, a flat fee of $50/hour applies.

 

6.  References

  1. Toib A., Zhang HX., Broekelmann TJ., Hyrc KL., Guo Q., Chen F., Remedi MS., and Nichols, CG. (2012) Cardiac specific ATP-sensitive K+ channel (KATP) overexpression results in embryonic lethality. J Mol Cell Cardiol  53, 437-445
  2. D’Avanzo N., Hyrc K., Enkvetchakul D., Covey DF., and Nichols CG. (2011) Enantioselective protein-sterol interactions mediate regulation of both prokaryotic and eukaryotic inward rectifier K+ channels by cholesterol. PLoS One  6, 1-7
  3. Zhang S., Hyrc K., Wang S., and Wice, BM.,  (2012) Xenin-25 increases cytosolic free calcium levels and acetylcholine release from a subset of myenteric neurons. Am J Physiol   303, G1347-1355
  4. Wang S., Makhina EN., Masia R., Hyrc KL., Formanack, ML., and Nichols CG. (2013) Domain organization of the ATP-sensitive potassium channel complex examined by fluorescence resonance energy transfer. J Biol Chem  288, 4378-4388
  5. Zhu T., Chappel JC., Hsu FF., Turk J., Aurora R., Hyrc K., De Camilli P., Broekelmann TJ., Mecham RP., Teitelbaum SL., and Zou, W. (2013) Type I phosphotidylinosotol 4-phosphate 5-kinase gamma regulates osteoclasts in a bifunctional manner. J Biol Chem  288, 5268-5277
  6. Hyrc K., Minta A., Escamilla PR., Chan PP., Meshik X., and Goldberg MP. (2013) Synthesis and Properties of Asante Calcium Red – a novel family of long excitation wavelength calcium indicator. Cell Calcium in print