Calcium Imaging

 


The Calcium Imaging Research Support Laboratory, housed in the Department of Physiology and Biophysics, provides the resources and expertise necessary to carry out state-of-the-art intracellular calcium measurements in cultured or freshly isolated single cells. Three instruments are available: a widefield fluorescence microscope suitable for timecourse calcium monitoring in individual cells, a multi-point confocal system for high spatial and temporal resolution measurements, and a plate reader for multi-well format recordings. To initiate a new project please make arrangements with the Lab Director to discuss feasibility and technical approach.

Equipment and Capabilities

Widefield Fluorescence Microscopy

Built around an inverted Olympus IX-71 microscope the system is equipped with a Hamamatsu CCD, motorized stage, and heated perfusion chambers with rapid solution switching. A Lambda DG4 is used for rapid switching of the excitation wavelength and a Lambda 10-3 for rapid switching of emission wavelength. Filter sets are available for all commonly used fluorescent dyes, including genetically-encoded indicators. The microscope is controlled by Compix Simple PCI software for image capture and data analysis.

Confocal Microscopy

The Visitech VT-Infinity 2D Confocal provides high temporal and spatial multicolor imaging. The scan-head is mounted on an Olympus IX-71 microscope equipped with a Hamamatsu EM-BT CCD camera, stage incubator and perfusion bath, and a motorized stage and z-axis. The VT-Infinity is a multi-point confocal system that provides fast scan rates and exceptionally low photo-bleaching. The system is therefore ideally suited for live cell experiments requiring extended period, high frame rate acquisition. Krypton argon (488, 568 and 647 nm) and a photodiode (405 nm) laser with acoustic optic couplers provide the excitation, and dual view MicroImager Emission Bean Splitters are available for use with dual emission dyes and fluorescence resonance energy transfer (FRET) studies. The system runs Hamamatsu HCImage software for image capture and data analysis.

BMG Polar Star Optima plate reader

Performs real-time kinetic monitoring of fluorescence, luminescence, or absorbance in cells grown in 24 well, 96 well or 384 well plates. Additional features include: two programmable reagent injectors, temperature control up to 60°C, and analysis software features for endpoint, long-term and fast kinetic measurements. Also available are an Eppendorf epMotion model 5070 Liquid Handling Workstation and a Tomtec Quadra 96 with indexing unit for studies using the 96 and 384 well formats.

Fee Schedule

 Widefield system:

 RFUMS users $25 ($40 outside users) per half day (0 – 4 hours)

 Confocal system:

 RFUMS users $25 ($40 outside users) per half day (0 – 4 hours)

 Plate Reader:

 RFUMS users $15 ($25 outside users) per hour

Contacts


Director:
  Dr. Carl White
carl.white@rosalindfranklin.edu
Tel: 847-578-8358

Research Specialist: Emily Eckenrode
emily.eckenrode@rosalindfranklin.edu

Tel: ext 7816

Recent Publications

  • Eckenrode, E.F., Yang, J., Velmurugan,G.V., Foskett, J.K. and White, C. (2010) Journal of Biological Chemistry. 285(18), 13678-13684.
  • Paul, A.G., Sharma-Walia, N., Kerur, N., White, C. & Chandran, B. (2010) Cancer Research. 70(9), 3697- 3708.
  • Wang, X., Olberding, K.E., White, C. and Li, C. (2011) Cell Death & Differentiation. 18, 38-47.
  • Velmurugan, G.V. and White, C. (2012) American Journal of Physiology (Heart Circ Physiol), 302, H124-H134.
Life in Discovery
Calcium Imaging Links
3333 Green Bay Road, North Chicago, Il 60064-3095 • 847-578-3000