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Calcium Imaging

The Calcium Imaging Research Support Laboratory 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

The Nikon High-Speed Multidimensional Image Analysis System is a wide-field system specifically designed for visualizing live cells. Our system includes a high-speed CCD camera, filter wheels and shutter, x-y motorized stage and z-focus control. Filter sets are available for all commonly used fluorescent dyes, including genetically encoded indicators. We also have a variety of stage-mounted incubators for long term imaging of cells or tissues under culture conditions. The system is run by the latest version of MetaMorph Microscopy Automation and Image Analysis Software, which provides fully programable control of image acquisition as well as stage and focus positioning.

Confocal Microscopy

The Visitech VT-Infinity 2D Confocal provides high temporal and spatial multicolor imaging. The system is equipped with stage incubators and perfusion baths, as well as 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. A krypton argon (488, 568 and 647 nm) and a photodiode (405 nm) laser 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 also runs MetaMorph software for full control of image capture and data analysis.

Plate Reader

The 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.

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

Recent Publications

  • Candela, J., R. Wang, and C. White, Microvascular Endothelial Dysfunction in Obesity Is Driven by Macrophage-Dependent Hydrogen Sulfide Depletion. Arterioscler Thromb Vasc Biol, 2017. 37(5): p. 889-899.

  • Wang, J., et al., Role of cystathionine-gamma-lyase in hypoxia-induced changes in TASK activity, intracellular [Ca2+] and ventilation in mice. Respir Physiol Neurobiol, 2017. 246: p. 98-106.

  • Zhao, G., et al., N-(3-oxo-acyl) homoserine lactone inhibits tumor growth independent of Bcl-2 proteins. Oncotarget, 2016. 7(5): p. 5924-42.

  • Demczuk, M., et al., Retinoic Acid Regulates Calcium Signaling to Promote Mouse Ovarian Granulosa Cell Proliferation. Biol Reprod, 2016. 95(3): p. 70.

  • White, C., A. Nixon, and N.A. Bradbury, Determining Membrane Protein Topology Using Fluorescence Protease Protection (FPP). J Vis Exp, 2015(98).

  • Velmurugan, G.V., et al., Depletion of H2S during obesity enhances store-operated Ca2+ entry in adipose tissue macrophages to increase cytokine production. Science Signaling, 2015. 8(407): p. ra128.

  • Rajan, S., et al., Structural transition in Bcl-xL and its potential association with mitochondrial calcium ion transport. Scientific Reports, 2015. 5: p. 10609.

  • Kim, D., et al., Hydrogen sulfide and hypoxia-induced changes in TASK (K2P3/9) activity and intracellular Ca2+ concentration in rat carotid body glomus cells. Respir Physiol Neurobiol, 2015. 215: p. 30-8.