Share Email Print
cover

Proceedings Paper

Visualizing substructure of Ca2+ waves by total internal reflection fluorescence microscopy
Author(s): Yongqiang Bai; Aihui Tang; Shiqiang Wang; Xing Zhu
Format Member Price Non-Member Price
PDF $14.40 $18.00
cover GOOD NEWS! Your organization subscribes to the SPIE Digital Library. You may be able to download this paper for free. Check Access

Paper Abstract

Total internal reflection fluorescence microscope is a new optical microscopic system based on near-field optical theory. Its character of illumination by evanescent wave, together with the great signal-to-noise ratio and temporal resolution achieved by high quality CCD, allows us to analyze the spatiotemporal details of local Ca2+ dynamics within the nanoscale microdomain surrounding different Ca2+ channels. We have recently constructed a versatile objective TIRFM equipped with a high numerical aperture (NA=1.45) objective. Using fluo-4 as the Ca2+ indicator, we visualized the near-membrane profiles of Ca2+ waves and elementary Ca2+ sparks generated by Ca2+ release channels in rat ventricular myocytes. Different from those detected using conventional and confocal microscopy, Ca2+ waves observed with TIRFM exhibited fine inhomogenous substructures composed of fluctuating Ca2+ sparks. The anfractuous routes of spark recruitment suggested that the propagation of Ca2+ waves is much more complicated than previously imagined. We believe that TIRFM will provide a unique tool for dissecting the microscopic mechanisms of intracellular Ca2+ signaling.

Paper Details

Date Published: 9 February 2005
PDF: 8 pages
Proc. SPIE 5635, Nanophotonics, Nanostructure, and Nanometrology, (9 February 2005); doi: 10.1117/12.580734
Show Author Affiliations
Yongqiang Bai, Peking Univ. (China)
Aihui Tang, Peking Univ. (China)
Shiqiang Wang, Peking Univ. (China)
Xing Zhu, Peking Univ. (China)


Published in SPIE Proceedings Vol. 5635:
Nanophotonics, Nanostructure, and Nanometrology
Xing Zhu; Stephen Y. Chou; Yasuhiko Arakawa, Editor(s)

© SPIE. Terms of Use
Back to Top