Comparison of calcium imaging in dorsal root ganglion neurons by using laser scanning confocal and two-photon microscopy

Yimei Huang; Hongqin Yang; Jiangxu Chen; Xiuqiu Shen; Liqin Zheng; Yuhua Wang; Shusen Xie

doi:10.1117/12.908220

9 February 2012 Comparison of calcium imaging in dorsal root ganglion neurons by using laser scanning confocal and two-photon microscopy

Yimei Huang, Hongqin Yang, Jiangxu Chen, Xiuqiu Shen, Liqin Zheng, Yuhua Wang, Shusen Xie

Author Affiliations +

Proceedings Volume 8226, Multiphoton Microscopy in the Biomedical Sciences XII; 82262O (2012) https://doi.org/10.1117/12.908220
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

As one of the most important second messengers, calcium in nerve cells plays a critical role in neuronal processes, including excitability, neurotransmitter release, synaptic plasticity. Modulation of the calcium concentration is an important means of regulating diverse neuronal functions. To evaluate the role of calcium, quantitative measurement of cytosolic free calcium concentrations is necessary. There are several optical techniques that are available for measurement of calcium in live cells. Laser scanning confocal microscopy and two-photon microscopy are two prevalent techniques for their advantage in spatial resolution. In this paper, calcium in dorsal root ganglion neurons was imaged by laser scanning confocal microscopy and two-photon microscopy with Fluo-3, a calcium specific fluorescence probe. Both of spatial resolution and photobleaching, two common limitations of optical image modality, were compared between laser scanning confocal microscopy and two-photon microscopy, respectively. Three dimension images showed that laser scanning confocal microscopy and two-photon microscopy had not only similar lateral resolution but also parallel vertical resolution. However, Laser scanning confocal microscopy had an advantage over the two-photon microcopy in photobleaching. These results indicated that laser scanning confocal microscopy was more suitable than two-photon microscopy to be applied in imaging calcium in dorsal root ganglion neurons with Fluo-3.

Citation Download Citation

Yimei Huang, Hongqin Yang, Jiangxu Chen, Xiuqiu Shen, Liqin Zheng, Yuhua Wang, and Shusen Xie "Comparison of calcium imaging in dorsal root ganglion neurons by using laser scanning confocal and two-photon microscopy", Proc. SPIE 8226, Multiphoton Microscopy in the Biomedical Sciences XII, 82262O (9 February 2012); https://doi.org/10.1117/12.908220

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