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[The application of confocal simultaneous scanner unit in the study of forebrain neurodevelopment in zebrafish.] [Article in Chinese]

SHI Ying, GAO Jing-Xia, PENG Gang, JIANG Min^*

Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai 200032, China

Abstract

With the application of the photoconversion technology of genetically expressed fluorescent proteins in biologic field, more powerful confocal imaging ability was demanded. The aim of the present study was to establish an experimental model employing confocal simultaneous scanner unit for simultaneous laser stimulation and imaging, taking study of forebrain neurodevelopment in zebrafish as an example. In the present study, 36-48-hour-old Tg(lhx5:kaede) zebrafish embryos were mounted with 1.2% low melting temperature agarose. The forebrain neurons marked with kaede were observed using the simultaneous scanner unit of confocal microscopy. The 405 nm laser was used to stimulate the region of interest (ROI), while 488 and 559 nm lasers were used to acquire images at the same time. The photoconversion state of kaede protein was then reviewed, and the projecting pattern of neurons stimulated by the ultraviolet laser was examined. The results showed that, the fluorescence of stimulated kaede turned from green to red, and the photoconversion of kaede demonstrated anterior dorsal telencephalon (ADt) neurons projected axons ventrally into the anterior commissure (AC) and supraoptic tract (SOT). These results suggest the confocal simultaneous scanner unit meets the demand of the photoconversion experiment. The application of confocal simultaneous scanning technology in examining Tg(lhx5:kaede) zebrafish embryos affords an ideal experimental model in neurodevelopment study.

Key words: confocal simultaneous scanner unit; zebrafish; photoconversion; kaede; neurodevelopment

Received: 2012-06-12　　Accepted: 2012-08-27

Corresponding author: 姜民　　E-mail: minjiang@fudan.edu.cn

Citing This Article：

SHI Ying, GAO Jing-Xia, PENG Gang, JIANG Min. [The application of confocal simultaneous scanner unit in the study of forebrain neurodevelopment in zebrafish.] [Article in Chinese]. Acta Physiol Sin 2013; 65 (1): 77-82 (in Chinese with English abstract).