Special Section on Light Microscopy of Connectivity

Fluorescence micro-optical sectioning tomography using acousto-optical deflector-based confocal scheme

[+] Author Affiliations
Xiaoli Qi, Tao Yang, Longhui Li, Jiancun Wang, Shaoqun Zeng, Xiaohua Lv

Britton Chance Center for Biomedical Photonics, Huazhong University of Science and Technology, Wuhan National Laboratory for Optoelectronics, 1037 Luoyu Road, Wuhan 430074, China

Huazhong University of Science and Technology, Department of Biomedical Engineering, Key Laboratory of Biomedical Photonics of Ministry of Education, 1037 Luoyu Road, Wuhan 430074, China

Neurophoton. 2(4), 041406 (Oct 15, 2015). doi:10.1117/1.NPh.2.4.041406
History: Received April 25, 2015; Accepted September 9, 2015
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Abstract.  Fluorescent labeling has opened up the possibility of clarifying the complex distribution and circuit wiring of specific neural circuits for particular functions. To acquire the brain-wide fluorescently labeled neural wiring, we have previously developed the fluorescence micro-optical sectioning tomography imaging system. This employs simultaneous mechanical sectioning and confocal imaging of the slices, and is capable of acquiring the image dataset of a centimeter-sized whole-mouse brain at a voxel resolution of 1μm. We analyze the key optical considerations for the use of an acousto-optical deflector (AOD) scanner-based confocal detection scheme in this system. As a result, the influence of confocal detection, the imaging site during sectioning, and AOD fast scan mode on signal-to-background noise ratio are described. It is shown that mechanical sectioning to separate the slice and optical sectioning by confocal detection should be combined to maximize background suppression in simultaneous fast scan imaging while sectioning system setup.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Xiaoli Qi ; Tao Yang ; Longhui Li ; Jiancun Wang ; Shaoqun Zeng, et al.
"Fluorescence micro-optical sectioning tomography using acousto-optical deflector-based confocal scheme", Neurophoton. 2(4), 041406 (Oct 15, 2015). ; http://dx.doi.org/10.1117/1.NPh.2.4.041406


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