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A clarifying method that improves imaging of Aplysia ganglia

CHEN Ting-Ting, YU Wei, LIU Dan-Dan, YU Ke, CHEN Song-An, WANG Ye, YANG Shao-Zhong, JIA Ruo-Nan, ZHENG Yu-Tong, HUANG Zhen, Ferdinand S Vilim, DONG Lei, Elizabeth C Cropper, Klaudiusz R Weiss, JING Jian

1State Key Laboratory of Pharmaceutical Biotechnology, Advanced Institute for Life Sciences, School of Life Sciences, Nanjing University, Nanjing 210046, China； 2Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA

Abstract

Improvements in the imaging of neural circuits are essential for studies of network function in both invertebrates and vertebrates. Therefore, CLARITY, a new imaging enhancement technique developed for mouse brains has attracted broad interest from researchers working on other species. We studied the potential of a modified version of CLARITY to enhance the imaging of ganglia in an invertebrate Aplysia. For example, we have modified the hydrogel solution and designed a small container for the Aplysia ganglia. The ganglia were first processed for immunohistochemistry, and then for CLARITY. We examined the compatibility of these techniques and the extent to which the imaging of fluorescence improved using confocal microscopy. We found that CLARITY did indeed enhance the imaging of CP2 immunopositive neurons in Aplysia ganglia. For example, it improved visualization of small, weak immunoreactive neurons deep in the ganglia. Our modifications of CLARITY make this new method suitable for future use in Aplysia experiments. Furthermore, our techniques are likely to facilitate imaging in other invertebrate ganglia.

Key words: CLARITY; imaging enhancement ; neuronal structure ; Aplysia ; immunohistochemistry

Received: 2017-01-22　　Accepted: 2017-02-22

Corresponding author: 景键　　E-mail: jingj01@nju.edu.cn

Citing This Article：

CHEN Ting-Ting, YU Wei, LIU Dan-Dan, YU Ke, CHEN Song-An, WANG Ye, YANG Shao-Zhong, JIA Ruo-Nan, ZHENG Yu-Tong, HUANG Zhen, Ferdinand S Vilim, DONG Lei, Elizabeth C Cropper, Klaudiusz R Weiss, JING Jian. A clarifying method that improves imaging of Aplysia ganglia. Acta Physiol Sin 2017; 69 (4): 461-466