Comparison of seven optical clearing methods for mouse brain

Peng Wan; Jingtan Zhu; Tingting Yu; Dan Zhu

doi:10.1117/12.2289392

12 February 2018 Comparison of seven optical clearing methods for mouse brain

Peng Wan, Jingtan Zhu, Tingting Yu, Dan Zhu

Proceedings Volume 10481, Neural Imaging and Sensing 2018; 104811I (2018) https://doi.org/10.1117/12.2289392
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

Recently, a variety of tissue optical clearing techniques have been developed to reduce light scattering for imaging deeper and three-dimensional reconstruction of tissue structures. Combined with optical imaging techniques and diverse labeling methods, these clearing methods have significantly promoted the development of neuroscience. However, most of the protocols were proposed aiming for specific tissue type. Though there are some comparison results, the clearing methods covered are limited and the evaluation indices are lack of uniformity, which made it difficult to select a best-fit protocol for clearing in practical applications. Hence, it is necessary to systematically assess and compare these clearing methods. In this work, we evaluated the performance of seven typical clearing methods, including 3DISCO, uDISCO, SeeDB, ScaleS, Clear^T2, CUBIC and PACT, on mouse brain samples. First, we compared the clearing capability on both brain slices and whole-brains by observing brain transparency. Further, we evaluated the fluorescence preservation and the increase of imaging depth. The results showed that 3DISCO, uDISCO and PACT posed excellent clearing capability on mouse brains, ScaleS and SeeDB rendered moderate transparency, while Clear^T2 was the worst. Among those methods, ScaleS was the best on fluorescence preservation, and PACT achieved the highest increase of imaging depth. This study is expected to provide important reference for users in choosing most suitable brain optical clearing method.

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Peng Wan, Jingtan Zhu, Tingting Yu, and Dan Zhu "Comparison of seven optical clearing methods for mouse brain ", Proc. SPIE 10481, Neural Imaging and Sensing 2018, 104811I (12 February 2018); https://doi.org/10.1117/12.2289392

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KEYWORDS

Brain

Optical clearing

Neuroimaging

Green fluorescent protein

Tissue optics

Tissues

Transparency

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