CUBIC-plus: An optimized method for rapid tissue clearing and decolorization

Zhengyu Ren; Yuzheng Wu; Zhenning Wang; Yang Hu; Jie Lu; Jinhui Liu; Yalan Chen; Maojin Yao

doi:10.1016/j.bbrc.2021.06.075

CUBIC-plus: An optimized method for rapid tissue clearing and decolorization

Biochem Biophys Res Commun. 2021 Sep 3:568:116-123. doi: 10.1016/j.bbrc.2021.06.075. Epub 2021 Jun 30.

Authors

Zhengyu Ren¹, Yuzheng Wu², Zhenning Wang³, Yang Hu², Jie Lu³, Jinhui Liu⁴, Yalan Chen⁵, Maojin Yao⁶

Affiliations

¹ School of Pharmaceutical Science, University of South China, Hengyang, 421001, China; Guangzhou Institute of Respiratory Disease & China State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510182, China.
² Guangzhou Institute of Respiratory Disease & China State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510182, China.
³ Guangzhou Institute of Respiratory Disease & China State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510182, China; Department of Neurosurgery, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer, Guangzhou, 510182, China; Department of Neuro-oncology, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer, Guangzhou, 510182, China.
⁴ The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510182, China.
⁵ School of Pharmaceutical Science, University of South China, Hengyang, 421001, China.
⁶ Guangzhou Institute of Respiratory Disease & China State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510182, China. Electronic address: maojin.yao@aliyun.com.

PMID: 34217010
DOI: 10.1016/j.bbrc.2021.06.075

Abstract

Advanced CUBIC clearing method can clear diverse organs and even the entire body of a mouse to enable high-resolution 3D imaging. Advanced CUBIC reagent has often been used due to its low toxicity and easy preparation. However, Advanced CUBIC has a long experimental cycle, which reduces the efficiency of data acquisition. In this study, we first tracked the clarity changes of different organs cleared by Advanced CUBIC and identified the shortest time required for optimal transparency in individual organs. We then introduced ultrasound processing and developed a decolorization cocktail to optimize the clearing efficiency of the Advanced CUBIC method. With the optimized clearing CUBIC-Plus method, we achieved high resolution 3D imaging of mouse organs. Our CUBIC-Plus provides an efficient procedure to clear different organ for 3D imaging at high resolution.

Keywords: 3D imaging; CUBIC; Decolorization; Optimization; Tissue clearing; Ultrasound.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Brain Neoplasms / pathology
Brain Neoplasms / secondary
Color
Female
Fluorescent Antibody Technique / methods*
Imaging, Three-Dimensional / methods*
Lung Neoplasms / pathology
Mice
Mice, Inbred C57BL
Microscopy, Confocal / methods*
Microscopy, Fluorescence / methods
Ultrasonic Waves