FDISCO: Advanced Solvent-based Clearing Method for Imaging Whole Organs

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2019 Feb 13

PMID 30746463

Citations 114

Authors

Yisong Qi

Tingting Yu

Jianyi Xu

Peng Wan

Yilin Ma

Jingtan Zhu

Yusha Li

Hui Gong

Qingming Luo

Dan Zhu

Affiliations

Soon will be listed here.

Abstract

Various optical clearing methods have emerged as powerful tools for deep biological imaging. Organic solvent-based clearing methods, such as three-dimensional imaging of solvent-cleared organs (3DISCO), present the advantages of high clearing efficiency and size reduction for panoptic imaging of large samples such as whole organs and even whole bodies. However, 3DISCO results in a rapid quenching of endogenous fluorescence, which has impeded its application. Here, we propose an advanced method named FDISCO to overcome this limitation. FDISCO can effectively preserve the fluorescence of various fluorescent probes and can achieve a long storage time of months while retaining potent clearing capability. We used FDISCO for high-resolution imaging and reconstruction of neuronal and vascular networks. Moreover, FDISCO is compatible with labeling by multiple viruses and enables fine visualization of neurons with weak fluorescence labeling in the whole brain. FDISCO represents an effective alternative to the three-dimensional mapping of whole organs and can be extensively used in biomedical studies.

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References

Erturk A, Becker K, Jahrling N, Mauch C, Hojer C, Egen J . Three-dimensional imaging of solvent-cleared organs using 3DISCO. Nat Protoc. 2012; 7(11):1983-95. DOI: 10.1038/nprot.2012.119. View

Susaki E, Ueda H . Whole-body and Whole-Organ Clearing and Imaging Techniques with Single-Cell Resolution: Toward Organism-Level Systems Biology in Mammals. Cell Chem Biol. 2016; 23(1):137-157. DOI: 10.1016/j.chembiol.2015.11.009. View

Yu T, Qi Y, Gong H, Luo Q, Zhu D . Optical clearing for multiscale biological tissues. J Biophotonics. 2017; 11(2). DOI: 10.1002/jbio.201700187. View

Kasthuri N, Hayworth K, Berger D, Schalek R, Conchello J, Knowles-Barley S . Saturated Reconstruction of a Volume of Neocortex. Cell. 2015; 162(3):648-61. DOI: 10.1016/j.cell.2015.06.054. View

Saeed I, Ashraf S . Denaturation studies reveal significant differences between GFP and blue fluorescent protein. Int J Biol Macromol. 2009; 45(3):236-41. DOI: 10.1016/j.ijbiomac.2009.05.010. View

Chung K, Wallace J, Kim S, Kalyanasundaram S, Andalman A, Davidson T . Structural and molecular interrogation of intact biological systems. Nature. 2013; 497(7449):332-7. PMC: 4092167. DOI: 10.1038/nature12107. View

Tainaka K, Kuno A, Kubota S, Murakami T, Ueda H . Chemical Principles in Tissue Clearing and Staining Protocols for Whole-Body Cell Profiling. Annu Rev Cell Dev Biol. 2016; 32:713-741. DOI: 10.1146/annurev-cellbio-111315-125001. View

Silvestri L, Costantini I, Sacconi L, Pavone F . Clearing of fixed tissue: a review from a microscopist's perspective. J Biomed Opt. 2016; 21(8):081205. DOI: 10.1117/1.JBO.21.8.081205. View

Susaki E, Tainaka K, Perrin D, Kishino F, Tawara T, Watanabe T . Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis. Cell. 2014; 157(3):726-39. DOI: 10.1016/j.cell.2014.03.042. View

10.

Tainaka K, Kubota S, Suyama T, Susaki E, Perrin D, Ukai-Tadenuma M . Whole-body imaging with single-cell resolution by tissue decolorization. Cell. 2014; 159(4):911-24. DOI: 10.1016/j.cell.2014.10.034. View

11.

Lai H, Ng W, Gentleman S, Wu W . Chemical Probes for Visualizing Intact Animal and Human Brain Tissue. Cell Chem Biol. 2017; 24(6):659-672. DOI: 10.1016/j.chembiol.2017.05.015. View

12.

Tsien R . The green fluorescent protein. Annu Rev Biochem. 1998; 67:509-44. DOI: 10.1146/annurev.biochem.67.1.509. View

13.

Gong H, Xu D, Yuan J, Li X, Guo C, Peng J . High-throughput dual-colour precision imaging for brain-wide connectome with cytoarchitectonic landmarks at the cellular level. Nat Commun. 2016; 7:12142. PMC: 4932192. DOI: 10.1038/ncomms12142. View

14.

Yang B, Treweek J, Kulkarni R, Deverman B, Chen C, Lubeck E . Single-cell phenotyping within transparent intact tissue through whole-body clearing. Cell. 2014; 158(4):945-958. PMC: 4153367. DOI: 10.1016/j.cell.2014.07.017. View

15.

Conchello J, Lichtman J . Optical sectioning microscopy. Nat Methods. 2005; 2(12):920-31. DOI: 10.1038/nmeth815. View

16.

Zhang C, Liu M, Xing X . Temperature influence on fluorescence intensity and enzyme activity of the fusion protein of GFP and hyperthermophilic xylanase. Appl Microbiol Biotechnol. 2009; 84(3):511-7. DOI: 10.1007/s00253-009-2006-8. View

17.

Hama H, Hioki H, Namiki K, Hoshida T, Kurokawa H, Ishidate F . ScaleS: an optical clearing palette for biological imaging. Nat Neurosci. 2015; 18(10):1518-29. DOI: 10.1038/nn.4107. View

18.

dos Santos A . Thermal effect on Aequorea green fluorescent protein anionic and neutral chromophore forms fluorescence. J Fluoresc. 2011; 22(1):151-4. DOI: 10.1007/s10895-011-0941-0. View

19.

Kim Y, Yang G, Pradhan K, Venkataraju K, Bota M, Garcia Del Molino L . Brain-wide Maps Reveal Stereotyped Cell-Type-Based Cortical Architecture and Subcortical Sexual Dimorphism. Cell. 2017; 171(2):456-469.e22. PMC: 5870827. DOI: 10.1016/j.cell.2017.09.020. View

20.

Watabe-Uchida M, Eshel N, Uchida N . Neural Circuitry of Reward Prediction Error. Annu Rev Neurosci. 2017; 40:373-394. PMC: 6721851. DOI: 10.1146/annurev-neuro-072116-031109. View