Long-working-distance High-collection-efficiency Three-photon Microscopy for Long-term Imaging of Zebrafish and Organoids

Overview

Journal iScience

Publisher Cell Press

Date 2024 Aug 26

PMID 39184441

Authors

Peng Deng

Shoupei Liu

Yaoguang Zhao

Xinxin Zhang

Yufei Kong

Linlin Liu

Yujie Xiao

Shasha Yang

Jiahao Hu

Jixiong Su

Ang Xuan

Jinhong Xu

Huijuan Li

Xiaoman Su

Jingchuan Wu

Yuli Jiang

Yu Mu

Zhicheng Shao

Cihang Kong

Bo Li

Affiliations

Soon will be listed here.

Abstract

Zebrafish and organoids, crucial for complex biological studies, necessitate an imaging system with deep tissue penetration, sample protection from environmental interference, and ample operational space. Traditional three-photon microscopy is constrained by short-working-distance objectives and falls short. Our long-working-distance high-collection-efficiency three-photon microscopy (LH-3PM) addresses these challenges, achieving a 58% fluorescence collection efficiency at a 20 mm working distance. LH-3PM significantly outperforms existing three-photon systems equipped with the same long working distance objective, enhancing fluorescence collection and dramatically reducing phototoxicity and photobleaching. These improvements facilitate accurate capture of neuronal activity and an enhanced detection of activity spikes, which are vital for comprehensive, long-term imaging. LH-3PM's imaging of epileptic zebrafish not only showed sustained neuron activity over an hour but also highlighted increased neural synchronization and spike numbers, marking a notable shift in neural coding mechanisms. This breakthrough paves the way for new explorations of biological phenomena in small model organisms.

References

Makale M, McElroy M, OBrien P, Hoffman R, Guo S, Bouvet M . Extended-working-distance multiphoton micromanipulation microscope for deep-penetration imaging in live mice and tissue. J Biomed Opt. 2009; 14(2):024032. PMC: 3098575. DOI: 10.1117/1.3103783. View

Chow D, Sinefeld D, Kolkman K, Ouzounov D, Akbari N, Tatarsky R . Deep three-photon imaging of the brain in intact adult zebrafish. Nat Methods. 2020; 17(6):605-608. PMC: 7359951. DOI: 10.1038/s41592-020-0819-7. View

Redman W, Wolcott N, Montelisciani L, Luna G, Marks T, Sit K . Long-term transverse imaging of the hippocampus with glass microperiscopes. Elife. 2022; 11. PMC: 9249394. DOI: 10.7554/eLife.75391. View

Barson D, Hamodi A, Shen X, Lur G, Constable R, Cardin J . Simultaneous mesoscopic and two-photon imaging of neuronal activity in cortical circuits. Nat Methods. 2019; 17(1):107-113. PMC: 6946863. DOI: 10.1038/s41592-019-0625-2. View

Yu C, Yu Y, Adsit L, Chang J, Barchini J, Moberly A . The Cousa objective: a long-working distance air objective for multiphoton imaging in vivo. Nat Methods. 2023; 21(1):132-141. PMC: 10776402. DOI: 10.1038/s41592-023-02098-1. View

Combs C, Smirnov A, Riley J, Gandjbakhche A, Knutson J, Balaban R . Optimization of multiphoton excitation microscopy by total emission detection using a parabolic light reflector. J Microsc. 2007; 228(Pt 3):330-7. DOI: 10.1111/j.1365-2818.2007.01851.x. View

Tallapragada N, Cambra H, Wald T, Keough Jalbert S, Abraham D, Klein O . Inflation-collapse dynamics drive patterning and morphogenesis in intestinal organoids. Cell Stem Cell. 2021; 28(9):1516-1532.e14. PMC: 8419000. DOI: 10.1016/j.stem.2021.04.002. View

Laissue P, Alghamdi R, Tomancak P, Reynaud E, Shroff H . Assessing phototoxicity in live fluorescence imaging. Nat Methods. 2017; 14(7):657-661. DOI: 10.1038/nmeth.4344. View

Devinsky O, Vezzani A, OBrien T, Jette N, Scheffer I, de Curtis M . Epilepsy. Nat Rev Dis Primers. 2018; 4:18024. DOI: 10.1038/nrdp.2018.24. View

10.

Corsini N, Knoblich J . Human organoids: New strategies and methods for analyzing human development and disease. Cell. 2022; 185(15):2756-2769. DOI: 10.1016/j.cell.2022.06.051. View

11.

Shang C, Wang Y, Zhao M, Fan Q, Zhao S, Qian Y . Real-time analysis of large-scale neuronal imaging enables closed-loop investigation of neural dynamics. Nat Neurosci. 2024; 27(5):1014-1018. DOI: 10.1038/s41593-024-01595-6. View

12.

Schneider C, Rasband W, Eliceiri K . NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012; 9(7):671-5. PMC: 5554542. DOI: 10.1038/nmeth.2089. View

13.

Burrows D, Samarut E, Liu J, Baraban S, Richardson M, Meyer M . Imaging epilepsy in larval zebrafish. Eur J Paediatr Neurol. 2020; 24:70-80. PMC: 7035958. DOI: 10.1016/j.ejpn.2020.01.006. View

14.

Yamamoto S, Kanca O, Wangler M, Bellen H . Integrating non-mammalian model organisms in the diagnosis of rare genetic diseases in humans. Nat Rev Genet. 2023; 25(1):46-60. DOI: 10.1038/s41576-023-00633-6. View

15.

Schneider S, Lee J, Mathis M . Learnable latent embeddings for joint behavioural and neural analysis. Nature. 2023; 617(7960):360-368. PMC: 10172131. DOI: 10.1038/s41586-023-06031-6. View

16.

Xiao Y, Deng P, Zhao Y, Yang S, Li B . Three-photon excited fluorescence imaging in neuroscience: From principles to applications. Front Neurosci. 2023; 17:1085682. PMC: 9986337. DOI: 10.3389/fnins.2023.1085682. View

17.

Terada S, Kobayashi K, Ohkura M, Nakai J, Matsuzaki M . Super-wide-field two-photon imaging with a micro-optical device moving in post-objective space. Nat Commun. 2018; 9(1):3550. PMC: 6120955. DOI: 10.1038/s41467-018-06058-8. View

18.

Combs C, Smirnov A, Chess D, McGavern D, Schroeder J, Riley J . Optimizing multiphoton fluorescence microscopy light collection from living tissue by noncontact total emission detection (epiTED). J Microsc. 2010; 241(2):153-61. PMC: 3518454. DOI: 10.1111/j.1365-2818.2010.03411.x. View

19.

Turrini L, Roschi L, De Vito G, Pavone F, Vanzi F . Imaging Approaches to Investigate Pathophysiological Mechanisms of Brain Disease in Zebrafish. Int J Mol Sci. 2023; 24(12). PMC: 10298537. DOI: 10.3390/ijms24129833. View

20.

Hasani H, Sun J, Zhu S, Rong Q, Willomitzer F, Amor R . Whole-brain imaging of freely-moving zebrafish. Front Neurosci. 2023; 17:1127574. PMC: 10150962. DOI: 10.3389/fnins.2023.1127574. View