Fiber-bundle illumination: realizing high-degree time-multiplexed multifocal multiphoton microscopy with simplicity
Abstract
High-degree time-multiplexed multifocal multiphoton microscopy was expected to provide a facile path to scanningless optical-sectioning and the fast imaging of dynamic three-dimensional biological systems. However, physical constraints on typical time multiplexing devices, arising from diffraction in the free-space propagation of light waves, lead to significant manufacturing difficulties and have prevented the experimental realization of high-degree time multiplexing. To resolve this issue, we have developed a novel method using optical fiber bundles of various lengths to confine the diffraction of propagating light waves and to create a time multiplexing effect. Through this method, we experimentally demonstrate the highest degree of time multiplexing ever achieved in multifocal multiphoton microscopy (~50 times larger than conventional approaches), and hence the potential of using simply-manufactured devices for scanningless optical sectioning of biological systems.
Additional Information
© 2018 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 04 March 2014; Accepted 24 September 2018; Published 05 October 2018. The authors would like to acknowledge Mr. Yun Mou of California Institute of Technology for his kind instructions in selection and usage of fluorescent dyes. C.G. acknowledges the support from Ellison Medical Foundation, funds from Ministry of Science and Technology, Taiwan (104-2112-M-001-043-MY3, 106-2627-M-001-005, 106-2918-I-001-005, 107-2112-M-001-040-MY3, and 107-2119-M-001-039), and fund from Academia Sinica, Taiwan (AS-105-TP-A04). Y.H. acknowledges the support from the Basic Science Research Program through the National Research Foundation of Korea, funded by the Ministry of Education, Science and Technology (NRF-2015R1D1A1A09060875). Support from the National Science Foundation (CHE-1214123) to GAB is gratefully acknowledged. Author Contributions: M.A., G.B., C.G., D.H., Y.H. and J.Y. wrote the main manuscript text and Supplementary Information. D.H., S.K., Y.S., C.Y. and J.Y. prepared Figure 1. S.K., Y.S. and J.Y. prepared Figures 2 and 3, Supplementary Figures 1–3, and Supplementary Movie 2. C.Y. and J.Y. prepared Supplementary Movie 1. The authors declare no competing interests.Attached Files
Published - 41598_2018_Article_33286.pdf
Supplemental Material - 41598_2018_33286_MOESM1_ESM.avi
Supplemental Material - 41598_2018_33286_MOESM2_ESM.mov
Supplemental Material - 41598_2018_33286_MOESM3_ESM.pdf
Supplemental Material - 41598_2018_33286_MOESM4_ESM.tex
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Additional details
- PMCID
- PMC6173774
- Eprint ID
- 90243
- Resolver ID
- CaltechAUTHORS:20181011-115949691
- Ellison Medical Foundation
- 104-2112-M-001-043-MY3
- Ministry of Science and Technology (Taipei)
- 106-2627-M-001-005
- Ministry of Science and Technology (Taipei)
- 106-2918-I-001-005
- Ministry of Science and Technology (Taipei)
- 107-2112-M-001-040-MY3
- Ministry of Science and Technology (Taipei)
- 107-2119-M-001-039
- Ministry of Science and Technology (Taipei)
- AS-105-TP-A04
- Academia Sinica
- NRF-2015R1D1A1A09060875
- National Research Foundation of Korea
- CHE-1214123
- NSF
- Created
-
2018-10-11Created from EPrint's datestamp field
- Updated
-
2021-11-16Created from EPrint's last_modified field
- Caltech groups
- Astronomy Department, Division of Geological and Planetary Sciences