Four-dimensional multi-particle tracking in living cells based on lifetime imaging
Research on dynamic events in living cells, such as intracellular transportation, is important for understanding cell functions. As movements occur within cells, the microenvironment of the moving vesicles or biomacromolecules may affect the behavior of them. Herein, we propose a method of simultane...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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De Gruyter
2022-03-01
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| Series: | Nanophotonics |
| Subjects: | |
| Online Access: | https://doi.org/10.1515/nanoph-2021-0681 |
| _version_ | 1797947879291092992 |
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| author | Chen Danni Li Heng Yu Bin Qu Junle |
| author_facet | Chen Danni Li Heng Yu Bin Qu Junle |
| author_sort | Chen Danni |
| collection | DOAJ |
| description | Research on dynamic events in living cells, such as intracellular transportation, is important for understanding cell functions. As movements occur within cells, the microenvironment of the moving vesicles or biomacromolecules may affect the behavior of them. Herein, we propose a method of simultaneously monitoring changes in spatial positions and the local environment related to the fluorescence lifetime, i.e., four-dimensional (4D) multi-particle parallel-tracking in living cells. Based on double-helix point spread function (DH-PSF) microscopy and streak camera, the method combines three-dimensional (3D) localization methods and fluorescence lifetime imaging. By modifying the PSF of the system, the 3D positions and fluorescence lifetime information for several molecules within a depth of a few microns can be acquired simultaneously from a single snapshot. The feasibility of this method is verified by simulating the real-time tracking of a single particle with a given trajectory. In addition, a proof-of-concept 4D tracking system based on the DH-PSF and streak camera was built. The experimental results show that the 3D localization and lifetime precision are σ(x, y, z) = (26 nm, 35 nm, 53 nm) and σ(τ) = 103 ps, respectively, and the effective depth of field is approximately 4 μm. Finally, intracellular endocytosis in a living cell was observed using the system, which demonstrated the successful 4D tracking of two microspheres moving within an axial depth of 4 μm. This work opens a new perspective for research of dynamic processes, by providing information about the chemical (microenvironments) and physical (positions) changes of moving targets in living cells. |
| first_indexed | 2024-04-10T21:34:41Z |
| format | Article |
| id | doaj.art-4165b4cdd9c94b09a311a595a201beed |
| institution | Directory Open Access Journal |
| issn | 2192-8614 |
| language | English |
| last_indexed | 2024-04-10T21:34:41Z |
| publishDate | 2022-03-01 |
| publisher | De Gruyter |
| record_format | Article |
| series | Nanophotonics |
| spelling | doaj.art-4165b4cdd9c94b09a311a595a201beed2023-01-19T12:46:59ZengDe GruyterNanophotonics2192-86142022-03-011181537154710.1515/nanoph-2021-0681Four-dimensional multi-particle tracking in living cells based on lifetime imagingChen Danni0Li Heng1Yu Bin2Qu Junle3Center for Biomedical Optics and Photonics (CBOP) & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Guangdong Province and Ministry of Education, Shenzhen University, Shenzhen, 518060, ChinaCenter for Biomedical Optics and Photonics (CBOP) & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Guangdong Province and Ministry of Education, Shenzhen University, Shenzhen, 518060, ChinaCenter for Biomedical Optics and Photonics (CBOP) & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Guangdong Province and Ministry of Education, Shenzhen University, Shenzhen, 518060, ChinaCenter for Biomedical Optics and Photonics (CBOP) & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Guangdong Province and Ministry of Education, Shenzhen University, Shenzhen, 518060, ChinaResearch on dynamic events in living cells, such as intracellular transportation, is important for understanding cell functions. As movements occur within cells, the microenvironment of the moving vesicles or biomacromolecules may affect the behavior of them. Herein, we propose a method of simultaneously monitoring changes in spatial positions and the local environment related to the fluorescence lifetime, i.e., four-dimensional (4D) multi-particle parallel-tracking in living cells. Based on double-helix point spread function (DH-PSF) microscopy and streak camera, the method combines three-dimensional (3D) localization methods and fluorescence lifetime imaging. By modifying the PSF of the system, the 3D positions and fluorescence lifetime information for several molecules within a depth of a few microns can be acquired simultaneously from a single snapshot. The feasibility of this method is verified by simulating the real-time tracking of a single particle with a given trajectory. In addition, a proof-of-concept 4D tracking system based on the DH-PSF and streak camera was built. The experimental results show that the 3D localization and lifetime precision are σ(x, y, z) = (26 nm, 35 nm, 53 nm) and σ(τ) = 103 ps, respectively, and the effective depth of field is approximately 4 μm. Finally, intracellular endocytosis in a living cell was observed using the system, which demonstrated the successful 4D tracking of two microspheres moving within an axial depth of 4 μm. This work opens a new perspective for research of dynamic processes, by providing information about the chemical (microenvironments) and physical (positions) changes of moving targets in living cells.https://doi.org/10.1515/nanoph-2021-0681fluorescence lifetime microscopymulti-particle trackingsingle particle tracking |
| spellingShingle | Chen Danni Li Heng Yu Bin Qu Junle Four-dimensional multi-particle tracking in living cells based on lifetime imaging Nanophotonics fluorescence lifetime microscopy multi-particle tracking single particle tracking |
| title | Four-dimensional multi-particle tracking in living cells based on lifetime imaging |
| title_full | Four-dimensional multi-particle tracking in living cells based on lifetime imaging |
| title_fullStr | Four-dimensional multi-particle tracking in living cells based on lifetime imaging |
| title_full_unstemmed | Four-dimensional multi-particle tracking in living cells based on lifetime imaging |
| title_short | Four-dimensional multi-particle tracking in living cells based on lifetime imaging |
| title_sort | four dimensional multi particle tracking in living cells based on lifetime imaging |
| topic | fluorescence lifetime microscopy multi-particle tracking single particle tracking |
| url | https://doi.org/10.1515/nanoph-2021-0681 |
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