4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector.

Details

Serval ID
serval:BIB_D135DAA5F232
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector.
Journal
Nature communications
Author(s)
Bucci A., Tortarolo G., Held M.O., Bega L., Perego E., Castagnetti F., Bozzoni I., Slenders E., Vicidomini G.
ISSN
2041-1723 (Electronic)
ISSN-L
2041-1723
Publication state
Published
Issued date
23/07/2024
Peer-reviewed
Oui
Volume
15
Number
1
Pages
6188
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
Single-particle tracking techniques enable investigation of the complex functions and interactions of individual particles in biological environments. Many such techniques exist, each demonstrating trade-offs between spatiotemporal resolution, spatial and temporal range, technical complexity, and information content. To mitigate these trade-offs, we enhanced a confocal laser scanning microscope with an asynchronous read-out single-photon avalanche diode array detector. This detector provides an image of the particle's emission, precisely reflecting its position within the excitation volume. This localization is utilized in a real-time feedback system to drive the microscope scanning mechanism and ensure the particle remains centered inside the excitation volume. As each pixel is an independent single-photon detector, single-particle tracking is combined with fluorescence lifetime measurement. Our system achieves 40 nm lateral and 60 nm axial localization precision with 100 photons and sub-millisecond temporal sampling for real-time tracking. Offline tracking can refine this precision to the microsecond scale. We validated the system's spatiotemporal resolution by tracking fluorescent beads with diffusion coefficients up to 10 μm <sup>2</sup> /s. Additionally, we investigated the movement of lysosomes in living SK-N-BE cells and measured the fluorescence lifetime of the marker expressed on a membrane protein. We expect that this implementation will open other correlative imaging and tracking studies.
Pubmed
Web of science
Open Access
Yes
Create date
26/07/2024 12:56
Last modification date
13/08/2024 6:48
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