4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector
Andrea Bucci,
Giorgio Tortarolo,
Marcus Oliver Held,
Luca Bega,
Eleonora Perego,
Francesco Castagnetti,
Irene Bozzoni,
Eli Slenders and
Giuseppe Vicidomini ()
Additional contact information
Andrea Bucci: Istituto Italiano di Tecnologia
Giorgio Tortarolo: Istituto Italiano di Tecnologia
Marcus Oliver Held: Istituto Italiano di Tecnologia
Luca Bega: Istituto Italiano di Tecnologia
Eleonora Perego: Istituto Italiano di Tecnologia
Francesco Castagnetti: Istituto Italiano di Tecnologia
Irene Bozzoni: Istituto Italiano di Tecnologia
Eli Slenders: Istituto Italiano di Tecnologia
Giuseppe Vicidomini: Istituto Italiano di Tecnologia
Nature Communications, 2024, vol. 15, issue 1, 1-12
Abstract:
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 μm2/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.
Date: 2024
References: View references in EconPapers View complete reference list from CitEc
Citations:
Downloads: (external link)
https://www.nature.com/articles/s41467-024-50512-9 Abstract (text/html)
Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.
Export reference: BibTeX
RIS (EndNote, ProCite, RefMan)
HTML/Text
Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-50512-9
Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/
DOI: 10.1038/s41467-024-50512-9
Access Statistics for this article
Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie
More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().