Leveraging lifetime information to perform real-time 3D single-particle tracking in noisy environments

Tian Zhao; Joseph S Beckwith; M Junaid Amin; Marcell Pálmai; Preston T Snee; Ming Tien; Haw Yang

doi:10.1063/5.0063634

Leveraging lifetime information to perform real-time 3D single-particle tracking in noisy environments

J Chem Phys. 2021 Oct 28;155(16):164201. doi: 10.1063/5.0063634.

Authors

Tian Zhao¹, Joseph S Beckwith¹, M Junaid Amin¹, Marcell Pálmai², Preston T Snee², Ming Tien³, Haw Yang¹

Affiliations

¹ Department of Chemistry, Princeton University, Princeton, New Jersey 08544, USA.
² Department of Chemistry, University of Illinois at Chicago, Chicago, Illinois 60607-7061, USA.
³ Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.

PMID: 34717352
DOI: 10.1063/5.0063634

Abstract

A microscopy platform that leverages the arrival time of individual photons to enable 3D single-particle tracking of fast-moving (translational diffusion coefficient of ≃3.3 µm²/s) particles in high-background environments is reported here. It combines a hardware-based time-gating module, which enables the rate of photon processing to be as high as 100 MHz, with a two-photon-excited 3D single-particle tracking confocal microscope to enable high sample penetration depth. Proof-of-principle experiments where single quantum dots are tracked in solutions containing dye-stained cellulose, are shown with tracking performance markedly improved using the hardware-based time-gating module. Such a microscope design is anticipated to be of use to a variety of communities who wish to track single particles in cellular environments, which commonly have high fluorescence and scattering background.