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

2021; American Institute of Physics; Volume: 155; Issue: 16 Linguagem: Inglês

10.1063/5.0063634

1520-9032

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

Biosensors and Analytical Detection

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.