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Particles 3D tracking with large axial depth by using the 2π-DH-PSF

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Abstract

We propose a ${{2}}\pi$-double-helix point spread function (${{2}}\pi$-DH-PSF) using the Fresnel zone approach that can rotate ${{2}}\pi$ rad. When 16 Fresnel zones are used, the particles can be tracked in the axial range of 10 µm in a ${{100}} \times$ microscopy imaging system (${\rm{NA}} = {1.4}$, $\lambda = {{514}}\;{\rm{nm}}$). We measured the diffusion coefficient of nanospheres in different concentrations of glycerol with the ${{2}}\pi$-DH-PSF, and the error between the measured results and theoretical value was within 10%, indicating the superior performance of ${{2}}\pi$-DH-PSF in 3D localization imaging of nanoparticles. When combined with the defocus phase, the rotation angle can reach ${{4}}\pi$ rad, four times that of the conventional DH-PSF.

© 2021 Optical Society of America

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Supplementary Material (6)

NameDescription
Visualization 1       The sample was moved 0.25µm/step along the Z-axis from -10µm to 10.25µm by using a Nano displacement platform (OPLAN Nano Z100).
Visualization 2       The sample was moved 0.5µm/step along the Z-axis from -15.5µm to 14.5µm by using a Nano displacement platform (OPLAN Nano Z100).
Visualization 3       The movement process of fluorescent microspheres on the cell membrane of Hela cells.
Visualization 4       The movement process of fluorescent microspheres inside Hela cells.
Visualization 5       The movement process of fluorescent microspheres inside floating Hela cells.
Visualization 6       The three-dimensional trajectory of fluorescent microspheres in saliva.

Data Availability

All data underlying the results presented in this Letter are available herein.

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Equations (10)

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