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Deformable mirror based optimal PSF engineering for 3D super-resolution imaging

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Abstract

Point spread function (PSF) engineering is an important technique to encode the properties (e.g., 3D positions, color, and orientation) of a single molecule in the shape of the PSF, often with the help of a programmable phase modulator. A deformable mirror (DM) is currently the most widely used phase modulator for fluorescence detection as it shows negligible photon loss. However, it relies on careful calibration for precise wavefront control. Therefore, design of an optimal PSF not only relies on the theoretical calculation of the maximum information content, but also the physical behavior of the phase modulator, which is often ignored during the optimization process. Here, we develop a framework for PSF engineering which could generate a device specific optimal PSF for 3D super-resolution imaging using a DM. We use our method to generate two types of PSFs with depths of field comparable to the widely used astigmatism and tetrapod PSFs, respectively. We demonstrate the superior performance of the DM specific optimal PSF over the conventional astigmatism and tetrapod PSF both theoretically and experimentally.

© 2022 Optica Publishing Group

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Supplementary Material (5)

NameDescription
Code 1       Example and source code for DM based PSF optimization
Supplement 1       Supplementary Information
Visualization 1       Visualization 1 Animated 3D view of the NPC In Figure 3b
Visualization 2       Visualization 2 Animated 3D view of a representative single NPC in Figure 3b. Colorbar: [-320nm -180nm]. Scale bar: 100nm.
Visualization 3       Visualization 3 Animated 3D view of the whole nucleus In Figure 5

Data availability

The code used in this work is available in Ref. [23].

23. S. Fu, L. Zhou, Y. He, X. Liu, X. Hao, and Y. Li, “Deformable mirror based optimal PSF engineering for 3D super-resolution Imaging,” GitHub (2021) [accessed 6 June 2022], https://github.com/Li-Lab-SUSTech/DM_based_PSF_optimization.

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Figures (5)

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Equations (2)

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