Experimental validation of numerical point spread function calculation including aberration estimation

Author:

Holinirina Dina Miora Ratsimandresy1ORCID,Senftleben Maximilian2ORCID,Abrahamsson Sara3,Rohwer ErichORCID,Heintzmann Rainer14ORCID,Bosman Gurthwin5

Affiliation:

1. Institute of Physical Chemistry and Abbe Center of Photonics Friedrich-Schiller-University Jena

2. KTH Royal Institute of Technology

3. University of California Santa Cruz

4. Leibniz Institute of Photonic Technology

5. National Institute for Theoretical and Computational Sciences (NITheCS)

Abstract

Image reconstruction in fluorescence microscopy is highly sensitive to the accuracy of the impulse response, defined as the point spread function (PSF), of the optical system under which the image to reconstruct was acquired. In our previous work, we developed a MATLAB toolbox for accurately calculating realistic vector Fourier-based PSF accounting for any type of aberrations [arXiv, arXiv:2301.13515 (2023)10.48550/arXiv.2301.13515]. In this work, we present a fundamental experimental validation of these numerical methods. The simulated results are found to fit experimental data under different image acquisition conditions at an accuracy higher than 0.97 in normalized cross-correlation. These methods enable a relative contrast of up to 95%.

Funder

Deutsche Forschungsgemeinschaft

Council for Scientific and Industrial Research, South Africa

Publisher

Optica Publishing Group

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