How the knowledge of the image formation can help to improve the image quality and contrast?

Session

Reproducibility of Data Analysis at Scale

Authors

Ms Ratsimandresy Holinirina Dina Miora (2, 1, 3, 4), Prof. Dr. Erich Rohwer (2), Dr. Gurthwin Bosman (2), Prof. Dr. Rainer Heintzmann (1, 3)

Affiliations

1. Friedrich-Schiller-Universität Jena
2. Stellenbosch University
3. Leibniz Institute of Photonic Technology
4. MRC Laboratory of Molecular Biology

Keywords

PSF, deconvolution, fluorescence microscopy

Abstract text

Modelling a realistic representation of the response of an optical system, defined as the point spread function (PSF), is not a straightforward task^[1], but is essential for an accurate image reconstruction^[2]. Here we present four Fourier-based PSF models of wide-field imaging^[3]. The models fit the experimental PSFs of high NA system at a high accuracy. They are also experimentally validated in the deconvolution of C. elegans embryo^[4] and spherical beads^[4] of 2.5 µm in diameter and result with a contrast improvement up to 162 times in the images. The models enable a faster deconvolution convergence.

 

References

[1] Richards, B., & Wolf, E. (1959). Electromagnetic diffraction in optical systems, II. Structure of the image field in an aplanatic system. Proceedings of the Royal Society of London. Series A. Mathematical and Physical Sciences, 253(1274), 358-379.

 [2] Sarder, P., & Nehorai, A. (2006). Deconvolution methods for 3-D fluorescence microscopy images. IEEE signal processing magazine, 23(3), 32-45.

[3] Miora, R. H. D., Rohwer, E., Kielhorn, M., Sheppard, C. J., Bosman, G., & Heintzmann, R. (2023). Calculating Point Spread Functions: Methods, Pitfalls and Solutions. arXiv preprint arXiv:2301.13515.

[4] Griffa, A., Garin, N., & Sage, D. (2010). Comparison of deconvolution software in 3D microscopy: A user point of view—Part 1. GIT Imaging & Microscopy, 12(ARTICLE), 43-45.