Quantitative phase microscopies: accuracy comparison - Archive ouverte HAL
Article Dans Une Revue Light: Science and Applications Année : 2024

Quantitative phase microscopies: accuracy comparison

Patrick C Chaumet
Pierre Bon
Guillaume Maire
  • Fonction : Auteur
Anne Sentenac
  • Fonction : Auteur
Guillaume Baffou

Résumé

Quantitative phase microscopies (QPMs) play a pivotal role in bio-imaging, offering unique insights that complement fluorescence imaging. They provide essential data on mass distribution and transport, inaccessible to fluorescence techniques. Additionally, QPMs are label-free, eliminating concerns of photobleaching and phototoxicity. However, navigating through the array of available QPM techniques can be complex, making it challenging to select the most suitable one for a particular application. This tutorial review presents a thorough comparison of the main QPM techniques, focusing on their accuracy in terms of measurement precision and trueness. We focus on 8 techniques, namely digital holographic microscopy (DHM), cross-grating wavefront microscopy (CGM), which is based on QLSI (quadriwave lateral shearing interferometry), diffraction phase microscopy (DPM), differential phase-contrast (DPC) microscopy, phase-shifting interferometry (PSI) imaging, Fourier phase microscopy (FPM), spatial light interference microscopy (SLIM), and transport-of-intensity equation (TIE) imaging. For this purpose, we used a home-made numerical toolbox based on discrete dipole approximation (IF-DDA). This toolbox is designed to compute the electromagnetic field at the sample plane of a microscope, irrespective of the object's complexity or the illumination conditions. We upgraded this toolbox to enable it to model any type of QPM, and to take into account shot noise. In a nutshell, the results show that DHM and PSI are inherently free from artefacts and rather suffer from coherent noise; In CGM, DPC, DPM and TIE, there is a trade-off between precision and trueness, which can be balanced by varying one experimental parameter; FPM and SLIM suffer from inherent artefacts that cannot be discarded experimentally in most cases, making the techniques not quantitative especially for large objects covering a large part of the field of view, such as eukaryotic cells.

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hal-04757496 , version 1 (29-10-2024)

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Patrick C Chaumet, Pierre Bon, Guillaume Maire, Anne Sentenac, Guillaume Baffou. Quantitative phase microscopies: accuracy comparison. Light: Science and Applications, 2024, 13, pp.288. ⟨10.1038/s41377-024-01619-7⟩. ⟨hal-04757496⟩
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