1.Institut Fresnel, CNRS, Aix Marseille Univ, Centrale Med, Marseille, France
2.Université de Limoges, CNRS, XLIM, UMR 7252, F-87000 Limoges, France
3.Neurotechnology Center, Department of Biological Sciences, Columbia University, New York, NY 10027, USA
Guillaume Baffou (guillaume.baffou@fresnel.fr)
Published:31 December 2024,
Published Online:11 October 2024,
Received:01 December 2023,
Revised:02 August 2024,
Accepted:01 September 2024
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Chaumet, P. C. et al. Quantitative phase microscopies: accuracy comparison. Light: Science & Applications, 13, 2912-2940 (2024).
Chaumet, P. C. et al. Quantitative phase microscopies: accuracy comparison. Light: Science & Applications, 13, 2912-2940 (2024). DOI: 10.1038/s41377-024-01619-7.
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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