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.
Patrick C.Chaumet、Pierre Bon、Guillaume Maire、Anne Sentenac、Guillaume Baffou
国家科技期刊平台
NSTL
万方数据
