Label-free superior contrast with c-band ultra-violet extinction microscopy
作者机构：Department of Physics and TechnologyUiT The Arctic University of NorwayTromsøNorway Department of Clinical ScienceIntervention and TechnologyKarolinska InstituteStockholmSweden
出 版 物：《Light(Science & Applications)》 (光（科学与应用)（英文版）)
年 卷 期：2023年第12卷第3期
页 面：435-445页
核心收录：
学科分类：0710[理学-生物学] 070207[理学-光学] 07[理学] 08[工学] 0803[工学-光学工程] 0702[理学-物理学]
基 金：European Union’s Horizon 2020 Marie Skłodowska-Curie Action no.836355 project MitoQuant.Open access funding provided by UiT The Arctic University of Norway(incl University Hospital of North Norway)
主 题：wave. extinction violet
摘 要：In 1934,Frits Zernike demonstrated that it is possible to exploit the sample’s refractive index to obtain superior contrast images of biological *** refractive index contrast of a cell surrounded by media yields a change in the phase and intensity of the transmitted light *** change can be due to either scattering or absorption caused by the *** cells are transparent at visible wavelengths,which means the imaginary component of their complex refractive index,also known as extinction coefficient k,is close to ***,we explore the use of c-band ultra-violet(UVC)light for high-contrast high-resolution label-free microscopy,as k is naturally substantially higher in the UVC than at visible *** differential phase contrast illumination and associated processing,we achieve a 7-to 300-fold improvement in contrast compared to visible-wavelength and UVA differential interference contrast microscopy or holotomography,and quantify the extinction coefficient distribution within liver sinusoidal endothelial *** a resolution down to 215 nm,we are,for the first time in a far-field label-free method,able to image individual fenestrations within their sieve plates which normally requires electron or fluorescence superresolution *** illumination also matches the excitation peak of intrinsically fluorescent proteins and amino acids and thus allows us to utilize autofluorescence as an independent imaging modality on the same setup.
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