Label-free imaging of red blood cells and oxygenation with color third-order sum-frequency generation microscopy  

作　　者：Julia Ferrer Ortas Pierre Mahou Sophie Escoto Chiara Stringari Nicolas BDavid Laure Bally-Cuif Nicolas Dray Michel Negrerie Willy Supatto Emmanuel Beaurepaire 

机构地区：[1]Laboratory for Optics and Biosciences,CNRS,INSERM,Ecole polytechnique,IP Paris,91128 Palaiseau,France [2]Zebrafish Neurogenetics Unit,team supported by Ligue Nationale contre le Cancer,Institut Pasteur,CNRS,75015 Paris,France

出　　处：《Light(Science & Applications)》2023年第2期168-182,共15页光（科学与应用)（英文版）

摘　　要：Mapping red blood cells(RBCs)flow and oxygenation is of key importance for analyzing brain and tissue physiology.Current microscopy methods are limited either in sensitivity or in spatio-temporal resolution.In this work,we introduce a novel approach based on label-free third-order sum-frequency generation(TSFG)and third-harmonic generation(THG)contrasts.First,we propose a novel experimental scheme for color TSFG microscopy,which provides simultaneous measurements at several wavelengths encompassing the Soret absorption band of hemoglobin.We show that there is a strong three-photon(3P)resonance related to the Soret band of hemoglobin in THG and TSFG signals from zebrafish and human RBCs,and that this resonance is sensitive to RBC oxygenation state.We demonstrate that our color TSFG implementation enables specific detection of flowing RBCs in zebrafish embryos and is sensitive to RBC oxygenation dynamics with single-cell resolution and microsecond pixel times.Moreover,it can be implemented on a 3P microscope and provides label-free RBC-specific contrast at depths exceeding 600μm in live adult zebrafish brain.Our results establish a new multiphoton contrast extending the palette of deep-tissue microscopy.

关 键 词：ZEBRA EXCEEDING free 

分 类 号：TH742[机械工程—光学工程]