An adaptive microscope for the imaging of biological surfaces
作者机构：Aix Marseille UnivCNRSCentrale MarseilleInstitut FresnelTuring Center for Living SystemsMarseilleFrance
出 版 物：《Light(Science & Applications)》 (光（科学与应用)（英文版）)
年 卷 期：2021年第10卷第11期
页 面：2195-2206页
核心收录：
学科分类：0710[理学-生物学] 07[理学] 070302[理学-分析化学] 0804[工学-仪器科学与技术] 0703[理学-化学] 0702[理学-物理学]
基 金：This work was funded by the following agencies:Agence Nationale de la Recherche(ANR-18-CE13-028,ANR-17-CE30-0007) Excellence Initiative of Aix-Marseille University-A*Midex(capostromex),a French Investissements d’Avenir programme Centre National de la Recherche Scientifique,Mission pour l’Interdisciplinarite Institut Carnot star SATT-SudEst The project leading to this publication has received funding from the“Investissements d’Avenir”French Government program managed by the French National Research Agency(ANR-16-CONV-0001) from Excellence Initiative of Aix-Marseille University-A*MIDEX.
摘 要：Scanning fluorescence microscopes are now able to image large biological samples at high spatial and temporal resolution.This comes at the expense of an increased light dose which is detrimental to fluorophore stability and cell physiology.To highly reduce the light dose,we designed an adaptive scanning fluorescence microscope with a scanning scheme optimized for the unsupervised imaging of cell sheets,which underly the shape of many embryos and organs.The surface of the tissue is first delineated from the acquisition of a very small subset(~0.1%)of sample space,using a robust estimation strategy.Two alternative scanning strategies are then proposed to image the tissue with an improved photon budget,without loss in resolution.The first strategy consists in scanning only a thin shell around the estimated surface of interest,allowing high reduction of light dose when the tissue is curved.The second strategy applies when structures of interest lie at the cell periphery(e.***.adherens junctions).An iterative approach is then used to propagate scanning along cell contours.We demonstrate the benefit of our approach imaging live epithelia from Drosophila melanogaster.On the examples shown,both approaches yield more than a 20-fold reduction in light dose-and up to more than 80-fold-compared to a full scan of the volume.These smart-scanning strategies can be easily implemented on most scanning fluorescent imaging modality.The dramatic reduction in light exposure of the sample should allow prolonged imaging of the live processes under investigation.
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