• Advanced Photonics Nexus
  • Vol. 2, Issue 1, 016001 (2023)
Liang Qiao1、†, Hongjin Li1、2, Suyi Zhong3, Xinzhu Xu3, Fei Su1、4, Xi Peng1、3, Dayong Jin1、4、*, and Karl Zhanghao1、*
Author Affiliations
  • 1Southern University of Science and Technology, College of Engineering, UTS-SUSTech Joint Research Centre for Biomedical Materials and Devices, Department of Biomedical Engineering, Shenzhen, China
  • 2City University of Hong Kong, Department of Biomedical Engineering, Hong Kong, China
  • 3Peking University, College of Future Technology, Department of Biomedical Engineering, Beijing, China
  • 4University of Technology Sydney, Institute for Biomedical Materials and Devices (IBMD), Faculty of Science, Sydney, Australia
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    DOI: 10.1117/1.APN.2.1.016001 Cite this Article Set citation alerts
    Liang Qiao, Hongjin Li, Suyi Zhong, Xinzhu Xu, Fei Su, Xi Peng, Dayong Jin, Karl Zhanghao. Laterally swept light-sheet microscopy enhanced by pixel reassignment for photon-efficient volumetric imaging[J]. Advanced Photonics Nexus, 2023, 2(1): 016001 Copy Citation Text show less

    Abstract

    In light-sheet fluorescence microscopy, the axial resolution and field of view are mutually constrained. Axially swept light-sheet microscopy (ASLM) can decouple the trade-off, but the confocal detection scheme using a rolling shutter also rejects fluorescence signals from the specimen in the field of interest, which sacrifices the photon efficiency. Here, we report a laterally swept light-sheet microscopy (LSLM) scheme in which the focused beam is first scanned along the axial direction and subsequently laterally swept with the rolling shutter. We show that LSLM can obtain a higher photon efficiency when similar axial resolution and field of view can be achieved. Moreover, based on the principle of image scanning microscopy, applying the pixel reassignment to the LSLM images, hereby named iLSLM, improves the optical sectioning. Both simulation and experimental results demonstrate the higher photon efficiency with similar axial resolution and optical sectioning. Our proposed scheme is suitable for volumetric imaging of specimens that are susceptible to photobleaching or phototoxicity.
    I=|E(x,z;y)|2,

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    E(x,z;y)=F[E(kx.kz)exp(ikyΔy)],

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    E(kx,kz)=exp[(kx2+kz2)/(k0N.A.)2],

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    σISM=(1/σexc2+1/σdet2)1/2,

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    IASLM1(x,yys;zzs)=[o(x,y;zzs)xsIex(xxs,z;yys)]*hdet(x,y;z),

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    ILSLM1(xxs,y;zzs)=[o(x,y;zzs)ysIex(xxs,z;yys)]*hdet(x,y;z),

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    IASLM2(x,yys;z0zs)=IASLM1(x,yys;z0zs)S(x,yys),

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    ILSLM2(xxs,y;z0zs)=ILSLM1(xxs,y;z0zs)S(xxs,y),

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    IASLM(x,y;zs)=ysIASLM2(x,yys;z0zs),

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    ILSLM(x,y;zs)=xsILSLM2(xxs,y;z0zs),

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    IiLSLM(x,y;zs)=xsILSLM2(xxsm,y;z0zs).

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    ϕaxial(x,z)=kΔy(xm2+zn2)/2fscan2,

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    ϕlateral(x,z)=k(xmsinθx+znsinθz),

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    Δx=fscantanθx,Δz=fscantanθz.

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    Liang Qiao, Hongjin Li, Suyi Zhong, Xinzhu Xu, Fei Su, Xi Peng, Dayong Jin, Karl Zhanghao. Laterally swept light-sheet microscopy enhanced by pixel reassignment for photon-efficient volumetric imaging[J]. Advanced Photonics Nexus, 2023, 2(1): 016001
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