• Photonics Research
  • Vol. 10, Issue 5, 1255 (2022)
Jiazhen Zhai1、†, Ruheng Shi1、†, and Lingjie Kong1、2、*
Author Affiliations
  • 1State Key Laboratory of Precision Measurement Technology and Instruments, Department of Precision Instrument, Tsinghua University, Beijing 100084, China
  • 2IDG/McGovern Institute for Brain Research, Tsinghua University, Beijing 100084, China
  • show less
    DOI: 10.1364/PRJ.451895 Cite this Article Set citation alerts
    Jiazhen Zhai, Ruheng Shi, Lingjie Kong. Improving signal-to-background ratio by orders of magnitude in high-speed volumetric imaging in vivo by robust Fourier light field microscopy[J]. Photonics Research, 2022, 10(5): 1255 Copy Citation Text show less

    Abstract

    Fourier light field microscopy (FLFM) shows great potential in high-speed volumetric imaging of biodynamics. However, due to the inherent disadvantage of wide-field illumination, it suffers from intense background, arising from out of the depth-of-field signal and tissue scattered noise. The background will not only deteriorate the image contrast, making quantitative measurement difficult, but also introduce artifacts, especially in functional imaging of the neuronal network activity in vivo. Here, we propose the robust Fourier light field microscopy (RFLFM), which suppresses the background in FLFM by introducing structured illumination and computational reconstruction based on HiLo. The superior performance of RFLFM is verified by volumetric imaging of biological dynamics in larval zebrafish and mouse in vivo, at a volumetric imaging rate up to 33.3 Hz. The statistical results show that the fluorescence background can be significantly depressed, with the signal-to-background ratio improved by orders of magnitude and the whole image contrast improved by as much as 10.4 times. Moreover, we stress that, in functional imaging of neuronal network activity in turbid brain tissues, our system can avoid artifacts resulting from background fluctuations, while conventional light field microscopy fails. As a simple but powerful tool, we anticipate our technique to be widely adopted in robust, high-contrast, high-speed volumetric imaging.
    FWHMaxial=0.54Ks·NAillumination,

    View in Article

    C(ρ)=σ[I(ρ)]I[ρ],(C1)

    View in Article

    SBR=IsignalIbackgroundIbackground.(C2)

    View in Article

    ρX,Y=cov(X,Y)σXσY,(C3)

    View in Article

    Jiazhen Zhai, Ruheng Shi, Lingjie Kong. Improving signal-to-background ratio by orders of magnitude in high-speed volumetric imaging in vivo by robust Fourier light field microscopy[J]. Photonics Research, 2022, 10(5): 1255
    Download Citation