Mirror-enhanced scanning light-field microscopy for long-term high-speed 3D imaging with isotropic resolution

Abstract

Various biological behaviors can only be observed in 3D at high speed over the long term with low phototoxicity. Light-field microscopy (LFM) provides an elegant compact solution to record 3D information in a tomographic manner simultaneously, which can facilitate high photon efficiency. However, LFM still suffers from the missing-cone problem, leading to degraded axial resolution and ringing effects after deconvolution. Here, we propose a mirror-enhanced scanning LFM (MiSLFM) to achieve long-term high-speed 3D imaging at super-resolved axial resolution with a single objective, by fully exploiting the extended depth of field of LFM with a tilted mirror placed below samples. To establish the unique capabilities of MiSLFM, we performed extensive experiments, we observed various organelle interactions and intercellular interactions in different types of photosensitive cells under extremely low light conditions. Moreover, we demonstrated that superior axial resolution facilitates more robust blood cell tracking in zebrafish larvae at high speed.

Shedding light on biology and healthcare—preface to the special issue on Biomedical Optics

“…To address the missing cone problem in conventional light-field microscopy (LFM), Xiong et al from Tsinghua University developed mirror-enhanced LFM, which realizes long-term high-speed imaging with isotropic 3D resolution. The technique has been applied to the imaging of various cellular organelle interactions and stable 3D blood cell tracking in zebrafish larvae at volume rates as high as 18 Hz 5 . Zhao et al from Beihang University presented halftone spatial frequency domain imaging (SFDI), realizing kilohertz high-speed (two orders of magnitude faster than the state-of-the-art technique) label-free imaging by using DMD for modulation of illumination.…”

Noise-robust phase-space deconvolution for light-field microscopy

“…The information can be used to recover the 3D volume of the sample through deconvolution algorithms, making LFM popular in various biological applications, especially for large-scale neural activities in 3D observation. 1 – 8 Furthermore, some hardware improvement schemes based on LFM, such as scanning LFM (sLFM), 9 , 10 mirror-enhanced sLFM, 11 confocal LFM, 12 etc., have also been proposed.…”

Background inhibited and speed-loss-free volumetric imaging in vivo based on structured-illumination Fourier light field microscopy

“…On the contrary, the recently developed light field microscopy (LFM) has shown great advantages in fast volumetric imaging ( Levoy et al, 2006 ; Broxton et al, 2013 ; Prevedel et al, 2014 ; Wagner et al, 2021 ; Wu et al, 2021 ). Its super volumetric imaging capability has been used in a variety of applications ( Prevedel et al, 2014 ; Pégard et al, 2016 ; Li et al, 2019 ; Chen et al, 2020 ; Sims et al, 2020 ; Xiong et al, 2021 ; Zhang et al, 2021a , b ), especially in neuroscience research in vivo . In recent years, LFM has developed rapidly.…”