Deep learning enhanced light sheet fluorescence microscopy for in vivo 4D imaging of zebrafish heart beating

Time-resolved volumetric fluorescence imaging over an extended duration with high spatial/temporal resolution is a key driving force in biomedical research for investigating spatial-temporal dynamics at organism-level systems, yet it remains a major challenge due to the trade-off among imaging speed, light exposure, illumination power, and image quality. Here, we present a deep-learning enhanced light sheet fluorescence microscopy (LSFM) approach that addresses the restoration of rapid volumetric time-lapse imaging with less than 0.03% light exposure and 3.3% acquisition time compared to a typical standard acquisition. We demonstrate that the convolutional neural network (CNN)-transformer network developed here, namely U-net integrated transformer (UI-Trans), successfully achieves the mitigation of complex noise-scattering-coupled degradation and outperforms state-of-the-art deep learning networks, due to its capability of faithfully learning fine details while comprehending complex global features. With the fast generation of appropriate training data via flexible switching between confocal line-scanning LSFM (LS-LSFM) and conventional LSFM, this method achieves a three- to five-fold signal-to-noise ratio (SNR) improvement and ~1.8 times contrast improvement in ex vivo zebrafish heart imaging and long-term in vivo 4D (3D morphology + time) imaging of heartbeat dynamics at different developmental stages with ultra-economical acquisitions in terms of light dosage and acquisition time.