Deep-learning microscopy image reconstruction with quality control reveals second-scale rearrangements in RNA polymerase II clusters

BioRxiv : the Preprint Server for Biology
H. HajiabadiLennart Hilbert


Fluorescence microscopy, a central tool of biological research, is subject to inherent trade-offs in experiment design. For instance, image acquisition speed can only be increased in exchange for a lowered signal quality, or for an increased rate of photo-damage to the specimen. Computational denoising can recover some loss of signal, extending the trade-off margin for high-speed imaging. Recently proposed denoising on the basis of neural networks shows exceptional performance but raises concerns of errors typical of neural networks. Here, we present a work-flow that supports an empirically optimized reduction of exposure times, as well as per-image quality control to exclude images with reconstruction errors. We implement this work-flow on the basis of the denoising tool Noise2Void and assess the molecular state and three-dimensional shape of RNA Polymerase II (Pol II) clusters in live zebrafish embryos. Image acquisition speed could be tripled, achieving 2-second time resolution and 350-nanometer lateral image resolution. The obtained data reveal stereotyped events of approximately 10 seconds duration: initially, the molecular mark for initiated Pol II increases, then the mark for active Pol II increases, and finally Pol II c...Continue Reading

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