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Out-of-focus and scattered light often obscure dim fluorescent fibers in widefield neuronal imaging. We solve this obstacle by spatially modulating the excitation light to illuminate different neuronal structures with different light intensities. In this way, we minimize background and enhance the visibility of neuron fibers in the final image. This method significantly improves fiber contrast while maintaining a fast imaging speed and low cost. Using this widefield targeted illumination setup, we demonstrate confocal quality imaging of complex neurons in the nematode C. elegans.
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Yao Wang, Jia Fan, Samuel Chung, "Targeted illumination on neuronal fibers in widefield microscopy achieves confocal quality neuronal images," Proc. SPIE PC11946, Neural Imaging and Sensing 2022, PC119460M (28 April 2022); https://doi.org/10.1117/12.2626229