The side lobes of Bessel beam will create significant out-of-focus background when scanned in light-sheet fluorescence microscopy (LSFM), limiting the axial resolution of the imaging system. Baoli Yao’s research team recently proposed to overcome this issue by scanning the sample twice with zeroth-order Bessel beam and another type of propagation-invariant beam, complementary to the zeroth-order Bessel beam, which greatly reduces the out-of-focus background created in the first scan. The axial resolution can be improved from 1.68 mu m of the Bessel light-sheet to 1.07 mu m by subtraction of the two scanned images across a whole field-of-view of up to 300 mu m x 200 mu m x 200 mu m. The optimization procedure to create the complementary beam is described in detail and it is experimentally generated with a spatial light modulator. The imaging performance is validated experimentally with fluorescent beads as well as eGFP-labeled mouse brain neurons.
（Original research article "Journal of Biophotonics Vol. 12, Issue 1, UNSP e201800094 (2019) https://onlinelibrary.wiley.com/doi/abs/10.1002/jbio.201800094"）
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