The fluorescence from out-of-focus region excited by the side lobes of Bessel beam is the major concerns for the light-sheet fluorescence microscopy (LSFM) with Bessel beam plane illumination. Here, we propose a method of applying the subtractive imaging to overcome the limitation of the conventional LSFM with Bessel beam plane illumination. In the proposed method, the sample is imaged twice by line-scanning using the extended solid Bessel beam and the ring-like Bessel beam. By subtracting between the two images with similar out-of focus blur, the improved image quality with the suppression of the Bessel beam side lobes and enhanced sectioning ability with improved contrast is demonstrated.
Deng Suhui, Xiao Yiping, Hu Jie, Chen Jianfang, Wang Yuhao, Liu Mingping. Side lobe supression in light-sheet fluorescence microscopy with Bessel beam plane illumination using subtractive imaging[J]. Chinese Optics Letters, 2018, 16(11): .
