1 南昌大学信息工程学院, 江西 南昌 330031
2 中国科学院上海应用物理研究所物理生物实验室, 上海 201800
荧光辐射差分(FED)显微术利用实心光斑扫描的一幅共聚焦图像减去空心光斑扫描的负共聚焦图像,实现了超分辨成像。使用简单的环形光瞳滤波器提高了无变形FED的成像分辨率。在空心焦斑光路上使用合适的环形光瞳滤波器,压缩空心焦斑的尺寸;同时,在实心焦斑的光路上允许使用更高数值孔径的孔径光阑,获得更小的、与空心焦斑相匹配的实心光斑分布:两方面作用下,提高了FED的空间分辨率。
显微 空间分辨率 环形光瞳 荧光辐射差分显微技术 超分辨成像
Author Affiliations
Abstract
1 School of Information Engineering, Nanchang University, Nanchang 330031, China
2 Division of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China
3 Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, Shanghai 201800, China
The fluorescence from the out-of-focus region excited by the sidelobes of a Bessel beam is the major concern for 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 sidelobes and enhanced sectioning ability with improved contrast are demonstrated.
180.2520 Fluorescence microscopy 170.6900 Three-dimensional microscopy 100.2980 Image enhancement Chinese Optics Letters
2018, 16(11): 111801
