Imaging quality enhancement for high numerical aperture confocal scanning fluorescence microscopy by using two beams

The confocal scanning fluorescence microscopy is a powerful and effective tool which makes a great contribution to explore life sciences. Nevertheless, according to light-diffraction theory, the spatial resolution of this device is predicted to near 200 nm. In order to gain an insight into life science, this spatial resolution of the confocal scanning fluorescence microscopy must be designed to break the limit of theory of light diffraction. In this paper, the improvement of the higher spatial resolution for the confocal scanning fluorescence microscopy is introduced by using two illumination beams for activating the sample. The resolution-improving ability of the proposed method has been demonstrated by the comparison of the imaging results between the confocal scanning fluorescence microscopy with the Gaussian illumination beam and the proposed method. © 2023 The Authors