Simultaneous multi-color optical sectioning fluorescence microscopy with wavelength-coded volume holographic gratings

• Published in: Optics express Vol. 28; no. 25; p. 37177
• Main Authors: Chia, Yu-Hsin, Yeh, J. Andrew, Huang, Yi-You, Luo, Yuan
• Format: Journal Article
• Language: English
• Published: United States 07.12.2020
• ISSN: 1094-4087; 1094-4087
• DOI: 10.1364/OE.409179

Optical sectioning fluorescence microscopy provides high contrast images of volumetric samples and has been widely used for many biological applications. However, simultaneously acquiring multi-color fluorescence images require additional optical elements and devices, which are bulky, wavelength specific, and not cost-effective. In this paper, wavelength-coded volume holographic gratings (WC-VHGs) based optical sectioning fluorescence microscopy is proposed to simultaneously offer multi-color fluorescence images with fine out-of-focus background rejection. Due to wavelength degeneracy, multiplexed WC-VHGs are capable of acquiring multi-wavelength fluorescence images in a single shot, and displaying the laterally separated multi-wavelength images onto CCD. In our system optical sectioning capability is achieved through speckle illumination and HiLo imaging method. To demonstrate imaging characteristics of our system, dual-wavelength fluorescence images of both standard fluorescent microspheres and ex vivo mT/mG mice cardiac tissue are presented. Current results may find important applications in hyperspectral imaging for biomedical research.