Dual-modality hyperspectral microscopy for transmission and fluorescence imaging

Abstract

Conventional color imaging provides information in the red, green, and blue (RGB) channels with limited resolution in the spectral domain. This could lead to poor sensitivity in imaging biological samples. In fluorescence imaging, the use of multiple fluorophores is often necessary to label proteins and DNAs for in vivo experiments. Crosstalk between fluorophores can be troublesome because it is challenging to differentiate each fluorophore when their emission spectra are overlapped. To help address these issues, we developed a dual-modality hyperspectral microscopy system that combines hyperspectral imaging and microscopy imaging to provide spatial and spectral information of the samples. The dual-modality feature allows us to study biological samples and fluorescent samples using the same system. We show that applications of the system enable: the identification of different tissue and cell structures; identification of each quantum dot fluorophore, calculation of their relative proportions at each location, and elucidation of their spatial distributions in a mixture sample containing multiple types of quantum dot fluorophores. The results give a brief showcase of how hyperspectral imaging can be useful for biomedical imaging and fluorescence imaging applications.