Design and research of fully automatic push-broom hyperspectral microscopic imaging system

To apply hyperspectral technology to the field of microscopic imaging more conveniently, we designed and built a fully automatic push-broom hyperspectral microscopic imaging system. In this system, an inverted microscope was designed as the main body, a prism-grating component was used for spectrum splitting, a high precision two-dimensional motorized stage was applied for a push-broom. A motor focus module was used to control the focus, and a hyperspectral microscopic image was collected through a highly sensitive sCMOS scientific camera. The system has the advantages of low cost, easy installation and adjustment, real-time focusing and large-field-of-view imaging. The spectral range of the system is from 420 nm to 800 nm to meet the spectrum detection requirements of most biological samples. The spectral resolution was better than 3.5 nm, and the spatial resolution was better than 0.87 μm through the monochromatic collimated light scanning calibration method. Then, the HE-stained breast cancer pathological slices was as the research object. The samples were investigated and compared using passive and active focusing for push-broom imaging. The advantages and disadvantages of the two focusing methods were analyzed and summarized. The results showed that both methods can meet the needs of large-field-of-view imaging, but active focus imaging is faster and clearer, and is more suitable for push-broom hyperspectral microscopy imaging systems. Through the design and research of a fully automatic push-broom hyperspectral microscopy imaging system, real-time focusing in hyperspectral microscopic imaging was realized and 3.25 mm×3.25 mm field of view imaging of biological samples with a 40X objective lens was achieved. This system could be beneficial for promoting the application of hyperspectral technology in the biomedical field.