Ultra-low-cost and high-fidelity NIR-II confocal laser scanning microscope with Bessel beam excitation and SiPM detection.

Abstract

Confocal laser scanning microscopy (CLSM) is one of the most important imaging tools in the biomedical field, and near-infrared-II (NIR-II, 900-1700nm) fluorescence imaging technology has also made fruitful research progress in deep imaging in recent years. The NIR-II based CLSM has problems such as an expensive detector and reduced image resolution caused by long wavelength excitation. Here, by simultaneously using a low-cost silicon photomultiplier (SiPM) as a detector and a Bessel beam as an excitation, we developed an ultra-low-cost and high-fidelity NIR-II confocal laser scanning microscope. The use of SiPM reduces the cost of the NIR-II fluorescence detection module in CLSM, while enabling the detection of ultra-broadband fluorescence signals spanning visible to NIR-II regions. The introduction of the Bessel beam compensates to some extent for the weakening of spatial resolution caused by the increase in the wavelength of light in the NIR region. Experimental results show that the use of the Bessel beam can improve the resolution by 12% when observing thin samples. With the increase of sample thickness, the imaging resolution of the Bessel beam at NIR-II wavelengths is better than that of the Gaussian beam at NIR-I wavelengths at the penetrable depth of the NIR-I light. At deeper depths, the imaging resolution and imaging depth of Bessel beam CLSM is superior to Gaussian beam CLSM at the same excitation power.