Abstract
The capability to perform multicolor, wide field-of-view (FOV) fluorescence microscopy imaging is important in screening and pathology applications. We developed a microscopic slide-imaging system that can achieve multicolor, wide FOV, fluorescence imaging based on the Talbot effect. In this system, a light-spot grid generated by the Talbot effect illuminates the sample. By tilting the excitation beam, the Talbot-focused spot scans across the sample. The images are reconstructed by collecting the fluorescence emissions that correspond to each focused spot with a relay optics arrangement. The prototype system achieved an FOV of 12 × 10 mm(2) at an acquisition time as fast as 23 s for one fluorescence channel. The resolution is fundamentally limited by spot size, with a demonstrated full-width at half-maximum spot diameter of 1.2 μm. The prototype was used to nimage green fluorescent beads, double-stained human breast cancer SK-BR-3 cells, Giardia lamblia cysts, and the Cryptosporidium parvum oocysts. This imaging method is scalable and simple for implementation of high-speed wide FOV fluorescence microscopy.
Highlights
Due to the high sensitivity of fluorescent probes, fluorescence microscopy plays a vital role in modern clinical diagnosis and biological research
3.4 Multicolor fluorescence imaging of enteric parasites We further demonstrated the imaging of Giardia lamblia cysts and Cryptosporidium parvum oocytes with our multicolor FTM prototype
We have developed a multicolor FTM prototype based on the Talbot effect that can achieve wide FOV, multicolor fluorescence imaging of samples prepped on glass slides
Summary
Due to the high sensitivity of fluorescent probes, fluorescence microscopy plays a vital role in modern clinical diagnosis and biological research. This modification can reduce the full-field imaging time by reducing the total number of step-stops required To address this speed limitation, there have been a number of published studies that have explored the prospect of using point-scan methods in place of a conventional microscope to accomplish wide FOV fluorescence imaging [6,7,8,9,10]. Using these methods, a grid of focused spots is generated by a grid of microlenses [6, 7, 10], a diffractive optical element [11], or a hologram plate [8].
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