We present a novel single-pixel imaging technique that simultaneously images fluorescence and quantitative phase of an object. To extract simultaneously co-registered fluorescence and phase images, the object is illuminated by a pair of spatially coherent monochromatic laser beams with a difference in illumination spatial frequency that is swept linearly in time. One of the beams is stationary – serving as a reference beam – and propagated along the optic axis. The other beam scans through the full range of transverse spatial frequencies supported by the illumination optic – sweeping the crossing angle of the two beams incident on the specimen as a function of time. The scanned beam also has a temporal modulation carrier frequency that allows the extraction of the products of interfering fields. To record a phase image, forward scattered light from a thin object is collected in the back Fourier plane of a collection optic. Placing a narrow slit in the back Fourier plane allows the complex spatial frequency spectrum of the object amplitude transmission to be recorded in time. At each time point, the spatial frequency value corresponding to the difference in transverse spatial frequency of the illumination beams is recorded. Simultaneously, the interference of the illumination beams in the object imparts a spatial frequency pattern on the fluorescent molecule excitation and the spatial frequency of the object’s fluorescent concentration is recorded at each time step. This single-pixel imaging method allows for simultaneous acquisition of the object phase and fluorescent images by collecting spatial frequency projections in time.
Patrick A. Stockton, "Simultaneous fluorescent and quantitative phase and imaging through spatial frequency projections (Conference Presentation)," Proc. SPIE 10074, Quantitative Phase Imaging III, 100740R (Presented at SPIE BiOS: January 30, 2017; Published: 24 April 2017); https://doi.org/10.1117/12.2253482.5380600135001.
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