Abstract
To extend the imaging depth of high-resolution optical microscopy, various gating operations—confocal, coherence, and polarization gating—have been devised to filter out the multiply scattered wave. However, the imaging depth is still limited by the multiply scattered wave that bypasses the existing gating operations. Here, we present a space gating method, whose mechanism is independent of the existing methods and yet effective enough to complement them. Specifically, we reconstruct an image only using the ballistic wave that is acousto-optically modulated at the object plane. The space gating suppresses the multiply scattered wave by 10–100 times in a highly scattering medium, and thus enables visualization of the skeletal muscle fibers in whole-body zebrafish at 30 days post fertilization. The space gating will be an important addition to optical-resolution microscopy for achieving the ultimate imaging depth set by the detection limit of ballistic wave.
Highlights
To extend the imaging depth of high-resolution optical microscopy, various gating operations —confocal, coherence, and polarization gating—have been devised to filter out the multiply scattered wave
The space gating aims to selectively suppress the multiply scattered wave based on the fact that it is spatially spread over the wide extent on the object plane, in contrast to the ballistic wave which is tightly confined at the confocal point
The space gating is implemented by setting a spatial window RSG around the confocal point on the object plane in such a way that only the wave transmitted through the gating window contributes to the detected field
Summary
To extend the imaging depth of high-resolution optical microscopy, various gating operations —confocal, coherence, and polarization gating—have been devised to filter out the multiply scattered wave. Various adaptive optics approaches have been proposed to maintain the effectiveness of gating operations in spite of sampleinduced aberration[14,15,16,17] Even with these substantial advances, the imaging depth of high-resolution optical microscopy has not yet reached the detection limit set by the dynamic range of state-of-the-art sensor technology. Unlike confocal or time gating, space gating is directly applied at the object plane inside the scattering medium to reject the multiply scattered wave whose optical path spreads beyond the extent of the ultrasound focus. It can remove the multiply scattered wave, which cannot be filtered out by the existing gating operations. It represents an important step toward reaching the fundamental depth limit of diffraction-limited imaging relying on ballistic waves, and opens new possibilities for label-free imaging of biological cells through scattering tissues
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