Abstract
We report the development of a line-scan focal modulation microscope (LSFMM) that is capable of high-speed image acquisition ( > 40 ?? fps ) with uncompromised optical sectioning capability. The improved background rejection and axial resolution of this imaging modality, enabled by focal modulation, are quantified with three-dimensional imaging data obtained from fluorescent beads. The signal-to-background ratio for the LSFMM system is one- to two-orders of magnitude higher than that for line-scanning confocal systems when imaging deep (up to 100 ?m) into a turbid medium of optical properties similar to biological tissues. The imaging performance of LSFMM, in terms of both spatial and temporal resolutions, is further demonstrated with in vivo imaging experiments with live zebrafish larvae.
Highlights
We report the development of a line-scan focal modulation microscope (LSFMM) that is capable of highspeed image acquisition (>40 fps) with uncompromised optical sectioning capability
The superiority of Focal modulation microscopy (FMM) over confocal systems in terms of signal-to-background ratio (SBR) and penetration depth is evident from the literature.[14,17,18]
LSFMM speeds up the image acquisition process by scanning a line focus in one-dimension (1-D) only
Summary
We report the development of a line-scan focal modulation microscope (LSFMM) that is capable of highspeed image acquisition (>40 fps) with uncompromised optical sectioning capability. Focal modulation microscopy (FMM) is an emerging microscopy technique that has the ability to image up to 700 μm in depth in optically thick tissues.[14,15,16] It employs a spatiotemporal phase modulator in the excitation light path to modulate the phase of the excitation beam spatially and temporally (at a fixed frequency). LSFMM speeds up the image acquisition process by scanning a line focus in one-dimension (1-D) only.
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