Abstract
We explain how to concentrate light simultaneously at multiple selected volumetric positions by means of a 4D illumination light field. First, to select target objects, a 4D imaging light field is captured. A light field mask is then computed automatically for this selection to avoid illumination of the remaining areas. With one-photon illumination, simultaneous generation of complex volumetric light patterns becomes possible. As a full light-field can be captured and projected simultaneously at the desired exposure and excitation times, short readout and lighting durations are supported.
Highlights
We explain how to concentrate light simultaneously at multiple selected volumetric positions by means of a 4D illumination light field
Precise controllable excitation is another requirement in recent microscopy techniques
Multiple simultaneous beams can be generated with spatial light modulators (SLMs), such as digital micromirror devices[9], or phase-modulators (PMs), such as liquid crystal PMs used for holographic projection[10]
Summary
According to Sparrow’s criterion, the smallest resolvable spot for wavelength λ, numerical aperture NA, and magnification M is o equals the microlens. While the lateral resolution of a light-field microscope can be increased mainly by downscaling the microlens pitch of the MLA, the axial resolution is enhanced by reducing the pixel pitch of the DMD (down to the diffraction limit set by o), increasing the NA of the objective, and reducing the excitation wavelength. Future experiments on actual organic tissue will provide further insights
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