We demonstrate "depth of field multiplexing" by a high resolution spatial light modulator (SLM) in a Fourier plane in the imaging path of a standard microscope. This approach provides simultaneous imaging of different focal planes in a sample with only a single camera exposure. The phase mask on the SLM corresponds to a set of superposed multi-focal off-axis Fresnel lenses, which sharply image different focal planes of the object to non-overlapping adjacent sections of the camera chip. Depth of field multiplexing allows to record motion in a three dimensional sample volume in real-time, which is exemplarily demonstrated for cytoplasmic streaming in plant cells and rapidly swimming protozoa.
We describe the implementation of quantitative Differential Interference Contrast (DIC) Microscopy using a spatial light modulator (SLM) as a flexible Fourier filter in the optical path. The experimental arrangement allows for the all-electronic acquisition of multiple phase shifted DIC-images at video rates which are analyzed to yield the optical path length variation of the sample. The resolution of the technique is analyzed by retrieving the phase profiles of polystyrene spheres in immersion oil, and the method is then applied for quantitative imaging of biological samples. By reprogramming the diffractive structure displayed at the SLM it is possible to record the whole set of phase shifted DIC images simultaneously in different areas of the same camera chip. This allows for quantitative snap-shot imaging of a sample, which has applications for the investigation of dynamic processes.
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