Differential structured illumination microscopy (dSIM) for high-throughput 3D biological imaging

High-throughput 3D imaging of biological structures and their dynamics has become increasingly important for biological applications including immune response to infection, cancer growth, and cellular metabolism. Recent computational imaging modalities coupling novel acquisition strategies with advanced computation techniques have successfully captured such dynamics, but their use of intensive computation can incur multi-day or weeklong reconstruction timescales preventing biologists from directly analyzing their sample’s behavior. Here, we developed differential structured illumination microscopy (dSIM) for capturing 3D biological sample dynamics at near-isotropic 1μm 3D resolution over a 1mm2 field-of-view at 10Hz volume rates. This modality applies plane waves and grating-based illuminations in a differential scheme to encode high-angle, traditionally nonlinear “darkfield” illuminations into linear intensity measurements for efficient 3D object reconstruction with linear inverse scattering models. Capturing this darkfield information enables 4.5X better lateral resolution over the coherent imaging bandwidth. We illustrate this technique on cell cultures and thin tissue slices.