Multiple-scattering suppressive refractive index tomography for the label-free quantitative assessment of multicellular spheroids.

Refractive index (RI) tomography is a quantitative tomographic technique used to visualize the intrinsic contrast of unlabeled biological samples. Conventional RI reconstruction algorithms are based on weak-scattering approximation, such as the Born or Rytov approximation. Although these linear algorithms are computationally efficient, they are invalid when the fields are strongly distorted by multiple scattering (MS) of specimens. Herein, we propose an approach to reconstruct the RI distributions of MS objects even under weak-scattering approximation using an MS-suppressive operation. The operation converts the distorted fields into MS-suppressed fields, where weak-scattering approximation is applicable. Using this approach, we reconstructed a whole multicellular spheroid and successfully visualized its internal subcellular structures. Our work facilitates the realization of RI tomography of MS specimens and label-free quantitative analysis of 3D multicellular specimens.