Real-time simultaneous refractive index and thickness mapping of sub-cellular biology at the diffraction limit

Commun Biol. 2024 Feb 6;7(1):154. doi: 10.1038/s42003-024-05839-w.

Abstract

Mapping the cellular refractive index (RI) is a central task for research involving the composition of microorganisms and the development of models providing automated medical screenings with accuracy beyond 95%. These models require significantly enhancing the state-of-the-art RI mapping capabilities to provide large amounts of accurate RI data at high throughput. Here, we present a machine-learning-based technique that obtains a biological specimen's real-time RI and thickness maps from a single image acquired with a conventional color camera. This technology leverages a suitably engineered nanostructured membrane that stretches a biological analyte over its surface and absorbs transmitted light, generating complex reflection spectra from each sample point. The technique does not need pre-existing sample knowledge. It achieves 10-4 RI sensitivity and sub-nanometer thickness resolution on diffraction-limited spatial areas. We illustrate practical application by performing sub-cellular segmentation of HCT-116 colorectal cancer cells, obtaining complete three-dimensional reconstruction of the cellular regions with a characteristic length of 30 μm. These results can facilitate the development of real-time label-free technologies for biomedical studies on microscopic multicellular dynamics.

MeSH terms

  • HCT116 Cells
  • Humans
  • Refractometry*