Multiscale label-free volumetric holographic histopathology of thick-tissue slides with subcellular resolution

Abstract

Histopathology relies upon the staining and sectioning of biological tissues, which can be laborious and may cause artefacts and distort tissues. Here, we demonstrate label-free volumetric imaging of thick-tissue slides, exploiting refractive index distributions as intrinsic imaging contrast. The present method systematically exploits label-free quantitative phase imaging techniques, volumetric reconstruction of intrinsic refractive index distributions in tissues, and numerical algorithms for the seamless stitching of multiple 3D tomograms and for reducing scattering-induced image distortion. We demonstrate demonstrated label-free volumetric imaging of thick tissues with the field of view of 2 mm x 1.75 mm x 0.2 mm with a spatial resolution of 170 nm x 170 nm x 1200 nm. The number of optical modes, calculated as the reconstructed volume divided by the size of the point spread function, was approximately 20 Giga voxels. We have also demonstrated that different tumour types, and a variety of precursor lesions and pathologies can be visualized with the present method. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC="FIGDIR/small/205633v1_ufig1.gif" ALT="Figure 1"> View larger version (98K): org.highwire.dtl.DTLVardef@dcf15corg.highwire.dtl.DTLVardef@16a05d3org.highwire.dtl.DTLVardef@1a48860org.highwire.dtl.DTLVardef@f57430_HPS_FORMAT_FIGEXP M_FIG C_FIG