Imaging thick samples with optical tomography

TL;DR

This paper advances optical diffraction tomography (ODT) by enabling imaging of multi-cell clusters in thicker samples, assessing algorithm performance, and demonstrating a new multi-layer imaging strategy.

Contribution

It introduces a novel approach for imaging multi-layer cell clusters with ODT, expanding its application to thicker biological samples.

Findings

ODT can be adapted for thicker, multi-layer samples.

Sample thickness impacts reconstruction quality depending on the algorithm.

A new imaging strategy successfully visualizes multi-cell clusters.

Abstract

Optical diffraction tomography (ODT), initially described in the seminal paper of Emil Wolf [Opt. Comm., 1(4), 153-156 (1969)], has received renewed attention recently. The vast majority of ODT experiments reported to date have been 3D images of single cells or beads demonstrating excellent image quality and sectioning capability. The sample thickness in these experiments is 10 to 20 microns and the optical thickness is generally well below 2{\pi}. In this paper, we explore ODT when the sample consists of multiple layers of cells. We assess experimentally the impact of sample thickness for the different ODT reconstruction algorithms and we describe a strategy that allows us to image, for the first time, multi-cell clusters.