Super-resolution three-dimensional fluorescence and optical diffraction tomography of live cells using structured illumination generated by a digital micromirror device

Kyoohyun Kim,Yongkeun Park,Seungwoo Shin,Doyeon Kim

doi:10.1038/s41598-018-27399-w

Kyoohyun Kim, Yongkeun Park + Show 2 more

Open Access

https://doi.org/10.1038/s41598-018-27399-w

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Abstract

We present a multimodal approach for measuring the three-dimensional (3D) refractive index (RI) and fluorescence distributions of live cells by combining optical diffraction tomography (ODT) and 3D structured illumination microscopy (SIM). A digital micromirror device is utilized to generate structured illumination patterns for both ODT and SIM, which enables fast and stable measurements. To verify its feasibility and applicability, the proposed method is used to measure the 3D RI distribution and 3D fluorescence image of various samples, including a cluster of fluorescent beads, and the time-lapse 3D RI dynamics of fluorescent beads inside a HeLa cell, from which the trajectory of the beads in the HeLa cell is analyzed using spatiotemporal correlations.

Highlights

Fluorescence distributions of live cells by combining optical diffraction tomography (ODT) and 3D structured illumination microscopy (SIM)
To minimize photobleaching and phototoxicity that occur in fluorescence imaging of live cells, the two operating modes are separate utilizing dispersion property of a digital micromirror device (DMD) and two lasers with different wavelengths
By selectively blocking one of the laser beams, the present setup alternated between the ODT mode and the 3D SIM

Summary

Introduction

Fluorescence distributions of live cells by combining optical diffraction tomography (ODT) and 3D structured illumination microscopy (SIM). Optical diffraction tomography (ODT) or 3D quantitative phase imaging (QPI) techniques have emerged as methods for label-free quantitative imaging of 3D refractive index (RI) distributions of biological samples[15,16,17,18,19,20]. The. RI distribution of live cells has provided structural and biochemical information about biological samples in studies in the fields of cell biology[23,24], microbiology[25], hematology[26], infectious diseases[27], plant biology[28], and biophysics[29,30]. The RI generally provides limited molecular specificity[31], except for certain materials with distinct RI values such as lipids[32,33] and metallic particles[34,35]

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