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Optical computed tomography for spatially isotropic four-dimensional imaging of live single cells. Sci Adv 2017 12;3(12):e1602580

Date

12/12/2017

Pubmed ID

29226240

Pubmed Central ID

PMC5721812

DOI

10.1126/sciadv.1602580

Scopus ID

2-s2.0-85041821749   4 Citations

Abstract

Quantitative three-dimensional (3D) computed tomography (CT) imaging of living single cells enables orientation-independent morphometric analysis of the intricacies of cellular physiology. Since its invention, x-ray CT has become indispensable in the clinic for diagnostic and prognostic purposes due to its quantitative absorption-based imaging in true 3D that allows objects of interest to be viewed and measured from any orientation. However, x-ray CT has not been useful at the level of single cells because there is insufficient contrast to form an image. Recently, optical CT has been developed successfully for fixed cells, but this technology called Cell-CT is incompatible with live-cell imaging due to the use of stains, such as hematoxylin, that are not compatible with cell viability. We present a novel development of optical CT for quantitative, multispectral functional 4D (three spatial + one spectral dimension) imaging of living single cells. The method applied to immune system cells offers truly isotropic 3D spatial resolution and enables time-resolved imaging studies of cells suspended in aqueous medium. Using live-cell optical CT, we found a heterogeneous response to mitochondrial fission inhibition in mouse macrophages and differential basal remodeling of small (0.1 to 1 fl) and large (1 to 20 fl) nuclear and mitochondrial structures on a 20- to 30-s time scale in human myelogenous leukemia cells. Because of its robust 3D measurement capabilities, live-cell optical CT represents a powerful new tool in the biomedical research field.

Author List

Kelbauskas L, Shetty R, Cao B, Wang KC, Smith D, Wang H, Chao SH, Gangaraju S, Ashcroft B, Kritzer M, Glenn H, Johnson RH, Meldrum DR

Author

Roger H. Johnson PhD Associate Professor in the Biophysics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Cell Nucleus
Equipment Design
Four-Dimensional Computed Tomography
Humans
K562 Cells
Mitochondria
Reproducibility of Results
Single-Cell Analysis
Tomography, Optical