Abstract
AbstractWe present a new label-free three-dimensional (3D) microscopy technique, termed transport of intensity diffraction tomography with non-interferometric synthetic aperture (TIDT-NSA). Without resorting to interferometric detection, TIDT-NSA retrieves the 3D refractive index (RI) distribution of biological specimens from 3D intensity-only measurements at various illumination angles, allowing incoherent-diffraction-limited quantitative 3D phase-contrast imaging. The unique combination of z-scanning the sample with illumination angle diversity in TIDT-NSA provides strong defocus phase contrast and better optical sectioning capabilities suitable for high-resolution tomography of thick biological samples. Based on an off-the-shelf bright-field microscope with a programmable light-emitting-diode (LED) illumination source, TIDT-NSA achieves an imaging resolution of 206 nm laterally and 520 nm axially with a high-NA oil immersion objective. We validate the 3D RI tomographic imaging performance on various unlabeled fixed and live samples, including human breast cancer cell lines MCF-7, human hepatocyte carcinoma cell lines HepG2, mouse macrophage cell lines RAW 264.7, Caenorhabditis elegans (C. elegans), and live Henrietta Lacks (HeLa) cells. These results establish TIDT-NSA as a new non-interferometric approach to optical diffraction tomography and 3D label-free microscopy, permitting quantitative characterization of cell morphology and time-dependent subcellular changes for widespread biological and medical applications.
Publisher
Springer Science and Business Media LLC
Subject
Atomic and Molecular Physics, and Optics,Electronic, Optical and Magnetic Materials
Reference69 articles.
1. Mertz, J. Introduction to Optical Microscopy (Cambridge University Press, 2019).
2. Rost, F. W. D. Fluorescence Microscopy (Cambridge University Press, 1992).
3. Lichtman, J. W. & Conchello, J. A. Fluorescence microscopy. Nat. Methods 2, 910–919 (2005).
4. Webb, R. H. Confocal optical microscopy. Rep. Prog. Phys. 59, 427–471 (1996).
5. Sheppard, C. J. R., Hamilton, D. K. & Matthews, H. J. Confocal interference microscopy. Proceedings of SPIE 1028, Scanning Imaging. (SPIE,Hamburg, Germany, 1988).
Cited by
99 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献