Stain-Free Quantification of Chromosomes in Live Cells Using Regularized Tomographic Phase Microscopy

Sung, Yongjin; Choi, Wonshik; Lue, Niyom; Dasari, Ramachandra R.; Yaqoob, Zahid

doi:10.1371/journal.pone.0049502

Cited by 103 publications

(87 citation statements)

References 40 publications

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“…Our method collects images in a broad range of wavelength, each of which contains both material dispersion information as well as structural information of the sample. Extracting material dispersion of the cellular organelles, which is left for a future study, could provide rich molecular-specific information, e.g., relative amount of nucleic acids and proteins, at a subcellular level [4].…”

Section: Discussionmentioning

confidence: 99%

“…The refractive index is the source of image contrast in DHM, which can be related to the average concentration of non-aqueous contents within the specimen, the socalled dry mass [1][2][3]. Tomographic interrogation of living biological specimens using DHM can thereby provide the mass of cellular organelles [4] as well as three-dimensional morphology of the subcellular structures within cells and small organisms [5]. There are other label-free approaches [6,7] that rely on non-linear light-matter interactions to provide rich molecular-specific information.…”

Section: Introductionmentioning

confidence: 99%

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Scanning color optical tomography (SCOT)

Hosseini¹,

Sung²,

Choi³

et al. 2015

Opt. Express

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Abstract:We have developed an interferometric optical microscope that provides three-dimensional refractive index map of a specimen by scanning the color of three illumination beams. Our design of the interferometer allows for simultaneous measurement of the scattered fields (both amplitude and phase) of such a complex input beam. By obviating the need for mechanical scanning of the illumination beam or detection objective lens; the proposed method can increase the speed of the optical tomography by orders of magnitude. We demonstrate our method using polystyrene beads of known refractive index value and live cells. 349-393 (1988). 12. M. Reed Teague, "Deterministic phase retrieval: a Green's function solution," J. Opt. Soc. Am. 73(11), 1434-1441 (1983). 13. F. Charrière, A. Marian, F. Montfort, J. Kuehn, T. Colomb, E. Cuche, P. Marquet, and C. Depeursinge, "Cell refractive index tomography by digital holographic microscopy," Opt. Lett. 31(2), 178-180 (2006). 14. V. Lauer, "New approach to optical diffraction tomography yielding a vector equation of diffraction tomography and a novel tomographic microscope," J. Microsc. 205(2), 165-176 (2002 Opt. Commun. 1(7), 323-328 (1970). 28. G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, "Diffraction phase microscopy for quantifying cell structure and dynamics," Opt. Lett. 31(6), 775-777 (2006). (McGraw Hill Professional, 2011). 30. E. Wolf, "Three-dimensional structure determination of semi-transparent objects from holographic data," Opt. G. Popescu, Quantitative Phase Imaging of Cells and TissuesCommun. 1(4), 153-156 (1969). 31. A. J. Devaney, "Inverse-scattering theory within the Rytov approximation," Opt. Lett. 6(8), 374-376 (1981).

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Scanning color optical tomography (SCOT)

Hosseini¹,

Sung²,

Choi³

et al. 2015

Opt. Express

Self Cite

View full text Add to dashboard Cite

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“…Although the number of applications utilizing QPI [5][6][7][8][9][10][11][12][13] is rising steadily, QPI, as a technology, is under-utilized in comparison with non-quantitative phase contrast. One possible explanation for this is that modalities such as PC, DIC, and fluorescence microscopy may be integrated into a single optical system with excellent imaging properties (specklefree, uniform) due to the use of partially coherent (both temporally and spatially, usually Köhler) illumination from filament light sources whereas QPI conventionally requires coherent illumination which is not ideal for imaging with speckle-interference and phase jitter as intrinsic noise sources [4].…”

Section: Introductionmentioning

confidence: 99%

Bright-field quantitative phase microscopy (BFQPM) for accurate phase imaging using conventional microscopy hardware

Jenkins¹,

Gaylord²

2015

SPIE Proceedings

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Most quantitative phase microscopy methods require the use of custom-built or modified microscopic configurations which are not typically available to most bio/pathologists. There are, however, phase retrieval algorithms which utilize defocused bright-field images as input data and are therefore implementable in existing laboratory environments. Among these, deterministic methods such as those based on inverting the transport-of-intensity equation (TIE) or a phase contrast transfer function (PCTF) are particularly attractive due to their compatibility with Köhler illuminated systems and numerical simplicity. Recently, a new method has been proposed, called multi-filter phase imaging with partially coherent light (MFPI-PC), which alleviates the inherent noise/resolution trade-off in solving the TIE by utilizing a large number of defocused bright-field images spaced equally about the focal plane. Despite greatly improving the state-ofthe-art, the method has many shortcomings including the impracticality of high-speed acquisition, inefficient sampling, and attenuated response at high frequencies due to aperture effects. In this report, we present a new method, called bright-field quantitative phase microscopy (BFQPM), which efficiently utilizes a small number of defocused bright-field images and recovers frequencies out to the partially coherent diffraction limit. The method is based on a noiseminimized inversion of a PCTF derived for each finite defocus distance. We present simulation results which indicate nanoscale optical path length sensitivity and improved performance over MFPI-PC. We also provide experimental results imaging live bovine mesenchymal stem cells at sub-second temporal resolution. In all, BFQPM enables fast and accurate phase imaging with unprecedented spatial resolution using widely available bright-field microscopy hardware.

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“…을 활용하거나 [4,5] 자외선 영역의 빛을 사용하여 세포의 핵을 구 분하고, [6] 세포 내 타깃 단백질에 선택적으로 높은 굴절률을 가 지는 금나노입자(gold nanoparticle)를 부착함으로써 세포 내 기 관들을 구분하려는 연구 [7] 가 활발히 진행되고 있다. Reproduced from Refs.…”

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