Size homeostasis in adherent cells studied by synthetic phase microscopy

Sung, Yongjin; Tzur, Amit; Oh, Seungeun; Choi, Wonshik; Li, Victor C.; Dasari, Ramachandra R.; Yaqoob, Zahid; Kirschner, Marc W.

doi:10.1073/pnas.1315290110

Cited by 98 publications

(106 citation statements)

References 48 publications

(66 reference statements)

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“…Important to note is that the different setups come with slightly different benefits and limitations, making them inherently well suited to a variety of purposes: High speed acquisition for high temporal resolution of flow imaging [70], in situ imaging of growing adherent cells [21,37,56], live cell tomography [71,72], or incubator monitoring [30,73].…”

Section: Technologymentioning

confidence: 99%

“…The ability to measure cell count was validated for the first time in 2008 by Mölder et al [21], using human breast and prostate cancer cell lines, as well as a mouse fibroblast cell line. Measurement of cell growth has been well documented for both adherent and non-adherent cells [8,9,15,[25][26][27][28]47,49,56,59,61]. By segmentation of cellular outlines, morphology and motility can also be studied [7,50,51,57,80].…”

Section: Principles Of Qpimentioning

confidence: 99%

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Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

2018

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Abstract:To understand complex biological processes, scientists must gain insight into the function of individual living cells. In contrast to the imaging of fixed cells, where a single snapshot of the cell's life is retrieved, live-cell imaging allows investigation of the dynamic processes underlying the function and morphology of cells. Label-free imaging of living cells is advantageous since it is used without fluorescent probes and maintains an appropriate environment for cellular behavior, otherwise leading to phototoxicity and photo bleaching. Quantitative phase imaging (QPI) is an ideal method for studying live cell dynamics by providing data from noninvasive monitoring over arbitrary time scales. The effect of drugs on migration, proliferation, and apoptosis of cancer cells are emerging fields suitable for QPI analysis. In this review, we provide a current insight into QPI applied to cancer research.

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Section: Technologymentioning

confidence: 99%

Section: Principles Of Qpimentioning

confidence: 99%

Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

2018

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“…Thus, cell size is proportional to growth rate, which means that slow growing cells can be nearly half the size of rapidly growing cells (Johnston et al 1977;Fantes and Nurse 1977). Conversely, at least in some cases cell size influences growth rate so that large cells grow faster than small cells (Tzur et al 2009;Sung et al 2013;Schmoller et al 2015;Leitao and Kellogg 2017). Together, these observations show that growth rate is matched to nutrient availability, cell size is matched to growth rate, and growth rate is matched to cell size.…”

Section: Introductionmentioning

confidence: 95%

Modulation of TORC2 Signaling by a Conserved Lkb1 Signaling Axis in Budding Yeast

et al. 2018

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Nutrient availability, growth rate and cell size are closely linked. For example, in budding yeast, the rate of cell growth is proportional to nutrient availability, cell size is proportional to growth rate, and growth rate is proportional to cell size. Thus, cells grow slowly in poor nutrients and are nearly half the size of cells growing in rich nutrients. Moreover, large cells grow faster than small cells. A signaling network that surrounds Tor kinase complex 2 (TORC2) plays an important role in enforcing these proportional relationships. Cells that lack components of the TORC2 network fail to modulate their growth rate or size in response to changes in nutrient availability. Here, we show that budding yeast homologs of the Lkb1 tumor suppressor kinase are required for normal modulation of TORC2 signaling in response to changes in carbon source. Lkb1 kinases activate Snf1/AMPK to initiate transcription of genes required for utilization of poor carbon sources. However, Lkb1 influences TORC2 signaling via a novel pathway that is independent of Snf1/AMPK. Of the three Lkb1 homologs in budding yeast, Elm1 plays the most important role in modulating TORC2. Elm1 activates a pair of related kinases called Gin4 and Hsl1. Previous work found that loss of Gin4 and Hsl1 causes cells to undergo unrestrained growth during a prolonged mitotic arrest, which suggests that they play a role in linking cell cycle progression to cell growth. We found that Gin4 and Hsl1 also control the TORC2 network. In addition, Gin4 and Hsl1 are themselves influenced by signals from the TORC2 network, consistent with previous work showing that the TORC2 network constitutes a feedback loop. Together, the data suggest a model in which the TORC2 network sets growth rate in response to carbon source, while also relaying signals via Gin4 and Hsl1 that set the critical amount of growth required for cell cycle progression. This kind of close linkage between control of cell growth and size would suggest a simple mechanistic explanation for the proportional relationship between cell size and growth rate.3

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“…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].…”

Section: Introductionmentioning

confidence: 99%

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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Size homeostasis in adherent cells studied by synthetic phase microscopy

Cited by 98 publications

References 48 publications

Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

Modulation of TORC2 Signaling by a Conserved Lkb1 Signaling Axis in Budding Yeast

Scanning color optical tomography (SCOT)

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