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Artikel ; Online: Fluorescent biosensor imaging meets deterministic mathematical modelling: quantitative investigation of signalling compartmentalization.

Posner, Clara / Mehta, Sohum / Zhang, Jin

2023 Band 601, Heft 19, Seite(n) 4227–4241

Abstract: Cells execute specific responses to diverse environmental cues by encoding information in distinctly compartmentalized biochemical signalling reactions. Genetically encoded fluorescent biosensors enable the spatial and temporal monitoring of signalling ... ...

Abstract	Cells execute specific responses to diverse environmental cues by encoding information in distinctly compartmentalized biochemical signalling reactions. Genetically encoded fluorescent biosensors enable the spatial and temporal monitoring of signalling events in live cells. Temporal and spatiotemporal computational models can be used to interpret biosensor experiments in complex biochemical networks and to explore hypotheses that are difficult to test experimentally. In this review, we first provide brief discussions of the experimental toolkit of fluorescent biosensors as well as computational basics with a focus on temporal and spatiotemporal deterministic models. We then describe how we used this combined approach to identify and investigate a protein kinase A (PKA) - cAMP - Ca
Mesh-Begriff(e)	Animals ; Mice ; Signal Transduction ; Cyclic AMP-Dependent Protein Kinases/metabolism ; Diagnostic Imaging ; Cell Membrane/metabolism ; Biosensing Techniques/methods ; Fluorescence Resonance Energy Transfer/methods
Chemische Substanzen	Cyclic AMP-Dependent Protein Kinases (EC 2.7.11.11)
Sprache	Englisch
Erscheinungsdatum	2023-09-25
Erscheinungsland	England
Dokumenttyp	Review ; Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	3115-x
ISSN	1469-7793 ; 0022-3751
ISSN (online)	1469-7793
ISSN	0022-3751
DOI	10.1113/JP282696
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Author Correction: A rationally enhanced red fluorescent protein expands the utility of FRET biosensors.

Mo, Gary C H / Posner, Clara / Rodriguez, Erik A / Sun, Tengqian / Zhang, Jin

Nature communications

2024 Band 15, Heft 1, Seite(n) 771

Sprache	Englisch
Erscheinungsdatum	2024-01-26
Erscheinungsland	England
Dokumenttyp	Published Erratum
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-024-45330-y
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: GPCR Signaling Measurement and Drug Profiling with an Automated Live-Cell Microscopy System.

Greenwald, Eric / Posner, Clara / Bharath, Ananya / Lyons, Anne / Salmerón, Cristina / Sriram, Krishna / Wiley, Shu Z / Insel, Paul A / Zhang, Jin

ACS sensors

2023 Band 8, Heft 1, Seite(n) 19–27

Abstract: A major limitation of time-lapse microscopy combined with fluorescent biosensors, a powerful tool for quantifying spatiotemporal dynamics of signaling in single living cells, is low-experimental throughput. To overcome this limitation, we created a ... ...

Abstract	A major limitation of time-lapse microscopy combined with fluorescent biosensors, a powerful tool for quantifying spatiotemporal dynamics of signaling in single living cells, is low-experimental throughput. To overcome this limitation, we created a highly customizable, MATLAB-based platform: flexible automated liquid-handling combined microscope (FALCOscope) that coordinates an OpenTrons liquid handler and a fluorescence microscope to automate drug treatments, fluorescence imaging, and single-cell analysis. To test the feasibility of the FALCOscope, we quantified G protein-coupled receptor (GPCR)-stimulated Protein Kinase A activity and cAMP responses to GPCR agonists and antagonists. We also characterized cAMP dynamics induced by GPR68/OGR1, a proton-sensing GPCR, in response to variable extracellular pH values. GPR68-induced cAMP responses were more transient in acidic than neutral pH values, suggesting a pH-dependence for signal attenuation. Ogerin, a GPR68 positive allosteric modulator, enhanced cAMP response most strongly at pH 7.0 and sustained cAMP response for acidic pH values, thereby demonstrating the capability of the FALCOscope to capture allosteric modulation. At a high concentration, ogerin increased cAMP signaling independent of GPR68, likely via phosphodiesterase inhibition. The FALCOscope system thus enables enhanced throughput single-cell dynamic measurements and is a versatile system for interrogating spatiotemporal regulation of signaling molecules in living cells and for drug profiling and screening.
Mesh-Begriff(e)	Benzyl Alcohols/pharmacology ; Microscopy, Fluorescence ; Signal Transduction ; Triazines ; Receptors, G-Protein-Coupled/metabolism
Chemische Substanzen	Benzyl Alcohols ; ogerin ; Triazines ; Receptors, G-Protein-Coupled
Sprache	Englisch
Erscheinungsdatum	2023-01-05
Erscheinungsland	United States
Dokumenttyp	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ISSN	2379-3694
ISSN (online)	2379-3694
DOI	10.1021/acssensors.2c01341
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Imaging of Cell Morphology Changes via Metamaterial-Assisted Photobleaching Microscopy.

Lee, Yeon Ui / Posner, Clara / Zhao, Junxiang / Zhang, Jin / Liu, Zhaowei

Nano letters

2021 Band 21, Heft 4, Seite(n) 1716–1721

Abstract: Determining the axial position of an emitter with nanoscale precision is critical to a fundamental imaging methodology. While there are many advanced optical techniques being applied to high-resolution imaging, high-axial-resolution topography imaging of ...

Abstract	Determining the axial position of an emitter with nanoscale precision is critical to a fundamental imaging methodology. While there are many advanced optical techniques being applied to high-resolution imaging, high-axial-resolution topography imaging of living cells is particularly challenging. Here, we present an application of metamaterial-assisted photobleaching microscopy (MAPM) with high-axial resolution to characterize morphological properties of living cells. Quantitative imaging of changes in the morphology of live cells is obtained by topographic and statistical analysis. The time-lapse topography image using the metamaterial-induced photostability provides information about growth factor induced changes in the cell morphology with high-axial resolution.
Mesh-Begriff(e)	Microscopy, Fluorescence ; Photobleaching
Sprache	Englisch
Erscheinungsdatum	2021-02-12
Erscheinungsland	United States
Dokumenttyp	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	1530-6992
ISSN (online)	1530-6992
DOI	10.1021/acs.nanolett.0c04529
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Fourier Optical Spin Splitting Microscopy.

Zhou, Junxiao / Wu, Qianyi / Zhao, Junxiang / Posner, Clara / Lei, Ming / Chen, Guanghao / Zhang, Jin / Liu, Zhaowei

Physical review letters

2022 Band 129, Heft 2, Seite(n) 20801

Abstract: In this Letter, we propose a new quantitative phase imaging methodology named Fourier optical spin splitting microscopy (FOSSM). FOSSM relies on a metasurface located at the Fourier plane of a polarized microscope to separate the object image into two ... ...

Abstract	In this Letter, we propose a new quantitative phase imaging methodology named Fourier optical spin splitting microscopy (FOSSM). FOSSM relies on a metasurface located at the Fourier plane of a polarized microscope to separate the object image into two replicas of opposite circularly polarized states. The bias retardation between the two replicas is tuned by translating the metasurface or rotating the analyzer. Combined with a polarized camera, FOSSM can easily achieve single-shot quantitative phase gradient imaging, which greatly reduces the complexity of current phase microscope setups, paving the way for the next generation high-speed real-time multifunctional microscopy.
Mesh-Begriff(e)	Microscopy/methods
Sprache	Englisch
Erscheinungsdatum	2022-07-19
Erscheinungsland	United States
Dokumenttyp	Journal Article
ZDB-ID	208853-8
ISSN	1079-7114 ; 0031-9007
ISSN (online)	1079-7114
ISSN	0031-9007
DOI	10.1103/PhysRevLett.129.020801
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: A rationally enhanced red fluorescent protein expands the utility of FRET biosensors.

Mo, Gary C H / Posner, Clara / Rodriguez, Erik A / Sun, Tengqian / Zhang, Jin

Nature communications

2020 Band 11, Heft 1, Seite(n) 1848

Abstract: Genetically encoded Förster Resonance Energy Transfer (FRET)-based biosensors are powerful tools to illuminate spatiotemporal regulation of cell signaling in living cells, but the utility of the red spectrum for biosensing was limited due to a lack of ... ...

Abstract	Genetically encoded Förster Resonance Energy Transfer (FRET)-based biosensors are powerful tools to illuminate spatiotemporal regulation of cell signaling in living cells, but the utility of the red spectrum for biosensing was limited due to a lack of bright and stable red fluorescent proteins. Here, we rationally improve the photophysical characteristics of the coral-derived fluorescent protein TagRFP-T. We show that a new single-residue mutant, super-TagRFP (stagRFP) has nearly twice the molecular brightness of TagRFP-T and negligible photoactivation. stagRFP facilitates significant improvements on multiple green-red biosensors as a FRET acceptor and is an efficient FRET donor that supports red/far-red FRET biosensing. Capitalizing on the ability of stagRFP to couple with multiple FRET partners, we develop a novel multiplex method to examine the confluence of signaling activities from three kinases simultaneously in single living cells, providing evidence for a role of Src family kinases in regulating growth factor induced Akt and ERK activities.
Mesh-Begriff(e)	Fluorescence Resonance Energy Transfer ; Humans ; Luminescent Proteins/chemistry ; Mutagenesis/genetics ; Mutagenesis/physiology ; Signal Transduction/genetics ; Signal Transduction/physiology ; Red Fluorescent Protein
Chemische Substanzen	Luminescent Proteins
Sprache	Englisch
Erscheinungsdatum	2020-04-15
Erscheinungsland	England
Dokumenttyp	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-020-15687-x
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Metamaterial assisted illumination nanoscopy via random super-resolution speckles.

Lee, Yeon Ui / Zhao, Junxiang / Ma, Qian / Khorashad, Larousse Khosravi / Posner, Clara / Li, Guangru / Wisna, G Bimananda M / Burns, Zachary / Zhang, Jin / Liu, Zhaowei

Nature communications

2021 Band 12, Heft 1, Seite(n) 1559

Abstract: Structured illumination microscopy (SIM) is one of the most powerful and versatile optical super-resolution techniques. Compared with other super-resolution methods, SIM has shown its unique advantages in wide-field imaging with high temporal resolution ... ...

Abstract	Structured illumination microscopy (SIM) is one of the most powerful and versatile optical super-resolution techniques. Compared with other super-resolution methods, SIM has shown its unique advantages in wide-field imaging with high temporal resolution and low photon damage. However, traditional SIM only has about 2 times spatial resolution improvement compared to the diffraction limit. In this work, we propose and experimentally demonstrate an easily-implemented, low-cost method to extend the resolution of SIM, named speckle metamaterial-assisted illumination nanoscopy (speckle-MAIN). A metamaterial structure is introduced to generate speckle-like sub-diffraction-limit illumination patterns in the near field with improved spatial frequency. Such patterns, similar to traditional SIM, are then used to excite objects on top of the surface. We demonstrate that speckle-MAIN can bring the resolution down to 40 nm and beyond. Speckle-MAIN represents a new route for super-resolution, which may lead to important applications in bio-imaging and surface characterization.
Mesh-Begriff(e)	Imaging, Three-Dimensional/methods ; Microscopy/methods ; Microscopy, Fluorescence/methods
Sprache	Englisch
Erscheinungsdatum	2021-03-10
Erscheinungsland	England
Dokumenttyp	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-021-21835-8
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Organic Hyperbolic Material Assisted Illumination Nanoscopy.

Lee, Yeon Ui / Posner, Clara / Nie, Zhaoyu / Zhao, Junxiang / Li, Shilong / Bopp, Steven Edward / Wisna, Gde Bimananda Mahardika / Ha, Jeongho / Song, Chengyu / Zhang, Jin / Yang, Sui / Zhang, Xiang / Liu, Zhaowei

Advanced science (Weinheim, Baden-Wurttemberg, Germany)

2021 Band 8, Heft 22, Seite(n) e2102230

Abstract: Resolution capability of the linear structured illumination microscopy (SIM) plays a key role in its applications in physics, medicine, biology, and life science. Many advanced methodologies have been developed to extend the resolution of structured ... ...

Abstract	Resolution capability of the linear structured illumination microscopy (SIM) plays a key role in its applications in physics, medicine, biology, and life science. Many advanced methodologies have been developed to extend the resolution of structured illumination by using subdiffraction-limited optical excitation patterns. However, obtaining SIM images with a resolution beyond 40 nm at visible frequency remains as an insurmountable obstacle due to the intrinsic limitation of spatial frequency bandwidth of the involved materials and the complexity of the illumination system. Here, a low-loss natural organic hyperbolic material (OHM) that can support record high spatial-frequency modes beyond 50k
Mesh-Begriff(e)	Equipment Design ; Lighting/instrumentation ; Lighting/methods ; Microscopy, Fluorescence/instrumentation ; Microscopy, Fluorescence/methods ; Nanotechnology/instrumentation ; Nanotechnology/methods
Sprache	Englisch
Erscheinungsdatum	2021-08-26
Erscheinungsland	Germany
Dokumenttyp	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2808093-2
ISSN	2198-3844 ; 2198-3844
ISSN (online)	2198-3844
ISSN	2198-3844
DOI	10.1002/advs.202102230
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Unprecedented Fluorophore Photostability Enabled by Low-Loss Organic Hyperbolic Materials.

Lee, Yeon Ui / Li, Shilong / Bopp, Steven Edward / Zhao, Junxiang / Nie, Zhaoyu / Posner, Clara / Yang, Sui / Zhang, Xiang / Zhang, Jin / Liu, Zhaowei

Advanced materials (Deerfield Beach, Fla.)

2021 Band 33, Heft 9, Seite(n) e2006496

Abstract: The dynamics of photons in fluorescent molecules plays a key role in fluorescence imaging, optical sensing, organic photovoltaics, and displays. Photobleaching is an irreversible photodegradation process of fluorophores, representing a fundamental ... ...

Abstract	The dynamics of photons in fluorescent molecules plays a key role in fluorescence imaging, optical sensing, organic photovoltaics, and displays. Photobleaching is an irreversible photodegradation process of fluorophores, representing a fundamental limitation in relevant optical applications. Chemical reagents are used to suppress the photobleaching rate but with exceptionally high specificity for each type of fluorophore. Here, using organic hyperbolic materials (OHMs), an optical platform to achieve unprecedented fluorophore photostability without any chemical specificity is demonstrated. A more than 500-fold lengthening of the photobleaching lifetime and a 230-fold increase in the total emitted photon counts are observed simultaneously. These exceptional improvements solely come from the low-loss hyperbolic dispersion of OHM films and the large resultant Purcell effect in the visible spectral range. The demonstrated OHM platform may open up a new paradigm in nanophotonics and organic plasmonics for super-resolution imaging and the engineering of light-matter interactions at the nanoscale.
Sprache	Englisch
Erscheinungsdatum	2021-01-27
Erscheinungsland	Germany
Dokumenttyp	Journal Article
ZDB-ID	1474949-X
ISSN	1521-4095 ; 0935-9648
ISSN (online)	1521-4095
ISSN	0935-9648
DOI	10.1002/adma.202006496
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: TAZ is required for lung alveolar epithelial cell differentiation after injury.

Sun, Tianhe / Huang, Zhiyu / Zhang, Hua / Posner, Clara / Jia, Guiquan / Ramalingam, Thirumalai R / Xu, Min / Brightbill, Hans / Egen, Jackson G / Dey, Anwesha / Arron, Joseph R

JCI insight

2019 Band 5

Abstract: The lung is a relatively quiescent organ during homeostasis, but has a remarkable capacity for repair after injury. Alveolar epithelial type I cells (AEC1s) line airspaces and mediate gas exchange. After injury, they are regenerated by differentiation ... ...

Abstract	The lung is a relatively quiescent organ during homeostasis, but has a remarkable capacity for repair after injury. Alveolar epithelial type I cells (AEC1s) line airspaces and mediate gas exchange. After injury, they are regenerated by differentiation from their progenitors - alveolar epithelial type II cells (AEC2s) - which also secrete surfactant to maintain surface tension and alveolar patency. While recent studies showed that the maintenance of AEC2 stemness is Wnt dependent, the molecular mechanisms underlying AEC2-AEC1 differentiation in adult lung repair are still incompletely understood. Here we show that WWTR1 (TAZ) plays a crucial role in AEC differentiation. Using an in vitro organoid culture system, we found that tankyrase inhibition can efficiently block AEC2-AEC1 differentiation, and this effect was due to the inhibition of TAZ. In a bleomycin induced lung injury model, conditional deletion of TAZ in AEC2s dramatically reduced AEC1 regeneration during recovery, leading to exacerbated alveolar lesions and fibrosis. In patients with idiopathic pulmonary fibrosis (IPF), decreased blood levels of RAGE, a biomarker of AEC1 health, were associated with more rapid disease progression. Our findings implicate TAZ as a critical factor involved in AEC2 to AEC1 differentiation, and hence the maintenance of alveolar integrity after injury.
Mesh-Begriff(e)	Alveolar Epithelial Cells/drug effects ; Alveolar Epithelial Cells/metabolism ; Animals ; Bleomycin/adverse effects ; Cell Differentiation/drug effects ; Cell Differentiation/physiology ; Disease Models, Animal ; Humans ; Idiopathic Pulmonary Fibrosis/pathology ; Lung/pathology ; Lung Injury/chemically induced ; Lung Injury/metabolism ; Lung Injury/pathology ; Mice, Inbred C57BL ; Mice, Knockout ; Organogenesis/drug effects ; Organogenesis/physiology ; Organoids/metabolism ; Regeneration/physiology ; Stem Cells/metabolism ; Trans-Activators/genetics ; Trans-Activators/metabolism ; Trans-Activators/pharmacology ; Transcriptome ; beta Catenin/genetics ; beta Catenin/metabolism
Chemische Substanzen	Trans-Activators ; Wwtr1 protein, mouse ; beta Catenin ; Bleomycin (11056-06-7)
Sprache	Englisch
Erscheinungsdatum	2019-06-18
Erscheinungsland	United States
Dokumenttyp	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.128674
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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