Ratiometric fluorescence imaging of endogenous selenocysteine in cancer cell matrix

2017 ◽  
Vol 5 (33) ◽  
pp. 6890-6896 ◽  
Author(s):  
Yong Tian ◽  
Fangyun Xin ◽  
Congcong Gao ◽  
Jing Jing ◽  
Xiaoling Zhang
Keyword(s):  
Fluorescence Imaging ◽  
Cancer Cell ◽  
Blue Emission ◽  
Yellow Emission ◽  
Fluorescence Ratio ◽  
Ratiometric Fluorescence ◽  
Specific Reaction ◽  
Quantitative Correlation ◽  
Cell Matrix ◽  
A Cell

A ratiometric method for tracing the variation of selenocysteine (Sec) in a cell matrix was provided. This was constructed by the quantitative correlation between the fluorescence ratio ofRat-Sec(blue emission, the probe) andNap-OH(green-yellow emission, the product from a Sec-specific reaction) and the concentration of Sec.

A DNA-functionalized biomass nanoprobe for the targeted photodynamic therapy of tumor and ratiometric fluorescence imaging-based visual cancer cell identification/antitumor drug screening

The Analyst ◽  
2021 ◽  
Author(s):  
Pengxiang Lin ◽  
Liangliang Zhang ◽  
Dongxia Chen ◽  
Jiayao Xu ◽  
Yulong Bai ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Fluorescence Imaging ◽  
Cancer Cell ◽  
Drug Screening ◽  
Antitumor Drug ◽  
Ratiometric Fluorescence ◽  
Cell Identification ◽  
Targeted Photodynamic Therapy

A DNA-functionalized biomass nanoprobe was developed for the targeted photodynamic therapy of tumor and ratiometric fluorescence imaging-based visual cancer cell identification/antitumor drug screening.

scholarly journals A Dual-Fluorophore Sensor Approach for Ratiometric Fluorescence Imaging of Potassium in Living Cells

2020 ◽  
Author(s):  
Zeming Wang ◽  
Tyler C. Detomasi ◽  
Christopher Chang
Keyword(s):  
Fluorescence Imaging ◽  
Cell Lines ◽  
Cancer Cell ◽  
High Selectivity ◽  
Cancer Cell Lines ◽  
Biological Evaluation ◽  
The Body ◽  
Ratiometric Fluorescence ◽  
Intracellular Potassium ◽  
Ratiometric Detection

Potassium is the most abundant intracellular metal in the body, playing vital roles in regulating intracellular fluid volume, nutrient transport, and cell-to-cell communication through nerve and muscle contraction. On the other hand, aberrant alterations in K<sup>+</sup> homeostasis contribute to a diverse array of diseases spanning cardiovascular and neurological disorders to diabetes to kidney disease to cancer. Owing to the large differences in intracellular versus extracellular K<sup>+</sup> concentrations ([K<sup>+</sup>]<sub>intra</sub> = 150 mM, [K<sup>+</sup>]<sub>extra</sub> = 3-5 mM), an unmet need for studies of K<sup>+</sup> physiology and pathology remains a relative dearth of methods to reliably measure dynamic changes in intracellular K<sup>+</sup> in biological specimens that meet the dual challenges of low affinity and high selectivity for K<sup>+</sup>, particularly over Na<sup>+</sup>, as currently available fluorescent K<sup>+</sup> sensors are largely optimized with high-affinity receptors that are more amenable for extracellular K<sup>+</sup> detection. We report the design, synthesis, and biological evaluation of Ratiometric Potassium Sensor 1 (<b>RPS-1</b>), a dual-fluorophore sensor that enables ratiometric fluorescence imaging of intracellular potassium in living systems. <b>RPS-1</b> links a potassium-responsive fluorescent sensor fragment (<b>PS525</b>) with a low-affinity, high-selectivity crown ether receptor for K<sup>+</sup> to a potassium-insensitive reference fluorophore (<b>Coumarin 343</b>) as an internal calibration standard through ester bonds. Upon intracellular delivery, esterase-directed cleavage splits these two dyes into separate fragments to enable ratiometric detection of K<sup>+</sup>. <b>RPS-1</b> responds to K<sup>+</sup> in aqueous buffer with high selectivity over competing metal ions and is sensitive to potassium ions at steady-state intracellular levels and can respond to decreases or increases from that basal set point. Moreover, <b>RPS-1</b> was applied for comparative screening of K<sup>+</sup> pools across a panel of different cancer cell lines, revealing elevations in basal intracellular K<sup>+</sup> in metastatic breast cancer cell lines vs normal breast cells. This work provides a unique chemical tool for the study of intracellular potassium dynamics and a starting point for the design of other ratiometric fluorescent sensors based on two-fluorophore approaches that do not rely on FRET or related energy transfer designs.

scholarly journals A Dual-Fluorophore Sensor Approach for Ratiometric Fluorescence Imaging of Potassium in Living Cells

2020 ◽  
Author(s):  
Zeming Wang ◽  
Tyler C. Detomasi ◽  
Christopher Chang
Keyword(s):  
Fluorescence Imaging ◽  
Cell Lines ◽  
Cancer Cell ◽  
High Selectivity ◽  
Cancer Cell Lines ◽  
Biological Evaluation ◽  
The Body ◽  
Ratiometric Fluorescence ◽  
Intracellular Potassium ◽  
Ratiometric Detection

Potassium is the most abundant intracellular metal in the body, playing vital roles in regulating intracellular fluid volume, nutrient transport, and cell-to-cell communication through nerve and muscle contraction. On the other hand, aberrant alterations in K<sup>+</sup> homeostasis contribute to a diverse array of diseases spanning cardiovascular and neurological disorders to diabetes to kidney disease to cancer. Owing to the large differences in intracellular versus extracellular K<sup>+</sup> concentrations ([K<sup>+</sup>]<sub>intra</sub> = 150 mM, [K<sup>+</sup>]<sub>extra</sub> = 3-5 mM), an unmet need for studies of K<sup>+</sup> physiology and pathology remains a relative dearth of methods to reliably measure dynamic changes in intracellular K<sup>+</sup> in biological specimens that meet the dual challenges of low affinity and high selectivity for K<sup>+</sup>, particularly over Na<sup>+</sup>, as currently available fluorescent K<sup>+</sup> sensors are largely optimized with high-affinity receptors that are more amenable for extracellular K<sup>+</sup> detection. We report the design, synthesis, and biological evaluation of Ratiometric Potassium Sensor 1 (<b>RPS-1</b>), a dual-fluorophore sensor that enables ratiometric fluorescence imaging of intracellular potassium in living systems. <b>RPS-1</b> links a potassium-responsive fluorescent sensor fragment (<b>PS525</b>) with a low-affinity, high-selectivity crown ether receptor for K<sup>+</sup> to a potassium-insensitive reference fluorophore (<b>Coumarin 343</b>) as an internal calibration standard through ester bonds. Upon intracellular delivery, esterase-directed cleavage splits these two dyes into separate fragments to enable ratiometric detection of K<sup>+</sup>. <b>RPS-1</b> responds to K<sup>+</sup> in aqueous buffer with high selectivity over competing metal ions and is sensitive to potassium ions at steady-state intracellular levels and can respond to decreases or increases from that basal set point. Moreover, <b>RPS-1</b> was applied for comparative screening of K<sup>+</sup> pools across a panel of different cancer cell lines, revealing elevations in basal intracellular K<sup>+</sup> in metastatic breast cancer cell lines vs normal breast cells. This work provides a unique chemical tool for the study of intracellular potassium dynamics and a starting point for the design of other ratiometric fluorescent sensors based on two-fluorophore approaches that do not rely on FRET or related energy transfer designs.

Digital Technologies – Innovative Solutions for Agriculture

2020 ◽  
Vol 67 (1) ◽  
pp. 127-132
Author(s):  
Marina A. Nikitina ◽  
Dmitriy N. Osyanin ◽  
Irina V. Petrunina
Keyword(s):  
Farm Animals ◽  
High Tech ◽  
Mathematical Apparatus ◽  
Online Mode ◽  
Cell Matrix ◽  
Algorithmic Support ◽  
Quality Product ◽  
A Cell ◽  
Parametric Description ◽  
Animal Feeding

High-tech decision support systems based on the knowledge of experts in the field of animal feeding and a rigorous mathematical apparatus make it possible to increase the productivity of the animals and provide a high-quality product with the ability to trace the trophological chain from the field to the counter. (Research purpose) To identify methodological approaches for the development of algorithmic support and software for assessment and optimizing the animals diet. (Materials and Methods) The authors complied a structural-parametric model of a productive animal feeding in a cell matrix form. They represented the whole variety of existing known and unknown relationships between the factors of the farm animal state and the feeding ration characteristics. It was found that when forming a feeding ration, it was necessary to assess the degree of influence on productivity of not individual indicators, but its combination. (Results and discussion) The authors considered parametric descriptions of farm animals; diet with selected groups of indicators, characteristics and properties. It was stated that the parametric description of feeding contained a set of parameters taking into account at least 25 nutritional indicators. They presented a mathematical apparatus for evaluating and optimizing the feeding ration. It was determined that the automation of the process of calculating the daily ration of an animal allowed to change the diets depending on the availability of feed in the farm and their chemical composition in real time. The authors found out that an increase in the productivity of animals and poultry by 30-50 percent was possible due to balanced full-fledged feeding. (Conclusions) The authors showed a systematic approach to the development of software for evaluating and optimizing the animal feeding diet. It was revealed that a computer system in the online mode would help to find the best option for a feeding ration, taking into account multifactor and limitations.

scholarly journals Self-Assembling Polypeptide Hydrogels as a Platform to Recapitulate the Tumor Microenvironment

Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3286
Author(s):  
Dariusz Lachowski ◽  
Carlos Matellan ◽  
Ernesto Cortes ◽  
Alberto Saiani ◽  
Aline F. Miller ◽  
...  
Keyword(s):  
Cell Culture ◽  
Tumor Microenvironment ◽  
Cancer Cell ◽  
Critical Role ◽  
Hypoxia Inducible Factor ◽  
Pancreatic Cancer Cell Line ◽  
Cancer Cell Line ◽  
Culture Systems ◽  
A Cell

The tumor microenvironment plays a critical role in modulating cancer cell migration, metabolism, and malignancy, thus, highlighting the need to develop in vitro culture systems that can recapitulate its abnormal properties. While a variety of stiffness-tunable biomaterials, reviewed here, have been developed to mimic the rigidity of the tumor extracellular matrix, culture systems that can recapitulate the broader extracellular context of the tumor microenvironment (including pH and temperature) remain comparably unexplored, partially due to the difficulty in independently tuning these parameters. Here, we investigate a self-assembled polypeptide network hydrogel as a cell culture platform and demonstrate that the culture parameters, including the substrate stiffness, extracellular pH and temperature, can be independently controlled. We then use this biomaterial as a cell culture substrate to assess the effect of stiffness, pH and temperature on Suit2 cells, a pancreatic cancer cell line, and demonstrate that these microenvironmental factors can regulate two critical transcription factors in cancer: yes-associated protein 1 (YAP) and hypoxia inducible factor (HIF-1A).

scholarly journals Nanonet force microscopy for measuring forces in single smooth muscle cells of the human aorta

2017 ◽  
Vol 28 (14) ◽  
pp. 1894-1900 ◽  
Author(s):  
Alexander Hall ◽  
Patrick Chan ◽  
Kevin Sheets ◽  
Matthew Apperson ◽  
Christopher Delaughter ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
External Perturbation ◽  
Human Aorta ◽  
Cycle Times ◽  
Force Microscopy ◽  
Cell Matrix ◽  
Innovative Methods ◽  
A Cell ◽  
Inside Out

A number of innovative methods exist to measure cell–matrix adhesive forces, but they have yet to accurately describe and quantify the intricate interplay of a cell and its fibrous extracellular matrix (ECM). In cardiovascular pathologies, such as aortic aneurysm, new knowledge on the involvement of cell–matrix forces could lead to elucidation of disease mechanisms. To better understand this dynamics, we measured primary human aortic single smooth muscle cell (SMC) forces using nanonet force microscopy in both inside-out (I-O intrinsic contractility) and outside-in (O-I external perturbation) modes. For SMC populations, we measured the I-O and O-I forces to be 12.9 ± 1.0 and 57.9 ± 2.5 nN, respectively. Exposure of cells to oxidative stress conditions caused a force decrease of 57 and 48% in I-O and O-I modes, respectively, and an increase in migration rate by 2.5-fold. Finally, in O-I mode, we cyclically perturbed cells at constant strain of varying duration to simulate in vivo conditions of the cardiac cycle and found that I-O forces decrease with increasing duration and O-I forces decreased by half at shorter cycle times. Thus our findings highlight the need to study forces exerted and felt by cells simultaneously to comprehensively understand force modulation in cardiovascular disease.

scholarly journals Adhesive multiplicity in the interaction of embryonic fibroblasts and myoblasts with extracellular matrices.

1984 ◽  
Vol 99 (4) ◽  
pp. 1398-1404 ◽  
Author(s):  
C Decker ◽  
R Greggs ◽  
K Duggan ◽  
J Stubbs ◽  
A Horwitz
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Muscle ◽  
Cardiac Fibroblasts ◽  
Cell Types ◽  
Extracellular Matrices ◽  
Common Denominator ◽  
Cell Matrix ◽  
Skeletal Myoblasts ◽  
A Cell ◽  
Cell Matrix Adhesion

Neff et al. (1982, J. Cell Biol., 95:654-666) have described a monoclonal antibody, CSAT, directed against a cell surface antigen that participates in the adhesion of skeletal muscle to extracellular matrices. We used the same antibody to compare and parse the determinants of adhesion and morphology on myogenic and fibrogenic cells. We report here that the antigen is present on skeletal and cardiac muscle and on tendon, skeletal, dermal, and cardiac fibroblasts; however, its contribution to their morphology and adhesion is different. The antibody produces large alterations in the morphology and adhesion of skeletal myoblasts and tendon fibroblasts; in contrast, its effects on the cardiac fibroblasts are not readily detected. The effects of CSAT on the other cell types, i.e., dermal and skeletal fibroblasts, cardiac muscle, 5-bromodeoxyuridine-treated skeletal muscle, lie between these extremes. The effects of CSAT on the skeletal myoblasts depends on the calcium concentration in the growth medium and on the culture age. We interpret these differential responses to CSAT as revealing differences in the adhesion of the various cells to extracellular matrices. This interpretation is supported by parallel studies using quantitative assays of cell-matrix adhesion. The likely origin of these adhesive differences is the progressive display of different kinds of adhesion-related molecules and their organizational complexes on increasingly adhesive cells. The antigen to which CSAT is directed is present on all of the above cells and thus appears to be a lowest common denominator of their adhesion to extracellular matrices.

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