scholarly journals Simultaneous recording of multiple cellular signaling events by frequency- and spectrally-tuned multiplexing of fluorescent probes

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Michelina Kierzek ◽  
Parker E Deal ◽  
Evan W Miller ◽  
Shatanik Mukherjee ◽  
Dagmar Wachten ◽  
...  
Keyword(s):  
Time Resolution ◽  
Fluorescent Probes ◽  
Cellular Signaling ◽  
High Sensitivity ◽  
Simultaneous Recording ◽  
Rapid Changes ◽  
Fret Biosensors ◽  
Phase Sensitive ◽  
Small Biomolecules ◽  
Cell Microscopy

Fluorescent probes that change their spectral properties upon binding to small biomolecules, ions, or changes in the membrane potential (Vm) are invaluable tools to study cellular signaling pathways. Here, we introduce a novel technique for simultaneous recording of multiple probes at millisecond time resolution: frequency- and spectrally-tuned multiplexing (FASTM). Different from present multiplexing approaches, FASTM uses phase-sensitive signal detection, which renders various combinations of common probes for Vm and ions accessible for multiplexing. Using kinetic stopped-flow fluorimetry, we show that FASTM allows simultaneous recording of rapid changes in Ca2+, pH, Na+, and Vm with high sensitivity and minimal crosstalk. FASTM is also suited for multiplexing using single-cell microscopy and genetically-encoded FRET biosensors. Moreover, FASTM is compatible with opto-chemical tools to study signaling using light. Finally, we show that the exceptional time resolution of FASTM also allows resolving rapid chemical reactions. Altogether, FASTM opens new opportunities for interrogating cellular signaling.

scholarly journals Fluorescent Probes for Live Cell Thiol Detection

Molecules ◽  
2021 ◽  
Vol 26 (12) ◽  
pp. 3575
Author(s):  
Shenggang Wang ◽  
Yue Huang ◽  
Xiangming Guan
Keyword(s):  
Fluorescent Probes ◽  
Biological System ◽  
High Sensitivity ◽  
Intracellular Distribution ◽  
Live Cell ◽  
Live Cells ◽  
High Demand ◽  
Non Invasive

Thiols play vital and irreplaceable roles in the biological system. Abnormality of thiol levels has been linked with various diseases and biological disorders. Thiols are known to distribute unevenly and change dynamically in the biological system. Methods that can determine thiols’ concentration and distribution in live cells are in high demand. In the last two decades, fluorescent probes have emerged as a powerful tool for achieving that goal for the simplicity, high sensitivity, and capability of visualizing the analytes in live cells in a non-invasive way. They also enable the determination of intracellular distribution and dynamitic movement of thiols in the intact native environments. This review focuses on some of the major strategies/mechanisms being used for detecting GSH, Cys/Hcy, and other thiols in live cells via fluorescent probes, and how they are applied at the cellular and subcellular levels. The sensing mechanisms (for GSH and Cys/Hcy) and bio-applications of the probes are illustrated followed by a summary of probes for selectively detecting cellular and subcellular thiols.

scholarly journals Monitoring of a Highly Flexible Aircraft Model Wing Using Time-Expanded Phase-Sensitive OTDR

Sensors ◽  
2021 ◽  
Vol 21 (11) ◽  
pp. 3766
Author(s):  
Miguel Soriano-Amat ◽  
David Fragas-Sánchez ◽  
Hugo F. Martins ◽  
David Vallespín-Fontcuberta ◽  
Javier Preciado-Garbayo ◽  
...  
Keyword(s):  
Optical Fibers ◽  
Fuel Efficiency ◽  
Optical Fiber Sensor ◽  
High Sensitivity ◽  
Time Domain Reflectometry ◽  
Structural Deformation ◽  
Flexible Wings ◽  
Sensing Technology ◽  
Distributed Optical Fiber Sensor ◽  
Phase Sensitive

In recent years, the use of highly flexible wings in aerial vehicles (e.g., aircraft or drones) has been attracting increasing interest, as they are lightweight, which can improve fuel-efficiency and distinct flight performances. Continuous wing monitoring can provide valuable information to prevent fatal failures and optimize aircraft control. In this paper, we demonstrate the capabilities of a distributed optical fiber sensor based on time-expanded phase-sensitive optical time-domain reflectometry (TE-ΦOTDR) technology for structural health monitoring of highly flexible wings, including static (i.e., bend and torsion), and dynamic (e.g., vibration) structural deformation. This distributed sensing technology provides a remarkable spatial resolution of 2 cm, with detection and processing bandwidths well under the MHz, arising as a novel, highly efficient monitoring methodology for this kind of structure. Conventional optical fibers were embedded in two highly flexible specimens that represented an aircraft wing, and different bending and twisting movements were detected and quantified with high sensitivity and minimal intrusiveness.

scholarly journals Biological nanoscale fluorescent probes: From structure and performance to bioimaging

2020 ◽  
Vol 39 (1) ◽  
pp. 209-221
Author(s):  
Jiafeng Wan ◽  
Xiaoyuan Zhang ◽  
Kai Zhang ◽  
Zhiqiang Su
Keyword(s):  
Fluorescent Probes ◽  
Organic Dyes ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Biological Imaging ◽  
Fluorescent Materials ◽  
And Performance

Abstract In recent years, nanomaterials have attracted lots of attention from researchers due to their unique properties. Nanometer fluorescent materials, such as organic dyes, semiconductor quantum dots (QDs), metal nano-clusters (MNCs), carbon dots (CDs), etc., are widely used in biological imaging due to their high sensitivity, short response time, and excellent accuracy. Nanometer fluorescent probes can not only perform in vitro imaging of organisms but also achieve in vivo imaging. This provides medical staff with great convenience in cancer treatment. Combined with contemporary medical methods, faster and more effective treatment of cancer is achievable. This article explains the response mechanism of three-nanometer fluorescent probes: the principle of induced electron transfer (PET), the principle of fluorescence resonance energy transfer (FRET), and the principle of intramolecular charge transfer (ICT), showing the semiconductor QDs, precious MNCs, and CDs. The excellent performance of the three kinds of nano fluorescent materials in biological imaging is highlighted, and the application of these three kinds of nano fluorescent probes in targeted biological imaging is also introduced. Nanometer fluorescent materials will show their significance in the field of biomedicine.

scholarly journals Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching

PLoS ONE ◽  
2016 ◽  
Vol 11 (1) ◽  
pp. e0147115 ◽  
Author(s):  
Rumelo Amor ◽  
Alison McDonald ◽  
Johanna Trägårdh ◽  
Gillian Robb ◽  
Louise Wilson ◽  
...  
Keyword(s):  
Time Resolution ◽  
Live Cell ◽  
High Time ◽  
High Time Resolution ◽  
Two Photon ◽  
Photon Excitation ◽  
Live Cell Microscopy ◽  
Cell Microscopy ◽  
Two Photon Excitation

scholarly journals Green preparation of nitrogen doped carbon quantum dot films as fluorescent probes

RSC Advances ◽  
2017 ◽  
Vol 7 (88) ◽  
pp. 56087-56092 ◽  
Author(s):  
Yanfei He ◽  
Lina Liang ◽  
Qinghao Liu ◽  
Jinchun Guo ◽  
Dong Liang ◽  
...  
Keyword(s):  
Carbon Source ◽  
Quantum Dot ◽  
Nitrogen Source ◽  
Fluorescence Intensity ◽  
Fluorescent Probes ◽  
High Sensitivity ◽  
Nitrogen Doped ◽  
Carbon Quantum Dot ◽  
Sensitivity And Selectivity ◽  
Nitrogen Doped Carbon

A facile and economical hydrothermal method was developed for the preparation of highly luminescent NCDs by using cabbage juice as carbon source and PP as nitrogen source. The fluorescence intensity of CA-NCDs was quenched by Fe3+ with high sensitivity and selectivity.

scholarly journals Ultraviolet Carbon Nanodots Providing a Dual-Mode Spectral Matching Platform for Synergistic Enhancement of the Fluorescent Sensing

Molecules ◽  
2020 ◽  
Vol 25 (11) ◽  
pp. 2679
Author(s):  
Liman Sai ◽  
Shuping Jiao ◽  
Jianwen Yang
Keyword(s):  
Fluorescent Probes ◽  
High Sensitivity ◽  
Inner Filter Effect ◽  
Carbon Nanodots ◽  
Spectral Matching ◽  
Fluorescent Sensing ◽  
Chromium Vi ◽  
Band Emission ◽  
Synergistic Enhancement ◽  
Emissive Property

The sensing of chromium(VI) (Cr(VI)) is highly desired, due to its toxic and carcinogenic effects upon human health. Fluorescent probes, especially carbon nanodots (CNDs), have been widely used for Cr(VI) sensing via the inner filter effect (IFE). However, improving the sensitivity of these probes remains a difficult issue. In this work, CNDs derived from β-Lactoglobulin were applied as an ultrasensitive fluorescent probe for Cr(VI). With 260 nm excitation, the CNDs showed multi-band emission, including an ultraviolet 360 nm peak. The spectral matching of the CNDs with Cr(VI) led to synergistic suppression of both the excitation and emission light in the fluorescent sensing. As a consequence, the CNDs showed high sensitivity toward Cr(VI), the detection limit reaching as low as 20 nM. Moreover, taking advantage of the multi-emissive property of the CNDs, the synergistic effect was proven in an IFE-based sensing system, which might be extended to the design of other kinds of fluorescent probes.

scholarly journals Far-red pentamethine cyanine dyes as fluorescent probes for the detection of serum albumins

2020 ◽  
Vol 7 (7) ◽  
pp. 200453
Author(s):  
D. Aristova ◽  
G. Volynets ◽  
S. Chernii ◽  
M. Losytskyy ◽  
A. Balanda ◽  
...  
Keyword(s):  
Binding Site ◽  
Fluorescent Probes ◽  
Protein Detection ◽  
High Sensitivity ◽  
Luminescent Properties ◽  
Cyanine Dyes ◽  
Globular Proteins ◽  
Sulfonate Group ◽  
Serum Albumins ◽  
Pentamethine Cyanine Dyes

Benzothiazole based cyanine dyes with bridged groups in the pentamethine chain were studied as potential far-red fluorescent probes for protein detection. Spectral-luminescent properties were characterized for unbound dyes and in the presence of serum albumins (bovine (BSA), human (HSA), equine (ESA)), and globular proteins (β-lactoglobulin, ovalbumin). We have observed that the addition of albumins leads to a significant increase in dyes fluorescence intensity. However, the fluorescent response of dyes in the presence of other globular proteins was notably lower. The value of fluorescence quantum yield for dye bearing a sulfonate group complexed with HSA amounted to 42% compared with 0.2% for the free dye. The detection limit of HSA by this dye was greater than 0.004 mg ml −1 which indicates the high sensitivity of dye to low HSA concentrations. Modelling of structure of the dyes complexes with albumin molecules was performed by molecular docking. According to these data, dyes could bind to up to five sites on the HSA molecule; the most preferable are the haemin-binding site in subdomain IB and the dye-binding site in the pocket between subdomains IA, IIA and IIIA. This work confirms that pentamethine cyanine dyes could be proposed as powerful far-red fluorescent probes applicable for highly sensitive detection of albumins.

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