scholarly journals Covalent and non-covalent coupling of a Au102 nanocluster with a fluorophore: energy transfer, quenching and intracellular pH sensing

2021 ◽  
Author(s):  
Eero Hulkko ◽  
Tanja Lahtinen ◽  
Varpu Marjomäki ◽  
Emmi Pohjolainen ◽  
Ville Saarnio ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Intracellular Ph ◽  
Fluorescent Dye ◽  
Live Cells ◽  
Ph Sensing ◽  
Gold Nanocluster ◽  
Covalent Coupling ◽  
Covalently Bound

A covalently bound hybrid between an atomically precise gold nanocluster and a fluorescent dye is loaded into cells via endocytosis. Acidification of the endosomes activates fluorescence, enabling optical pH sensing inside live cells.

Synthesis of gold nanocluster-loaded lysozyme nanoparticles for label-free ratiometric fluorescent pH sensing: applications to enzyme–substrate systems and cellular imaging

2019 ◽  
Vol 7 (24) ◽  
pp. 3876-3883 ◽  
Author(s):  
Shanmugam Chandirasekar ◽  
Jyun-Guo You ◽  
Jhe-Hong Xue ◽  
Wei-Lung Tseng
Keyword(s):  
Cellular Imaging ◽  
Live Cells ◽  
Ph Sensing ◽  
Label Free ◽  
Gold Nanocluster ◽  
Dual Emission ◽  
Ph Changes ◽  
Ratiometric Sensing ◽  
Sensing Applications ◽  
Enzyme Substrate

We have demonstrated the synthesis of gold nanocluster-loaded lysozyme nanoparticles as a dual-emission probe for ratiometric sensing of pH changes in enzyme–substrate systems and live cells.

A Small Molecule – Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence

2020 ◽  
Author(s):  
Brittany Benlian ◽  
Pavel Klier ◽  
Kayli Martinez ◽  
Marie Schwinn ◽  
Thomas Kirkland ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Membrane Potential ◽  
Small Molecule ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Potential Oscillations ◽  
Excitation Light ◽  
Voltage Imaging ◽  
Induced Pluripotent

<p>We report a small molecule enzyme pair for optical voltage sensing via quenching of bioluminescence. This <u>Q</u>uenching <u>B</u>ioluminescent V<u>olt</u>age Indicator, or Q-BOLT, pairs the dark absorbing, voltage-sensitive dipicrylamine with membrane-localized bioluminescence from the luciferase NanoLuc (NLuc). As a result, bioluminescence is quenched through resonance energy transfer (QRET) as a function of membrane potential. Fusion of HaloTag to NLuc creates a two-acceptor bioluminescence resonance energy transfer (BRET) system when a tetramethylrhodamine (TMR) HaloTag ligand is ligated to HaloTag. In this mode, Q-BOLT is capable of providing direct visualization of changes in membrane potential in live cells via three distinct readouts: change in QRET, BRET, and the ratio between bioluminescence emission and BRET. Q-BOLT can provide up to a 29% change in bioluminescence (ΔBL/BL) and >100% ΔBRET/BRET per 100 mV change in HEK 293T cells, without the need for excitation light. In cardiac monolayers derived from human induced pluripotent stem cells (hiPSC), Q-BOLT readily reports on membrane potential oscillations. Q-BOLT is the first example of a hybrid small molecule – protein voltage indicator that does not require excitation light and may be useful in contexts where excitation light is limiting.</p> <p> </p>

A Small Molecule – Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence

2020 ◽  
Author(s):  
Brittany Benlian ◽  
Pavel Klier ◽  
Kayli Martinez ◽  
Marie Schwinn ◽  
Thomas Kirkland ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Membrane Potential ◽  
Small Molecule ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Potential Oscillations ◽  
Excitation Light ◽  
Voltage Imaging ◽  
Induced Pluripotent

<p>We report a small molecule enzyme pair for optical voltage sensing via quenching of bioluminescence. This <u>Q</u>uenching <u>B</u>ioluminescent V<u>olt</u>age Indicator, or Q-BOLT, pairs the dark absorbing, voltage-sensitive dipicrylamine with membrane-localized bioluminescence from the luciferase NanoLuc (NLuc). As a result, bioluminescence is quenched through resonance energy transfer (QRET) as a function of membrane potential. Fusion of HaloTag to NLuc creates a two-acceptor bioluminescence resonance energy transfer (BRET) system when a tetramethylrhodamine (TMR) HaloTag ligand is ligated to HaloTag. In this mode, Q-BOLT is capable of providing direct visualization of changes in membrane potential in live cells via three distinct readouts: change in QRET, BRET, and the ratio between bioluminescence emission and BRET. Q-BOLT can provide up to a 29% change in bioluminescence (ΔBL/BL) and >100% ΔBRET/BRET per 100 mV change in HEK 293T cells, without the need for excitation light. In cardiac monolayers derived from human induced pluripotent stem cells (hiPSC), Q-BOLT readily reports on membrane potential oscillations. Q-BOLT is the first example of a hybrid small molecule – protein voltage indicator that does not require excitation light and may be useful in contexts where excitation light is limiting.</p> <p> </p>

scholarly journals Fluorescent Protein-Based Autophagy Biosensors

Materials ◽  
2021 ◽  
Vol 14 (11) ◽  
pp. 3019
Author(s):  
Heejung Kim ◽  
Jihye Seong
Keyword(s):  
Energy Transfer ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Treatment Strategies ◽  
Resonance Energy ◽  
Live Cells ◽  
Spectral Profile ◽  
Spatiotemporal Resolution ◽  
Future Directions ◽  
Complex Process

Autophagy is an essential cellular process of self-degradation for dysfunctional or unnecessary cytosolic constituents and organelles. Dysregulation of autophagy is thus involved in various diseases such as neurodegenerative diseases. To investigate the complex process of autophagy, various biochemical, chemical assays, and imaging methods have been developed. Here we introduce various methods to study autophagy, in particular focusing on the review of designs, principles, and limitations of the fluorescent protein (FP)-based autophagy biosensors. Different physicochemical properties of FPs, such as pH-sensitivity, stability, brightness, spectral profile, and fluorescence resonance energy transfer (FRET), are considered to design autophagy biosensors. These FP-based biosensors allow for sensitive detection and real-time monitoring of autophagy progression in live cells with high spatiotemporal resolution. We also discuss future directions utilizing an optobiochemical strategy to investigate the in-depth mechanisms of autophagy. These cutting-edge technologies will further help us to develop the treatment strategies of autophagy-related diseases.

BSA-Coated Nanoparticles for Improved SERS-Based Intracellular pH Sensing

2014 ◽  
Vol 86 (24) ◽  
pp. 12250-12257 ◽  
Author(s):  
Xiao-Shan Zheng ◽  
Pei Hu ◽  
Yan Cui ◽  
Cheng Zong ◽  
Jia-Min Feng ◽  
...  
Keyword(s):  
Intracellular Ph ◽  
Ph Sensing ◽  
Coated Nanoparticles

scholarly journals AIE-Driven Fluorescent Polysaccharide Polymersome as Enzyme-responsive FRET Nanoprobe to Study the Real-time Delivery Aspects in Live-Cells

2021 ◽  
Author(s):  
Nilesh Umakant Deshpande ◽  
Mishika Virmani ◽  
Manickam Jayakannan
Keyword(s):  
Energy Transfer ◽  
Confocal Microscopy ◽  
Real Time ◽  
Fluorescence Resonance Energy Transfer ◽  
Imaging Techniques ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Intracellular Enzyme ◽  
Bio Imaging

We report aggregation induced emission (AIE) driven polysaccharide polymersome as fluorescence resonance energy transfer (FRET) nanoprobes to study their intracellular enzyme-responsive delivery by real-time live-cell confocal microscopy bio-imaging techniques. AIE...

Reversible Self-Assembly of Nanoprobes in Live Cells for Dynamic Intracellular pH Imaging

ACS Nano ◽  
2019 ◽  
Author(s):  
Bo Dong ◽  
Shuangli Du ◽  
Chunxiao Wang ◽  
Haohao Fu ◽  
Qiang Li ◽  
...  
Keyword(s):  
Intracellular Ph ◽  
Self Assembly ◽  
Live Cells ◽  
Ph Imaging

Fluorescence-SERS dual-signal probes for pH sensing in live cells

2019 ◽  
Vol 562 ◽  
pp. 289-295 ◽  
Author(s):  
Guohai Yang ◽  
Qian Zhang ◽  
Yuan Liang ◽  
Hong Liu ◽  
Lu-Lu Qu ◽  
...  
Keyword(s):  
Live Cells ◽  
Ph Sensing
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