Two-photon fluorescent turn-on probes for highly efficient detection and profiling of thiols in live cells and tissues

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Congzhen Shen ◽  
Duoteng Zhang ◽  
Fang Xu ◽  
Yang Yang ◽  
Yi Tan ◽  
...  
Keyword(s):  
Live Cells ◽  
Dual Purpose ◽  
Two Photon ◽  
Simultaneous Imaging ◽  
Turn On ◽  
Efficient Detection ◽  
Tumor Tissues ◽  
Cancer Diagnosis And Therapy

Abstract Thiols are important units in amino acids such as cysteine and peptides like glutathione. Development of chemical sensors capable of precise detection of thiols is important in cancer diagnosis and therapy. We have developed novel two-photon fluorescent turn-on probes for selective detection of thiols. The probes displayed excellent sensitivity and low detection limits. The dual-purpose probes have been demonstrated to be suitable for simultaneous imaging and proteome profiling in live cells and tumor tissues. The unique turn-on design endows the probes with excellent selectivity toward thiols in vitro and in situ, and can be further developed to support a thiol-quantification assay.

Real-time in situ monitoring via europium emission of the photo-release of antitumor cisplatin from a Eu–Pt complex

2015 ◽  
Vol 51 (74) ◽  
pp. 14022-14025 ◽  
Author(s):  
Hongguang Li ◽  
Rongfeng Lan ◽  
Chi-Fai Chan ◽  
Lijun Jiang ◽  
Lixiong Dai ◽  
...  
Keyword(s):  
Real Time ◽  
Antitumor Agent ◽  
In Situ Monitoring ◽  
Two Photon ◽  
Turn On ◽  
Pt Complex ◽  
Photon Excitation ◽  
Two Photon Excitation

A light-responsive antitumor agent, PtEuL, has been synthesized and evaluated for controlled cisplatin release by linear/two-photon excitation in vitro with concomitant turn-on Eu emission as a responsive traceable signal.

Turn-on fluorescent probe for dopamine detection in solutions and live cells based on in situ formation of aminosilane-functionalized carbon dots

2021 ◽  
Vol 1157 ◽  
pp. 338394
Author(s):  
Xiao-Yue Tang ◽  
Yi-Ming Liu ◽  
Xiao-Lin Bai ◽  
Hao Yuan ◽  
Yi-Kao Hu ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Carbon Dots ◽  
Live Cells ◽  
Turn On ◽  
Dopamine Detection ◽  
In Situ Formation

Ligand-displacement-based two-photon fluorogenic probe for visualizing mercapto biomolecules in live cells, Drosophila brains and zebrafish

The Analyst ◽  
2018 ◽  
Vol 143 (14) ◽  
pp. 3433-3441 ◽  
Author(s):  
Yanfei Zhao ◽  
Yun Ni ◽  
Liulin Wang ◽  
Chenchen Xu ◽  
Chenqi Xin ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Live Cell ◽  
Live Cells ◽  
Two Photon ◽  
Ligand Displacement ◽  
Cell Tissue ◽  
Fluorogenic Probe ◽  
Displacement Based

We report the Fe(iii)-based complex TPFeS which acts as a novel ligand-displacement-based TP fluorogenic probe for the rapid detection of mercapto biomolecules both in vitro and in live cell/tissue/in vivo imaging.

scholarly journals Measurement and analysis of sarcomere length in rat cardiomyocytes in situ and in vitro

2010 ◽  
Vol 298 (5) ◽  
pp. H1616-H1625 ◽  
Author(s):  
G. Bub ◽  
P. Camelliti ◽  
C. Bollensdorff ◽  
D. J. Stuckey ◽  
G. Picton ◽  
...  
Keyword(s):  
Measurement Errors ◽  
Sarcomere Length ◽  
Ex Vivo ◽  
Living Cells ◽  
Perfused Heart ◽  
Experimental Conditions ◽  
Rat Cardiomyocytes ◽  
Two Photon

Sarcomere length (SL) is an important determinant and indicator of cardiac mechanical function; however, techniques for measuring SL in living, intact tissue are limited. Here, we present a technique that uses two-photon microscopy to directly image striations of living cells in cardioplegic conditions, both in situ (Langendorff-perfused rat hearts and ventricular tissue slices, stained with the fluorescent marker di-4-ANEPPS) and in vitro (acutely isolated rat ventricular myocytes). Software was developed to extract SL from two-photon fluorescence image sets while accounting for measurement errors associated with motion artifact in raster-scanned images and uncertainty of the cell angle relative to the imaging plane. Monte-Carlo simulations were used to guide analysis of SL measurements by determining error bounds as a function of measurement path length. The mode of the distribution of SL measurements in resting Langendorff-perfused heart is 1.95 μm ( n = 167 measurements from N = 11 hearts) after correction for tissue orientation, which was significantly greater than that in isolated cells (1.71 μm, n = 346, N = 9 isolations) or ventricular slice preparations (1.79 μm, n = 79, N = 3 hearts) under our experimental conditions. Furthermore, we find that edema in arrested Langendorff-perfused heart is associated with a mean SL increase; this occurs as a function of time ex vivo and correlates with tissue volume changes determined by magnetic resonance imaging. Our results highlight that the proposed method can be used to monitor SL in living cells and that different experimental models from the same species may display significantly different SL values under otherwise comparable conditions, which has implications for experiment design, as well as comparison and interpretation of data.

Two-photon background-free fluorescence assay for glutathione over cysteine and homocysteine in vitro and vivo

2020 ◽  
Vol 56 (47) ◽  
pp. 6380-6383
Author(s):  
Qian Zhang ◽  
Yuzhe Xiao ◽  
Manqing Su ◽  
Peng Zhang ◽  
Yanling Gong ◽  
...  
Keyword(s):  
Fluorescence Assay ◽  
Sensory Receptor ◽  
Two Photon

A background-free fluorescent sensory receptor, with potential to undergo an in situ sensing strategy with new chromophores generated upon the detection event, was designed for the detection of glutathione.

scholarly journals Chemical profiling of DNA G-quadruplex-interacting proteins in live cells

2021 ◽  
Author(s):  
Xiaoyun Zhang ◽  
Jochen Spiegel ◽  
Sergio Martínez Cuesta ◽  
Santosh Adhikari ◽  
Shankar Balasubramanian
Keyword(s):  
Protein Interactions ◽  
Protein Profiling ◽  
Live Cells ◽  
Interacting Proteins ◽  
Chemical Profiling ◽  
G Quadruplex ◽  
Functional Classes ◽  
Chemical Strategy

AbstractDNA–protein interactions regulate critical biological processes. Identifying proteins that bind to specific, functional genomic loci is essential to understand the underlying regulatory mechanisms on a molecular level. Here we describe a co-binding-mediated protein profiling (CMPP) strategy to investigate the interactome of DNA G-quadruplexes (G4s) in native chromatin. CMPP involves cell-permeable, functionalized G4-ligand probes that bind endogenous G4s and subsequently crosslink to co-binding G4-interacting proteins in situ. We first showed the robustness of CMPP by proximity labelling of a G4 binding protein in vitro. Employing this approach in live cells, we then identified hundreds of putative G4-interacting proteins from various functional classes. Next, we confirmed a high G4-binding affinity and selectivity for several newly discovered G4 interactors in vitro, and we validated direct G4 interactions for a functionally important candidate in cellular chromatin using an independent approach. Our studies provide a chemical strategy to map protein interactions of specific nucleic acid features in living cells.

Specific receptor for hydrazine: mapping the in situ release of hydrazine in live cells and in an in vitro enzymatic assay

2016 ◽  
Vol 52 (36) ◽  
pp. 6166-6169 ◽  
Author(s):  
Firoj Ali ◽  
Anila H. A. ◽  
Nandaraj Taye ◽  
Devraj G. Mogare ◽  
Samit Chattopadhyay ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Enzymatic Assay ◽  
Live Cells ◽  
Specific Receptor ◽  
Specific Detection ◽  
Physiological Conditions

New chemodosimetric reagent for the specific detection of hydrazine in physiological conditions as well as for the mapping of its in situ generation in live Hct116 and HepG2 cells by enzymatic transformations.

scholarly journals Ligand affinity of the 67-kD elastin/laminin binding protein is modulated by the protein's lectin domain: visualization of elastin/laminin-receptor complexes with gold-tagged ligands.

1991 ◽  
Vol 113 (1) ◽  
pp. 187-194 ◽  
Author(s):  
R P Mecham ◽  
L Whitehouse ◽  
M Hay ◽  
A Hinek ◽  
M P Sheetz
Keyword(s):  
Cell Surface ◽  
Laminin Receptor ◽  
Live Cells ◽  
Receptor Interaction ◽  
Ligand Complex ◽  
Lectin Domain ◽  
Receptor Complexes ◽  
Laminin Binding Protein

Video-enhanced microscopy was used to examine the interaction of elastin- or laminin-coated gold particles with elastin binding proteins on the surface of live cells. By visualizing the binding events in real time, it was possible to determine the specificity and avidity of ligand binding as well as to analyze the motion of the receptor-ligand complex in the plane of the plasma membrane. Although it was difficult to interpret the rates of binding and release rigorously because of the possibility for multiple interactions between particles and the cell surface, relative changes in binding have revealed important aspects of the regulation of affinity of ligand-receptor interaction in situ. Both elastin and laminin were found to compete for binding to the cell surface and lactose dramatically decreased the affinity of the receptor(s) for both elastin and laminin. These findings were supported by in vitro studies of the detergent-solubilized receptor. Further, immobilization of the ligand-receptor complexes through binding to the cytoskeleton dramatically decreased the ability of bound particles to leave the receptor. The changes in the kinetics of ligand-coated gold binding to living cells suggest that both laminin and elastin binding is inhibited by lactose and that attachment of receptor to the cytoskeleton increases its affinity for the ligand.

scholarly journals Tracking RNA with light: selection, structure, and design of fluorescence turn-on RNA aptamers

2019 ◽  
Vol 52 ◽  
Author(s):  
Robert J. Trachman ◽  
Adrian R. Ferré-D'Amaré
Keyword(s):  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Structural Diversity ◽  
Rna Aptamers ◽  
Live Cells ◽  
Rna Molecules ◽  
Turn On ◽  
Green Fluorescent ◽  
Fluorescence Turn On

AbstractFluorescence turn-on aptamers,in vitroevolved RNA molecules that bind conditional fluorophores and activate their fluorescence, have emerged as RNA counterparts of the fluorescent proteins. Turn-on aptamers have been selected to bind diverse fluorophores, and they achieve varying degrees of specificity and affinity. These RNA–fluorophore complexes, many of which exceed the brightness of green fluorescent protein and their variants, can be used as tags for visualizing RNA localization and transport in live cells. Structure determination of several fluorescent RNAs revealed that they have diverse, unrelated overall architectures. As most of these RNAs activate the fluorescence of their ligands by restraining their photoexcited states into a planar conformation, their fluorophore binding sites have in common a planar arrangement of several nucleobases, most commonly a G-quartet. Nonetheless, each turn-on aptamer has developed idiosyncratic structural solutions to achieve specificity and efficient fluorescence turn-on. The combined structural diversity of fluorophores and turn-on RNA aptamers has already produced combinations that cover the visual spectrum. Further molecular evolution and structure-guided engineering is likely to produce fluorescent tags custom-tailored to specific applications.

scholarly journals Cancer-Specific hNQO1-Responsive Biocompatible Naphthalimides Providing a Rapid Fluorescent Turn-On with an Enhanced Enzyme Affinity

Sensors ◽  
2019 ◽  
Vol 20 (1) ◽  
pp. 53 ◽  
Author(s):  
Sun Young Park ◽  
Eugeine Jung ◽  
Jong Seung Kim ◽  
Sung-Gil Chi ◽  
Min Hee Lee
Keyword(s):  
Cancer Cells ◽  
Fluorescent Probes ◽  
Therapeutic Agent ◽  
Cell Imaging ◽  
Physiological Condition ◽  
Live Cells ◽  
Strong Fluorescence ◽  
Turn On ◽  
Efficient Detection ◽  
Agent Development

Human NAD(P)H:quinone oxidoreductase 1 (hNQO1) is overexpressed in cancer cells and associated with the drug resistance factor of cancer. The objective of this work is the development of fluorescent probes for the efficient detection of hNQO1 activity in cancer cells, which can be employed for the cancer diagnosis and therapeutic agent development. Herein, we report naphthalimide-based fluorescent probes 1 and 2 that can detect hNQO1. For hNQO1 activity, the probes showed a significant fluorescence increase at 540 nm. In addition, probe 1, the naphthalimide containing a triphenylphosphonium salt, showed an enhanced enzyme efficiency and rapid detection under a physiological condition. The detection ability of probe 1 was superior to that of other previously reported probes. Moreover, probe 1 was less cytotoxic during the cancer cell imaging and readily provided a strong fluorescence in hNQO1-overexpressed cancer cells (A549). We proposed that probe 1 can be used to detect hNQO1 expression in live cells and it will be applied to develop the diagnosis and customized treatment of hNQO1-related disease.

Sign in / Sign up

Export Citation Format

Share Document