Fluorophore tagged bio-molecules and their applications: A brief review
TL;DR: This review demonstrates the applications of such conjugated fluorescent molecular probes in different domains of biological activities and brings out the advantages and disadvantages of this particular type of fluorophores with the insight to the future perspectives.
Abstract: Bio-molecules are principal building blocks of living species and their inter-play is the cause of bio-activities. Various sophisticated experimental techniques as well as theoretical studies have ...
Citations
More filters
01 Jan 2016
TL;DR: The topics in fluorescence spectroscopy is universally compatible with any devices to read, and is available in the digital library an online access to it is set as public so you can download it instantly.
Abstract: Thank you very much for reading topics in fluorescence spectroscopy. As you may know, people have search numerous times for their chosen books like this topics in fluorescence spectroscopy, but end up in infectious downloads. Rather than enjoying a good book with a cup of coffee in the afternoon, instead they are facing with some malicious virus inside their computer. topics in fluorescence spectroscopy is available in our digital library an online access to it is set as public so you can download it instantly. Our digital library saves in multiple locations, allowing you to get the most less latency time to download any of our books like this one. Kindly say, the topics in fluorescence spectroscopy is universally compatible with any devices to read.
75 citations
TL;DR: The applications of surfactants in various fields are gaining more attention, which makes full characterization of the surfactant characterization of growing interest as discussed by the authors, and it is fundamental to measure the critical micelle.
Abstract: The applications of surfactants in various fields are gaining more attention, which makes full characterization of surfactants of growing interest. It is fundamental to measure the critical micelle...
24 citations
TL;DR: The best compound identified in this study showed excellent performance in live cell‐labeling experiments and enabled no‐wash fluorogenic imaging on a timescale of seconds.
Abstract: Fluorescent probes that light-up upon reaction with complementary bioorthogonal reagents are superior tools for no-wash fluorogenic bioimaging applications. In this work, a thorough study is presented on a set of seventeen structurally diverse coumarin-tetrazine probes that produce fluorescent dyes with exceptional turn-on ratios when reacted with trans-cyclooctene (TCO) and bicyclononyne (BCN) dienophiles. In general, formation of the fully aromatic pyridazine-containing dyes resulting from the reaction with BCN was found superior in terms of fluorogenicity. However, evaluation of the probes in cellular imaging experiments revealed that other factors, such as reaction kinetics and good cell permeability, prevail over the fluorescence turn-on properties. The best compound identified in this study showed excellent performance in live cell-labeling experiments and enabled no-wash fluorogenic imaging on a timescale of seconds.
17 citations
TL;DR: The shortened FAST is designed, which is composed of only 98 amino acids, the shortest genetically encoded tag among all known fluorescent and fluorogen-activating proteins, by truncating 26 N-terminal residues.
Abstract: One of the essential characteristics of any tag used in bioscience and medical applications is its size. The larger the label, the more it may affect the studied object, and the more it may distort its behavior. In this paper, using NMR spectroscopy and X-ray crystallography, we have studied the structure of fluorogen-activating protein FAST both in the apo form and in complex with the fluorogen. We showed that significant change in the protein occurs upon interaction with the ligand. While the protein is completely ordered in the complex, its apo form is characterized by higher mobility and disordering of its N-terminus. We used structural information to design the shortened FAST (which we named nanoFAST) by truncating 26 N-terminal residues. Thus, we created the shortest genetically encoded tag among all known fluorescent and fluorogen-activating proteins, which is composed of only 98 amino acids.
17 citations
TL;DR: A critical account of the entire work in the design of chelators to address Mycobacterium avium infections is given and the statement “to label means to change” is justified.
Abstract: Controlling the sources of Fe available to pathogens is one of the possible strategies that can be successfully used by novel antibacterial drugs. We focused our interest on the design of chelators to address Mycobacterium avium infections. Taking into account the molecular structure of mycobacterial siderophores and considering that new chelators must be able to compete for Fe(III), we selected ligands of the 3-hydroxy-4-pyridinone class to achieve our purpose. After choosing the type of chelating unit it was also our objective to design chelators that could be monitored inside the cell and for that reason we designed chelators that could be functionalized with fluorophores. We didn’t realize at the time that the incorporation a fluorophore, to allow spectroscopic detection, would be so relevant for the antimycobacterial effect or to determine the affinity of the chelators towards biological membranes. From a biophysical perspective, this is a fascinating illustration of the fact that functionalization of a molecule with a particular label may lead to a change in its membrane permeation properties and result in a dramatic change in biological activity. For that reason we believe it is interesting to give a critical account of our entire work in this area and justify the statement “to label means to change”. New perspectives regarding combined therapeutic approaches and the use of rhodamine B conjugates to target closely related problems such as bacterial resistance and biofilm production are also discussed.
10 citations
References
More filters
Book•
01 Jan 1983
TL;DR: This book describes the fundamental aspects of fluorescence, the biochemical applications of this methodology, and the instrumentation used in fluorescence spectroscopy.
Abstract: Fluorescence methods are being used increasingly in biochemical, medical, and chemical research. This is because of the inherent sensitivity of this technique. and the favorable time scale of the phenomenon of fluorescence. 8 Fluorescence emission occurs about 10- sec (10 nsec) after light absorp tion. During this period of time a wide range of molecular processes can occur, and these can effect the spectral characteristics of the fluorescent compound. This combination of sensitivity and a favorable time scale allows fluorescence methods to be generally useful for studies of proteins and membranes and their interactions with other macromolecules. This book describes the fundamental aspects of fluorescence. and the biochemical applications of this methodology. Each chapter starts with the -theoreticalbasis of each phenomenon of fluorescence, followed by examples which illustrate the use of the phenomenon in the study of biochemical problems. The book contains numerous figures. It is felt that such graphical presentations contribute to pleasurable reading and increased understand ing. Separate chapters are devoted to fluorescence polarization, lifetimes, quenching, energy transfer, solvent effects, and excited state reactions. To enhance the usefulness of this work as a textbook, problems are included which illustrate the concepts described in each chapter. Furthermore, a separate chapter is devoted to the instrumentation used in fluorescence spectroscopy. This chapter will be especially valuable for those perform ing or contemplating fluorescence measurements. Such measurements are easily compromised by failure to consider a number of simple principles."
28,073 citations
"Fluorophore tagged bio-molecules an..." refers background in this paper
...Fluorophores are usually classified in two categories; intrinsic and extrinsic (1, 18)....
[...]
25 Apr 2012
TL;DR: In this article, the effects of intermolecular photophysical processes on fluorescence emission are discussed and an analysis of the effect of polarity of fluorescence emissions is presented.
Abstract: Preface. Prologue. Introduction. Absorption of UV--visible light. Characteristics of Fluorescence Emission. Effects of Intermolecular Photophysical Processes on Fluorescence Emission. Fluorescence polarization: Emission Ansotropy. Principles of steady--state and time--resolved fluorometric techniques. Effect of polarity of fluorescence emission. Polarity probes. Microviscosity, fluidity, molecular mobility. Estimation by means of fluorescent probes. Resonance energy transfer and its applications. Fluorescent molecular sensors of ions and molecules. Advanced techniques in fluorescence spectroscopy. Epilogue. Index.
4,261 citations
1,859 citations
TL;DR: The lifetime of a photophysical process is the time required by a population of N electronically excited molecules to be reduced by a factor of e via the loss of energy through fluorescence and other non-radiative processes and the average length of time τ is called the mean lifetime, or simply lifetime.
Abstract: When a molecule absorbs a photon of appropriate energy, a chain of photophysical events ensues, such as internal conversion or vibrational relaxation (loss of energy in the absence of light emission), fluorescence, intersystem crossing (from singlet state to a triplet state) and phosphorescence, as shown in the Jablonski diagram for organic molecules (Fig. 1). Each of the processes occurs with a certain probability, characterized by decay rate constants (k). It can be shown that the average length of time τ for the set of molecules to decay from one state to another is reciprocally proportional to the rate of decay: τ = 1/k. This average length of time is called the mean lifetime, or simply lifetime. It can also be shown that the lifetime of a photophysical process is the time required by a population of N electronically excited molecules to be reduced by a factor of e. Correspondingly, the fluorescence lifetime is the time required by a population of excited fluorophores to decrease exponentially to N/e via the loss of energy through fluorescence and other non-radiative processes. The lifetime of photophycal processes vary significantly from tens of femotoseconds for internal conversion1,2 to nanoseconds for fluorescence and microseconds or seconds for phosphorescence.1
Open in a separate window
Figure 1
Jablonski diagram and a timescale of photophysical processes for organic molecules.
1,829 citations
TL;DR: An assay for vesicle--vesicle fusion involving resonance energy transfer between N-(7-nitro-2,1,3-benzoxadiazol-4-yl), the energy donor, and rhodamine, the energy acceptor has been developed.
Abstract: An assay for vesicle--vesicle fusion involving resonance energy transfer between N-(7-nitro-2,1,3-benzoxadiazol-4-yl), the energy donor, and rhodamine, the energy acceptor, has been developed. The two fluorophores are coupled to the free amino group of phosphatidylethanolamine to provide analogues which can be incorporated into a lipid vesicle bilayer. When both fluorescent lipids are in phosphatidylserine vesicles at appropriate surface densities (ratio of fluorescent lipid to total lipid), efficient energy transfer is observed. When such vesicles are fused with a population of pure phosphatidylserine vesicles by the addition of calcium, the two probes mix with the other lipids present to form a new membrane. This mixing reduces the surface density of the energy acceptor resulting in a decreased efficiency of resonance energy transfer which is measured experimentally. These changes in transfer efficiency allow kinetic and quantitative measurements of the fusion process. Using this system, we have studied the ability of phosphatidylcholine, phosphatidylserine, and phosphatidylcholine--phosphatidylserine (1:1) vesicles to fuse with cultured fibroblasts. Under the conditions employed, the majority of the cellular uptake of vesicle lipid could be attributed to the adsorption of intact vesicles to the cell surface regardless of the composition of the vesicle bilayer.
1,262 citations
"Fluorophore tagged bio-molecules an..." refers background or methods in this paper
...With the help of these RET assays distinction between adsorption and fusion is also possible (82)....
[...]
...Lipid NBD, Dansyl Membrane polarity (68, 72) NBD Packing density (65) Pyrene Domain formation (47) NBD Lateral heterogeneity (48) Pentaene Nano-domain formation (73) Pyrene Lipid-protein interaction (74, 75) Fluorescein – BODIPYCoumarin – BODIPY Lipid-protein interaction (76, 77) Pyrene Fusion (61, 78, 79, 80) NBD – Rhodamine Fluorescein – Rhodamine Fluorescein – BODIPY Dansyl – BODIPY Pyrene – BODIPY DPH – BODIPY Coumarin – BODIPY Coumarin – Rhodamine Fusion (61, 78, 81, 82)...
[...]