Submit to IJMS Review for IJMS Propose a Special Issue

Journal Menu

Journal Browser

Advanced Fluorescence Methodologies: Focus on Molecular Research

Print Special Issue Flyer
Special Issue Editors
Special Issue Information
Keywords
Related Special Issue
Published Papers

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Macromolecules".

Deadline for manuscript submissions: closed (30 December 2022) | Viewed by 29504

Share This Special Issue

Special Issue Editors

Prof. Dr. Joseph P. Albanesi

E-Mail Website
Guest Editor

Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
Interests: dynamin; phosphoinositides in membrane trafficking; mechanisms of endocytosis and autophagy
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. David M. Jameson

E-Mail Website
Guest Editor

Department of Cell and Molecular Biology, University of Hawaii at Manoa, 651 Ilalo St., BSB222, Honolulu, HI 96813, USA
Interests: fluorescence methods; FLIM/FRET; protein interactions; FCS; polarization/anisotropy
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Detailed analysis of macromolecular structure and dynamics has been greatly facilitated by the use of advanced fluorescence-based experimental approaches. Although the basic principles of fluorescence spectroscopy and imaging were established decades ago, recent conceptual and technological advances have dramatically extended their range of applicability. For example, improvements in optics, electronics, computation, and dye technology have allowed unprecedented access to macromolecular motions and interactions in vitro and in living cells.

This Special Issue will present original research investigations that employ state-of-the-art fluorescence methodologies, as well as review articles that address current progress in the field. Emphasis will be on studies which advance our understanding of biological systems.

Potential topics include, but are not limited to, the following:

Fluorescence fluctuation spectroscopy
Single-particle tracking
Phasor analysis
Single-molecule analysis
Super-resolution
FRET/FLIM
Fluorescent probes
FRAP

Prof. Dr. Joseph P. Albanesi
Dr. David M. Jameson
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

fluorescence
FRET
FLIM
FRAP
phasors
FCS
single molecule
super-resolution

Related Special Issue

Advanced Fluorescence Methodologies: Focus on Macromolecules Research 2.0 in International Journal of Molecular Sciences (1 article)

Published Papers (13 papers)

Download All Papers

Research

Jump to: Review

17 pages, 7756 KiB

Open AccessArticle

A Novel Indole Derivative with Superior Photophysical Performance for Fluorescent Probe, pH-Sensing, and Logic Gates

by Hai-Ling Liu, Kan Zhan, Kai-Liang Zhong, Xing-Liang Chen and Xing-Hua Xia

Int. J. Mol. Sci. 2023, 24(2), 1711; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms24021711 - 15 Jan 2023

Cited by 1 | Viewed by 1638

Abstract

An indole-related molecules have been considered as the potential fluorescent probes for biological and electrochemical sensing. However, most of the indole probes have been usually used in a single detection mode. Indolium probes that enable accurate detection in complex environments are rarely reported. Here, four novel indole derivatives including the phenyl group substituted with different functional moieties were designed on the basis of the donor-π-acceptor (D-π-A) concept. These derivatives exhibit positive solvatochromism owing to their varied molecular conformations upon contacting to various solvents and the different HOMO-LUMO gaps caused by the difference in electronic push-pull capability of the substituents. Their solid-state fluorescence emissions and multiple chromisms are observed due to the inherent twisted geometries and aggregation modes. In addition, these derivatives show dramatic color and fluorescence responses due to the protonation of the nitrogen and oxygen containing groups, and thus novel colorimetric pH sensors, fluorescent papers and logic gates have been designed. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Graphical abstract

14 pages, 2863 KiB

Open AccessArticle

Probing Red Blood Cell Membrane Microviscosity Using Fluorescence Anisotropy Decay Curves of the Lipophilic Dye PKH26

by Alexey N. Semenov, Daniil A. Gvozdev, Anastasia M. Moysenovich, Dmitry V. Zlenko, Evgenia Yu. Parshina, Adil A. Baizhumanov, Gleb S. Budylin and Eugene G. Maksimov

Int. J. Mol. Sci. 2022, 23(24), 15767; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232415767 - 12 Dec 2022

Cited by 2 | Viewed by 1693

Abstract

Red blood cell (RBC) aggregation and deformation are governed by the molecular processes occurring on the membrane. Since several social important diseases are accompanied by alterations in RBC aggregation and deformability, it is important to develop a diagnostic parameter of RBC membrane structural integrity and stability. In this work, we propose membrane microviscosity assessed by time-resolved fluorescence anisotropy of the lipophilic PKH26 fluorescent probe as a diagnostic parameter. We measured the fluorescence decay curves of the PKH26 probe in the RBC membrane to establish the optimal parameters of the developed fluorescence assay. We observed a complex biphasic profile of the fluorescence anisotropy decay characterized by two correlation times corresponding to the rotational diffusion of free PKH26, and membrane-bounded molecules of the probe. The developed assay allowed us to estimate membrane microviscosity

η_{m}

in the range of 100–500 cP depending on the temperature, which paves the way for assessing RBC membrane properties in clinical applications as predictors of blood microrheological abnormalities. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

13 pages, 3355 KiB

Open AccessArticle

Tumor Targeting with Methotrexate-Conjugated Zwitterionic Near-Infrared Fluorophore for Precise Photothermal Therapy

by Gayoung Jo, Eun Jeong Kim, Min Ho Park and Hoon Hyun

Int. J. Mol. Sci. 2022, 23(22), 14127; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232214127 - 16 Nov 2022

Cited by 3 | Viewed by 1381

Abstract

Targeted tumor imaging can effectively enable image-guided surgery and precise cancer therapy. Finding the right combination of anticancer drugs and near-infrared (NIR) fluorophores is the key to targeted photothermal cancer treatment. In this study, a tumor-targetable NIR fluorophore conjugate with rapid body clearance was developed for accurate tumor imaging and effective photothermal therapy (PTT). The methotrexate (MTX) and zwitterionic NIR fluorophore conjugate (MTX-ZW) were prepared by conjugating a folate antagonist MTX with an aminated ZW800-1 analog to increase the tumor targetability for NIR laser-based PTT of cancer. The MTX, known as a poor tumor-selective drug, showed high tumor accumulation and rapid background clearance after conjugation with the highly water-soluble zwitterionic NIR fluorophore up to 4 h post-injection. The photothermal energy was generated from the MTX-ZW conjugate to induce necrotic cell death in the targeted tumor site under 808 nm laser irradiation. Compared with the previously reported MTX conjugates, the MTX-ZW conjugate can be a great candidate for targeted tumor imaging and fluorescence-guided photothermal cancer therapy. Therefore, these results provide a strategy for the design of drug-fluorophore conjugates and elaborate therapeutic platforms for cancer phototherapy. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

11 pages, 1749 KiB

Open AccessArticle

A Single Fluorescent Protein-Based Indicator with a Time-Resolved Fluorescence Readout for Precise pH Measurements in the Alkaline Range

by Tatiana R. Simonyan, Elena A. Protasova, Anastasia V. Mamontova, Aleksander M. Shakhov, Konstantin A. Lukyanov, Eugene G. Maksimov and Alexey M. Bogdanov

Int. J. Mol. Sci. 2022, 23(21), 12907; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232112907 - 26 Oct 2022

Cited by 2 | Viewed by 1662

Abstract

The real-time monitoring of the intracellular pH in live cells with high precision represents an important methodological challenge. Although genetically encoded fluorescent indicators can be considered as a probe of choice for such measurements, they are hindered mostly by the inability to determine an absolute pH value and/or a narrow dynamic range of the signal, making them inefficient for recording the small pH changes that typically occur within cellular organelles. Here, we study the pH sensitivity of a green-fluorescence-protein (GFP)-based emitter (EGFP-Y145L/S205V) with the alkaline-shifted chromophore’s pKa and demonstrate that, in the pH range of 7.5–9.0, its fluorescence lifetime changes by a factor of ~3.5 in a quasi-linear manner in mammalian cells. Considering the relatively strong lifetime response in a narrow pH range, we proposed the mitochondria, which are known to have a weakly alkaline milieu, as a target for live-cell pH measurements. Using fluorescence lifetime imaging microscopy (FLIM) to visualize the HEK293T cells expressing mitochondrially targeted EGFP-Y145L/S205V, we succeeded in determining the absolute pH value of the mitochondria and recorded the ETC-uncoupler-stimulated pH shift with a precision of 0.1 unit. We thus show that a single GFP with alkaline-shifted pKa can act as a high-precision indicator that can be used in a specific pH range. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

19 pages, 2784 KiB

Open AccessArticle

Computational Modeling and Imaging of the Intracellular Oxygen Gradient

by Andrew J. H. Sedlack, Rozhin Penjweini, Katie A. Link, Alexandra Brown, Jeonghan Kim, Sung-Jun Park, Jay H. Chung, Nicole Y. Morgan and Jay R. Knutson

Int. J. Mol. Sci. 2022, 23(20), 12597; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232012597 - 20 Oct 2022

Cited by 2 | Viewed by 2340

Abstract

Computational modeling can provide a mechanistic and quantitative framework for describing intracellular spatial heterogeneity of solutes such as oxygen partial pressure (pO₂). This study develops and evaluates a finite-element model of oxygen-consuming mitochondrial bioenergetics using the COMSOL Multiphysics program. The model derives steady-state oxygen (O₂) distributions from Fickian diffusion and Michaelis–Menten consumption kinetics in the mitochondria and cytoplasm. Intrinsic model parameters such as diffusivity and maximum consumption rate were estimated from previously published values for isolated and intact mitochondria. The model was compared with experimental data collected for the intracellular and mitochondrial pO₂ levels in human cervical cancer cells (HeLa) in different respiratory states and under different levels of imposed pO₂. Experimental pO₂ gradients were measured using lifetime imaging of a Förster resonance energy transfer (FRET)-based O₂ sensor, Myoglobin-mCherry, which offers in situ real-time and noninvasive measurements of subcellular pO₂ in living cells. On the basis of these results, the model qualitatively predicted (1) the integrated experimental data from mitochondria under diverse experimental conditions, and (2) the impact of changes in one or more mitochondrial processes on overall bioenergetics. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

22 pages, 5444 KiB

Open AccessArticle

Mapping the Nicotinic Acetylcholine Receptor Nanocluster Topography at the Cell Membrane with STED and STORM Nanoscopies

by Lucas A. Saavedra, Héctor Buena-Maizón and Francisco J. Barrantes

Int. J. Mol. Sci. 2022, 23(18), 10435; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms231810435 - 09 Sep 2022

Cited by 1 | Viewed by 1989

Abstract

The cell-surface topography and density of nicotinic acetylcholine receptors (nAChRs) play a key functional role in the synapse. Here we employ in parallel two labeling and two super-resolution microscopy strategies to characterize the distribution of this receptor at the plasma membrane of the mammalian clonal cell line CHO-K1/A5. Cells were interrogated with two targeted techniques (confocal microscopy and stimulated emission depletion (STED) nanoscopy) and single-molecule nanoscopy (stochastic optical reconstruction microscopy, STORM) using the same fluorophore, Alexa Fluor 647, tagged onto either α-bungarotoxin (BTX) or the monoclonal antibody mAb35. Analysis of the topography of nanometer-sized aggregates (“nanoclusters”) was carried out using STORMGraph, a quantitative clustering analysis for single-molecule localization microscopy based on graph theory and community detection, and ASTRICS, an inter-cluster similarity algorithm based on computational geometry. Antibody-induced crosslinking of receptors resulted in nanoclusters with a larger number of receptor molecules and higher densities than those observed in BTX-labeled samples. STORM and STED provided complementary information, STED rendering a direct map of the mesoscale nAChR distribution at distances ~10-times larger than the nanocluster centroid distances measured in STORM samples. By applying photon threshold filtering analysis, we show that it is also possible to detect the mesoscale organization in STORM images. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

9 pages, 1563 KiB

Open AccessArticle

Breaking the Concentration Limit in Fluorescence Fluctuation Spectroscopy with Camera-Based Detection

by Yu-Kai Huang and Per Niklas Hedde

Int. J. Mol. Sci. 2022, 23(17), 9840; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23179840 - 30 Aug 2022

Viewed by 1412

Abstract

Fluorescence correlation spectroscopy (FCS) is an extremely versatile tool that has been widely used to measure chemical reaction rates, protein binding, nanoparticle-protein interactions, and biomolecular dynamics in vitro and in vivo. As an inherently micro-sized approach, FCS is compatible with high-throughput screening applications, as demanded for drug design, but typically limited to nanomolar concentrations, which restricts possible applications. Here, we show how massively parallel camera-based detection with side illumination can extend the usable concentration range of FCS more than 100-fold to measure low affinity processes. Our line illumination (LIM) approach is robust, fast (1 s acquisition times), and does not require any reference measurements to characterize the observation volume size. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

16 pages, 2271 KiB

Open AccessArticle

Measuring Molecular Diffusion in Dynamic Subcellular Nanostructures by Fast Raster Image Correlation Spectroscopy and 3D Orbital Tracking

by Filippo Begarani, Francesca D’Autilia, Gianmarco Ferri, Luca Pesce, Fabio Azzarello, Valentina De Lorenzi, William Durso, Ambra Del Grosso, Marco Cecchini and Francesco Cardarelli

Int. J. Mol. Sci. 2022, 23(14), 7623; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23147623 - 10 Jul 2022

Cited by 1 | Viewed by 1690

Abstract

Here we provide demonstration that fast fluorescence fluctuation spectroscopy is a fast and robust approach to extract information on the dynamics of molecules enclosed within subcellular nanostructures (e.g., organelles or vesicles) which are also moving in the complex cellular environment. In more detail, Raster Image Correlation Spectroscopy (RICS) performed at fast timescales (i.e., microseconds) reveals the fast motion of fluorescently labeled molecules within two exemplary dynamic subcellular nanostructures of biomedical interest, the lysosome and the insulin secretory granule (ISG). The measurement of molecular diffusion is then used to extract information on the average properties of subcellular nanostructures, such as macromolecular crowding or molecular aggregation. Concerning the lysosome, fast RICS on a fluorescent tracer allowed us to quantitatively assess the increase in organelle viscosity in the pathological condition of Krabbe disease. In the case of ISGs, fast RICS on two ISG-specific secreting peptides unveiled their differential aggregation propensity depending on intragranular concentration. Finally, a combination of fast RICS and feedback-based 3D orbital tracking was used to subtract the slow movement of subcellular nanostructures from the fast diffusion of molecules contained within them and independently validate the results. Results presented here not only demonstrate the acquired ability to address the dynamic behavior of molecules in moving, nanoscopic reference systems, but prove the relevance of this approach to advance our knowledge on cell function at the subcellular scale. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

14 pages, 2649 KiB

Open AccessArticle

Fluorescence Lifetime Phasor Analysis of the Decamer–Dimer Equilibrium of Human Peroxiredoxin 1

by Sebastián F. Villar, Joaquín Dalla-Rizza, Matías N. Möller, Gerardo Ferrer-Sueta, Leonel Malacrida, David M. Jameson and Ana Denicola

Int. J. Mol. Sci. 2022, 23(9), 5260; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23095260 - 09 May 2022

Cited by 5 | Viewed by 3599

Abstract

Protein self-assembly is a common feature in biology and is often required for a myriad of fundamental processes, such as enzyme activity, signal transduction, and transport of solutes across membranes, among others. There are several techniques to find and assess homo-oligomer formation in proteins. Naturally, all these methods have their limitations, meaning that at least two or more different approaches are needed to characterize a case study. Herein, we present a new method to study protein associations using intrinsic fluorescence lifetime with phasors. In this case, the method is applied to determine the equilibrium dissociation constant (K_D) of human peroxiredoxin 1 (hPrx1), an efficient cysteine-dependent peroxidase, that has a quaternary structure comprised of five head-to-tail homodimers non-covalently arranged in a decamer. The hPrx1 oligomeric state not only affects its activity but also its association with other proteins. The excited state lifetime of hPrx1 has distinct values at high and low concentrations, suggesting the presence of two different species. Phasor analysis of hPrx1 emission lifetime allowed for the identification and quantification of hPrx1 decamers, dimers, and their mixture at diverse protein concentrations. Using phasor algebra, we calculated the fraction of hPrx1 decamers at different concentrations and obtained K_D (1.1 × 10⁻²⁴ M⁴) and C_0.5 (1.36 μM) values for the decamer–dimer equilibrium. The results were validated and compared with size exclusion chromatography. In addition, spectral phasors provided similar results despite the small differences in emission spectra as a function of hPrx1 concentration. The phasor approach was shown to be a highly sensitive and quantitative method to assess protein oligomerization and an attractive addition to the biophysicist’s toolkit. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

10 pages, 6321 KiB

Open AccessArticle

Add and Go: FRET Acceptor for Live-Cell Measurements Modulated by Externally Provided Ligand

by Alexey S. Gavrikov, Nina G. Bozhanova, Mikhail S. Baranov and Alexander S. Mishin

Int. J. Mol. Sci. 2022, 23(8), 4396; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23084396 - 15 Apr 2022

Viewed by 1629

Abstract

A substantial number of genetically encoded fluorescent sensors rely on the changes in FRET efficiency between fluorescent cores, measured in ratiometric mode, with acceptor photobleaching or by changes in fluorescence lifetime. We report on a modulated FRET acceptor allowing for simplified one-channel FRET measurement based on a previously reported fluorogen-activating protein, DiB1. Upon the addition of the cell-permeable chromophore, the fluorescence of the donor-fluorescent protein mNeonGreen decreases, allowing for a simplified one-channel FRET measurement. The reported chemically modulated FRET acceptor is compatible with live-cell experiments and allows for prolonged time-lapse experiments with dynamic energy transfer evaluation. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

17 pages, 7040 KiB

Open AccessArticle

Optical Study of Solvatochromic Isocyanoaminoanthracene Dyes and 1,5-Diaminoanthracene

by Miklós Nagy, Béla Fiser, Milán Szőri, László Vanyorek and Béla Viskolcz

Int. J. Mol. Sci. 2022, 23(3), 1315; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23031315 - 24 Jan 2022

Cited by 4 | Viewed by 3227

Abstract

Isocyanoaminoarenes (ICAAr-s) are a novel and versatile group of solvatochromic fluorophores. Despite their versatile applicability, such as antifungals, cancer drugs and analytical probes, they still represent a mostly unchartered territory among intramolecular charge-transfer (ICT) dyes. The current paper describes the preparation and detailed optical study of novel 1-isocyano-5-aminoanthrace (ICAA) and its N-methylated derivatives along with the starting 1,5-diaminoanthracene. The conversion of one of the amino groups of the diamine into an isocyano group significantly increased the polar character of the dyes, which resulted in a significant 50–70 nm (2077–2609 cm⁻¹) redshift of the emission maximum and a broadened solvatochromic range. The fluorescence quantum yield of ICAAs is strongly influenced by the polarity of the solvent. The starting anthracene-diamine is highly fluorescent in every solvent (√_f = 12–53%), while the isocyano derivatives are practically nonfluorescent in solvents more polar than dioxane. This phenomenon implies the potential application of ICAAs to probe the polarity of the medium and is favorable in practical applications, such as cell-staining, resulting in a reduced background fluorescence. The ICT character of the emission states of ICAAs are in good agreement with the computational findings presented in TD-DFT calculations and molecular electrostatic potential (MESP) isosurfaces. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

Review

Jump to: Research

37 pages, 4183 KiB

Open AccessReview

Choosing the Probe for Single-Molecule Fluorescence Microscopy

by Chiara Schirripa Spagnolo and Stefano Luin

Int. J. Mol. Sci. 2022, 23(23), 14949; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232314949 - 29 Nov 2022

Cited by 7 | Viewed by 3091

Abstract

Probe choice in single-molecule microscopy requires deeper evaluations than those adopted for less sensitive fluorescence microscopy studies. Indeed, fluorophore characteristics can alter or hide subtle phenomena observable at the single-molecule level, wasting the potential of the sophisticated instrumentation and algorithms developed for advanced single-molecule applications. There are different reasons for this, linked, e.g., to fluorophore aspecific interactions, brightness, photostability, blinking, and emission and excitation spectra. In particular, these spectra and the excitation source are interdependent, and the latter affects the autofluorescence of sample substrate, medium, and/or biological specimen. Here, we review these and other critical points for fluorophore selection in single-molecule microscopy. We also describe the possible kinds of fluorophores and the microscopy techniques based on single-molecule fluorescence. We explain the importance and impact of the various issues in fluorophore choice, and discuss how this can become more effective and decisive for increasingly demanding experiments in single- and multiple-color applications. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

18 pages, 3172 KiB

Open AccessReview

Studying Chromatin Epigenetics with Fluorescence Microscopy

by Afanasii I. Stepanov, Zlata V. Besedovskaia, Maria A. Moshareva, Konstantin A. Lukyanov and Lidia V. Putlyaeva

Int. J. Mol. Sci. 2022, 23(16), 8988; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23168988 - 12 Aug 2022

Cited by 4 | Viewed by 3152

Abstract

Epigenetic modifications of histones (methylation, acetylation, phosphorylation, etc.) are of great importance in determining the functional state of chromatin. Changes in epigenome underlay all basic biological processes, such as cell division, differentiation, aging, and cancerous transformation. Post-translational histone modifications are mainly studied by immunoprecipitation with high-throughput sequencing (ChIP-Seq). It enables an accurate profiling of target modifications along the genome, but suffers from the high cost of analysis and the inability to work with living cells. Fluorescence microscopy represents an attractive complementary approach to characterize epigenetics. It can be applied to both live and fixed cells, easily compatible with high-throughput screening, and provide access to rich spatial information down to the single cell level. In this review, we discuss various fluorescent probes for histone modification detection. Various types of live-cell imaging epigenetic sensors suitable for conventional as well as super-resolution fluorescence microscopy are described. We also focus on problems and future perspectives in the development of fluorescent probes for epigenetics. Full article

(This article belongs to the Special Issue Advanced Fluorescence Methodologies: Focus on Molecular Research)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Advanced Fluorescence Methodologies: Focus on Molecular Research

Share This Special Issue

Special Issue Editors

Special Issue Information

Keywords

Related Special Issue

Published Papers (13 papers)

Research

Review

Further Information

Guidelines

MDPI Initiatives

Follow MDPI