Special Issue "Fluorescent Probes for Live Cell Imaging"

A special issue of Chemosensors (ISSN 2227-9040).

Deadline for manuscript submissions: closed (31 August 2018)

Special Issue Editors

Guest Editor
Dr. Zhijie Chen

University of California, Berkeley, CA, United States
Website | E-Mail
Interests: genetically encoded fluorescent probes; hydrogen sulfide; peroxynitrite; zinc; redox biology; unnatural amino acids; live cell imaging; enzyme catalysis; single molecule biophysics; optical tweezers; transcription; epigenetics
Guest Editor
Dr. Grazvydas Lukinavicius

Max Planck Institute for Biophysical Chemistry, Gottingen, Niedersachsen, Germany
Website | E-Mail
Interests: chemical biology; fluorescent probes; super-resolution microscopy; living cell imaging

Special Issue Information

Dear Colleagues,

Fluorescent probes are essential molecular tools for the detection and imaging of biological molecules. The availability of robust probes for a particular molecular species often catalyzes unprecedented advancements of an entire field, as exemplified by genetically encoded calcium indicators. In order to understand cell physiology at a molecular level, we need fluorescent probes for each cellular component such as metal ions, metabolites, lipids, proteins and nucleic acids. These fluorescent probes when coupled with live cell imaging, empower the study of biological molecules noninvasively and in its native environment.

Over the last decade, many new fluorescence imaging methods have emerged allowing for the observation of whole body processes and molecular processes in living organisms. For example, techniques in whole body and thick tissue fluorescence imaging are quickly expanding because of their potential surgical applications. Camera image-guided techniques augment surgeons’ vision. The miniaturization of flow cytometry devices has brought fluorescence imaging to the point-of-care level. Finally, super-resolution microscopy, or nanoscopy, allows objects to be observed at the nanometre (nm, 10−9 m) scale at resolutions that are only slightly lower than with electron microscopy. The importance of this discovery is emphasized by 2014 Nobel Prize in Chemistry award.

All above mentioned techniques rely on and facilitate the development of the fluorescent probes and sensors. A special attention is payed to the molecules compatible with living cell imaging allowing vizualization of dynamics, transformations and localization. This Special Issue is focusing on such fluorescent probes and attempts to build a roadmap, which, ideally, should provide useful guidelines for future developments in this vast and dynamic field.

Dr. Zhijie Chen
Dr. Grazvydas Lukinavicius
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 papers will be 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. Chemosensors is an international peer-reviewed open access quarterly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 350 CHF (Swiss Francs). 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

  • Fluorescent probes
  • Fluorescence imaging
  • Molecular imaging
  • Sensors
  • Biosensors
  • Fluorescent indicators
  • Live cell imaging
  • Whole body and thick tissue fluorescence imaging
  • Camera image-guided surgery
  • Super-resolution microscopy
  • Metabolites
  • Small molecules
  • Genetically encoded fluorescent probes
  • Chemical probes
  • Cell signaling

Published Papers (7 papers)

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Editorial

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Open AccessEditorial Fluorescent Probes for Live Cell Imaging
Chemosensors 2018, 6(3), 41; https://doi.org/10.3390/chemosensors6030041
Received: 18 September 2018 / Accepted: 19 September 2018 / Published: 19 September 2018
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(This article belongs to the Special Issue Fluorescent Probes for Live Cell Imaging)

Research

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Open AccessFeature PaperArticle Using Fluorescence Intensity of Enhanced Green Fluorescent Protein to Quantify Pseudomonas aeruginosa
Chemosensors 2018, 6(2), 21; https://doi.org/10.3390/chemosensors6020021
Received: 30 March 2018 / Revised: 25 April 2018 / Accepted: 27 April 2018 / Published: 3 May 2018
Cited by 1 | PDF Full-text (1772 KB) | HTML Full-text | XML Full-text
Abstract
A variety of direct and indirect methods have been used to quantify planktonic and biofilm bacterial cells. Direct counting methods to determine the total number of cells include plate counts, microscopic cell counts, Coulter cell counting, flow cytometry, and fluorescence microscopy. However, indirect
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A variety of direct and indirect methods have been used to quantify planktonic and biofilm bacterial cells. Direct counting methods to determine the total number of cells include plate counts, microscopic cell counts, Coulter cell counting, flow cytometry, and fluorescence microscopy. However, indirect methods are often used to supplement direct cell counting, as they are often more convenient, less time-consuming, and require less material, while providing a number that can be related to the direct cell count. Herein, an indirect method is presented that uses fluorescence emission intensity as a proxy marker for studying bacterial accumulation. A clinical strain of Pseudomonas aeruginosa was genetically modified to express a green fluorescent protein (PA14/EGFP). The fluorescence intensity of EGFP in live cells was used as an indirect measure of live cell density, and was compared with the traditional cell counting methods of optical density (OD600) and plate counting (colony-forming units (CFUs)). While both OD600 and CFUs are well-established methods, the use of fluorescence spectroscopy to quantify bacteria is less common. This study demonstrates that EGFP intensity is a convenient reporter for bacterial quantification. In addition, we demonstrate the potential for fluorescence spectroscopy to be used to measure the quantity of PA14/EGFP biofilms, which have important human health implications due to their antimicrobial resistance. Therefore, fluorescence spectroscopy could serve as an alternative or complementary quick assay to quantify bacteria in planktonic cultures and biofilms. Full article
(This article belongs to the Special Issue Fluorescent Probes for Live Cell Imaging)
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Open AccessFeature PaperArticle A Rationally Designed, Spiropyran-Based Chemosensor for Magnesium
Chemosensors 2018, 6(2), 17; https://doi.org/10.3390/chemosensors6020017
Received: 8 March 2018 / Revised: 14 April 2018 / Accepted: 16 April 2018 / Published: 17 April 2018
Cited by 1 | PDF Full-text (12420 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Magnesium ions (Mg2+) play an important role in mammalian cell function; however, relatively little is known about the mechanisms of Mg2+ regulation in disease states. An advance in this field would come from the development of selective, reversible fluorescent chemosensors,
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Magnesium ions (Mg2+) play an important role in mammalian cell function; however, relatively little is known about the mechanisms of Mg2+ regulation in disease states. An advance in this field would come from the development of selective, reversible fluorescent chemosensors, capable of repeated measurements. To this end, the rational design and fluorescence-based photophysical characterisation of two spiropyran-based chemosensors for Mg2+ are presented. The most promising analogue, chemosensor 1, exhibits 2-fold fluorescence enhancement factor and 3-fold higher binding affinity for Mg2+ (Kd 6.0 µM) over Ca2+ (Kd 18.7 µM). Incorporation of spiropyran-based sensors into optical fibre sensing platforms has been shown to yield significant signal-to-background changes with minimal sample volumes, a real advance in biological sensing that enables measurement on subcellular-scale samples. In order to demonstrate chemosensor compatibility within the light intense microenvironment of an optical fibre, photoswitching and photostability of 1 within a suspended core optical fibre (SCF) was subsequently explored, revealing reversible Mg2+ binding with improved photostability compared to the non-photoswitchable Rhodamine B fluorophore. The spiropyran-based chemosensors reported here highlight untapped opportunities for a new class of photoswitchable Mg2+ probe and present a first step in the development of a light-controlled, reversible dip-sensor for Mg2+. Full article
(This article belongs to the Special Issue Fluorescent Probes for Live Cell Imaging)
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Review

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Open AccessReview “Probe, Sample, and Instrument (PSI)”: The Hat-Trick for Fluorescence Live Cell Imaging
Chemosensors 2018, 6(3), 40; https://doi.org/10.3390/chemosensors6030040
Received: 14 June 2018 / Revised: 23 August 2018 / Accepted: 10 September 2018 / Published: 13 September 2018
Cited by 1 | PDF Full-text (7136 KB) | HTML Full-text | XML Full-text
Abstract
Cell Imaging Platforms (CIPs) are research infrastructures offering support to a number of scientific projects including the choice of adapted fluorescent probes for live cell imaging. What to detect in what type of sample and for how long is a major issue with
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Cell Imaging Platforms (CIPs) are research infrastructures offering support to a number of scientific projects including the choice of adapted fluorescent probes for live cell imaging. What to detect in what type of sample and for how long is a major issue with fluorescent probes and, for this, the “hat-trick” “Probe–Sample–Instrument” (PSI) has to be considered. We propose here to deal with key points usually discussed in CIPs including the properties of fluorescent organic probes, the modality of cell labeling, and the best equipment to obtain appropriate spectral, spatial, and temporal resolution. New strategies in organic synthesis and click chemistry for accessing probes with enhanced photophysical characteristics and targeting abilities will also be addressed. Finally, methods for image processing will be described to optimize exploitation of fluorescence signals. Full article
(This article belongs to the Special Issue Fluorescent Probes for Live Cell Imaging)
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Open AccessFeature PaperReview Dissection of Protein Kinase Pathways in Live Cells Using Photoluminescent Probes: Surveillance or Interrogation?
Chemosensors 2018, 6(2), 19; https://doi.org/10.3390/chemosensors6020019
Received: 31 March 2018 / Revised: 22 April 2018 / Accepted: 23 April 2018 / Published: 25 April 2018
Cited by 1 | PDF Full-text (3194 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Protein kinases catalyze phosphorylation, a small yet crucial modification that affects participation of the substrate proteins in the intracellular signaling pathways. The activity of 538 protein kinases encoded in human genome relies upon spatiotemporally controlled mechanisms, ensuring correct progression of virtually all physiological
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Protein kinases catalyze phosphorylation, a small yet crucial modification that affects participation of the substrate proteins in the intracellular signaling pathways. The activity of 538 protein kinases encoded in human genome relies upon spatiotemporally controlled mechanisms, ensuring correct progression of virtually all physiological processes on the cellular level—from cell division to cell death. The aberrant functioning of protein kinases is linked to a wide spectrum of major health issues including cancer, cardiovascular diseases, neurodegenerative diseases, inflammatory diseases, etc. Hence, significant effort of scientific community has been dedicated to the dissection of protein kinase pathways in their natural milieu. The combination of recent advances in the field of light microscopy, the wide variety of genetically encoded or synthetic photoluminescent scaffolds, and the techniques for intracellular delivery of cargoes has enabled design of a plethora of probes that can report activation of target protein kinases in human live cells. The question remains: how much do we bias intracellular signaling of protein kinases by monitoring it? This review seeks answers to this question by analyzing different classes of probes according to their general structure, mechanism of recognition of biological target, and optical properties necessary for the reporting of intracellular events. Full article
(This article belongs to the Special Issue Fluorescent Probes for Live Cell Imaging)
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Open AccessReview The Use of Hoechst Dyes for DNA Staining and Beyond
Chemosensors 2018, 6(2), 18; https://doi.org/10.3390/chemosensors6020018
Received: 20 March 2018 / Revised: 15 April 2018 / Accepted: 17 April 2018 / Published: 18 April 2018
Cited by 4 | PDF Full-text (17730 KB) | HTML Full-text | XML Full-text
Abstract
Hoechst dyes are among the most popular fluorophores used to stain DNA in living and fixed cells. Moreover, their high affinity and specificity towards DNA make Hoechst dyes excellent targeting moieties, which can be conjugated to various other molecules in order to tether
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Hoechst dyes are among the most popular fluorophores used to stain DNA in living and fixed cells. Moreover, their high affinity and specificity towards DNA make Hoechst dyes excellent targeting moieties, which can be conjugated to various other molecules in order to tether them to DNA. The recent developments in the fields of microscopy and flow cytometry have sparked interest in such composite molecules, whose applications range from investigating nucleus microenvironment to drug delivery into tumours. Here we provide an overview of the properties of Hoechst dyes and discuss recent developments in Hoechst-based composite probes. Full article
(This article belongs to the Special Issue Fluorescent Probes for Live Cell Imaging)
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Open AccessReview Illuminating Brain Activities with Fluorescent Protein-Based Biosensors
Chemosensors 2017, 5(4), 32; https://doi.org/10.3390/chemosensors5040032
Received: 9 October 2017 / Revised: 19 November 2017 / Accepted: 22 November 2017 / Published: 28 November 2017
Cited by 2 | PDF Full-text (2373 KB) | HTML Full-text | XML Full-text
Abstract
Fluorescent protein-based biosensors are indispensable molecular tools for life science research. The invention and development of high-fidelity biosensors for a particular molecule or molecular event often catalyze important scientific breakthroughs. Understanding the structural and functional organization of brain activities remain a subject for
[...] Read more.
Fluorescent protein-based biosensors are indispensable molecular tools for life science research. The invention and development of high-fidelity biosensors for a particular molecule or molecular event often catalyze important scientific breakthroughs. Understanding the structural and functional organization of brain activities remain a subject for which optical sensors are in desperate need and of growing interest. Here, we review genetically encoded fluorescent sensors for imaging neuronal activities with a focus on the design principles and optimizations of various sensors. New bioluminescent sensors useful for deep-tissue imaging are also discussed. By highlighting the protein engineering efforts and experimental applications of these sensors, we can consequently analyze factors influencing their performance. Finally, we remark on how future developments can fill technological gaps and lead to new discoveries. Full article
(This article belongs to the Special Issue Fluorescent Probes for Live Cell Imaging)
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