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Biosensors
Special Issues
Recent Advances and Perspectives of Fluorescent Biosensors
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Recent Advances and Perspectives of Fluorescent Biosensors

A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Biosensor and Bioelectronic Devices".

Deadline for manuscript submissions: 30 September 2025 | Viewed by 7656

Special Issue Editors

Dr. Miho Suzuki

E-Mail Website
Guest Editor
Faculty of Engineering, Department of Applied Chemistry, Saitama University, Saitama, Japan
Interests: live imaging
Dr. Yutaka Shindo

E-Mail Website
Guest Editor
Faculty of Science and Technology, Department of Biosciences and Informatics, Keio University, Tokyo, Japan
Interests: fluorescence imaging

Special Issue Information

Dear Colleagues,

Tools to elucidate biological phenomena have been regularly developed. Especially in the last few decades, fluorescent biosensors as different approaches from omics studies have continued to be developed together with devices employed for quantitative detections of biomolecule dynamics and activities. Though we have several fluorescence measurement methods based on fluorescence intensity, polarization, lifetime, correlation, etc., and exploit them properly, most fluorescent biosensors have been designed and fabricated for target-specific, spatiotemporal resolvable applications in living cells and their signals have been designed as amplifiable to be applied in the biological, medicinal, clinical fields, among others.

Fluorescent biosensors generally consist of fluorescent organic compounds, proteins, nanoparticles and their combinations. Therefore, improvements from various aspects have been made to expand dynamic ranges and enhance selectivity or sensitivity for monitoring. Those trials have enabled researchers to accomplish high-content imaging of cellular signaling and high-throughput screening of agonists or antagonists for objectives.

This Special Issue aims to highlight high-quality results including original research articles and comprehensive reviews in the field of fluorescent biosensors. Articles that focus on or propose new ideas and new directions are particularly welcome.

Dr. Miho Suzuki
Dr. Yutaka Shindo
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. Biosensors is an international peer-reviewed open access monthly 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 2200 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 sensor
  • ratiometric sensor
  • fluorescent protein
  • synthetic probe
  • quantum dot
  • optogenetic reporter
  • genetically encoded probe
  • live imaging
  • FRET
  • FLIM
  • FCS
  • aggregation-induced emission

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Published Papers (4 papers)

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Research

12 pages, 2689 KiB  
Article
Direct Detection and Quantification of Aqueous Proteins via a Fluorescent Probe Through the Use of Fluorophore-Induced Plasmonic Current
by Daniel R. Pierce and Chris D. Geddes
Biosensors 2025, 15(3), 150; https://doi.org/10.3390/bios15030150 - 27 Feb 2025
Viewed by 417
Abstract
We report on the recent advancements in the sensing of proteins, both directly and with the use of a fluorescent probe, through the use of Fluorophore-Induced Plasmonic Current (FIPC). FIPC are a phenomenon where a fluorophore or excited state species is in close [...] Read more.
We report on the recent advancements in the sensing of proteins, both directly and with the use of a fluorescent probe, through the use of Fluorophore-Induced Plasmonic Current (FIPC). FIPC are a phenomenon where a fluorophore or excited state species is in close proximity to a plasmonically active metal nanoparticle film (MNF), and the excited state is able to couple to the particle, ultimately leading to enhanced spectroscopic properties. This phenomenon is similar to the well-reported metal-enhanced fluorescence (MEF) phenomenon, wherein the coupled complex produces an enhanced fluorescence emission and a shorter lifetime. However, if the particles themselves are sufficiently spaced and oriented, an induced current can transfer from each discreet particle to the next, creating a detectable current across the film. This detectable current has a magnitude that is proportional to the fluorescent properties of the species that produced it, and can be affected by the polarization of the excitation source; the spacing and size of the particles on the film; the overlap between the spectral properties of the film and the species; as well as externally applied voltages and currents. In this study, we examined whether it is possible to detect protein species, directly due to both their intrinsic fluorescent and absorptive properties, and how that compares to commercially available protein detection probes, in a similar manner to prior work by our group addressing analyte detection via turn-on fluorescent probes. This FIPC-based detection technique is a novel method that has not been used for the detection of proteins, and the use of this method could expand the dynamic sensing range of first-pass testing, while overcoming some of the physical limitations on the upper limit of detection of both absorption spectroscopy and fluorescence emission spectroscopy. Our experiments sought to highlight the selectivity of FIPC-based detection relative to both fluorescence and absorption spectroscopy, as well as its sensitivity when working with protein analytes. We examined the effects of protein concentration, intrinsic fluorescent properties, and turn-on probes, as well as how these techniques compare to traditional analytical techniques used today. Full article
(This article belongs to the Special Issue Recent Advances and Perspectives of Fluorescent Biosensors)
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17 pages, 4073 KiB  
Article
Fluorescence Polarization Immunoassay for Rapid, Sensitive Detection of the Herbicide 2,4-Dichlorophenoxyacetic Acid in Juice and Water Samples
by Liliya I. Mukhametova, Marya K. Kolokolova, Ivan A. Shevchenko, Boris S. Tupertsev, Anatoly V. Zherdev, Chuanlai Xu and Sergei A. Eremin
Biosensors 2025, 15(1), 32; https://doi.org/10.3390/bios15010032 - 9 Jan 2025
Viewed by 921
Abstract
2,4-Dichlorophenoxyacetic acid (2,4-D) is one of the popular herbicides that is widely used in agriculture and can be found in food and water. A rapid and sensitive fluorescence polarization immunoassay (FPIA) was proposed for the detection of 2,4-D in juice and water. New [...] Read more.
2,4-Dichlorophenoxyacetic acid (2,4-D) is one of the popular herbicides that is widely used in agriculture and can be found in food and water. A rapid and sensitive fluorescence polarization immunoassay (FPIA) was proposed for the detection of 2,4-D in juice and water. New tracers, 2,4-D-buthylenediamin fluoresceinthiocarbamyl (2,4-D-BDF) and 2,4-D-glycine aminofluorescein (2,4-D-GAF), were obtained and characterized. Monoclonal antibodies (MAb) obtained against 2,4-D were used as a recognition reagent. The kinetics of the interaction of MAb and tracers were studied, and the kinetic parameters of their binding were calculated. High specificity of binding of tracers and MAb was shown. In this work, an approach was elaborated on to reduce the detection limit of 2,4-D by the FPIA method by changing the volume of the studied sample. The optimized FPIA in a competitive format was characterized by the LODs of 2,4-D 8 and 0.4 ng/mL and the working ranges 30–3000 ng/mL and 3–300 ng/mL for juice and water, respectively. The entire test cycle (from sample receipt to evaluation of the analysis results) took only 20 min. The test for the recovery of 2,4-D in juice and water gave values from 95 to 120%, which demonstrated the reliability of the herbicide determination in real samples. Full article
(This article belongs to the Special Issue Recent Advances and Perspectives of Fluorescent Biosensors)
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14 pages, 4310 KiB  
Article
Suppression of Contraction Raises Calcium Ion Levels in the Heart of Zebrafish Larvae
by Antonio Martinez-Sielva, Manuel Vicente, Jussep Salgado-Almario, Aarón Garcia-Blazquez, Beatriz Domingo and Juan Llopis
Biosensors 2024, 14(5), 219; https://doi.org/10.3390/bios14050219 - 27 Apr 2024
Cited by 2 | Viewed by 3339
Abstract
Zebrafish larvae have emerged as a valuable model for studying heart physiology and pathophysiology, as well as for drug discovery, in part thanks to its transparency, which simplifies microscopy. However, in fluorescence-based optical mapping, the beating of the heart results in motion artifacts. [...] Read more.
Zebrafish larvae have emerged as a valuable model for studying heart physiology and pathophysiology, as well as for drug discovery, in part thanks to its transparency, which simplifies microscopy. However, in fluorescence-based optical mapping, the beating of the heart results in motion artifacts. Two approaches have been employed to eliminate heart motion during calcium or voltage mapping in zebrafish larvae: the knockdown of cardiac troponin T2A and the use of myosin inhibitors. However, these methods disrupt the mechano-electric and mechano-mechanic coupling mechanisms. We have used ratiometric genetically encoded biosensors to image calcium in the beating heart of intact zebrafish larvae because ratiometric quantification corrects for motion artifacts. In this study, we found that halting heart motion by genetic means (injection of tnnt2a morpholino) or chemical tools (incubation with para-aminoblebbistatin) leads to bradycardia, and increases calcium levels and the size of the calcium transients, likely by abolishing a feedback mechanism that connects contraction with calcium regulation. These outcomes were not influenced by the calcium-binding domain of the gene-encoded biosensors employed, as biosensors with a modified troponin C (Twitch-4), calmodulin (mCyRFP1-GCaMP6f), or the photoprotein aequorin (GFP-aequorin) all yielded similar results. Cardiac contraction appears to be an important regulator of systolic and diastolic Ca2+ levels, and of the heart rate. Full article
(This article belongs to the Special Issue Recent Advances and Perspectives of Fluorescent Biosensors)
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14 pages, 4396 KiB  
Article
Live Cell Monitoring of Separase Activity, a Key Enzymatic Reaction for Chromosome Segregation, with Chimeric FRET-Based Molecular Sensor upon Cell Cycle Progression
by Md. Shazadur Rahman, Yutaka Shindo, Kotaro Oka, Wataru Ikeda and Miho Suzuki
Biosensors 2024, 14(4), 192; https://doi.org/10.3390/bios14040192 - 15 Apr 2024
Viewed by 2290
Abstract
Separase is a key cysteine protease in the separation of sister chromatids through the digestion of the cohesin ring that inhibits chromosome segregation as a trigger of the metaphase–anaphase transition in eukaryotes. Its activity is highly regulated by binding with securin and cyclinB-CDK1 [...] Read more.
Separase is a key cysteine protease in the separation of sister chromatids through the digestion of the cohesin ring that inhibits chromosome segregation as a trigger of the metaphase–anaphase transition in eukaryotes. Its activity is highly regulated by binding with securin and cyclinB-CDK1 complex. These bindings prevent the proteolytic activity of separase until the onset of anaphase. Chromosome missegregation and aneuploidy are frequently observed in malignancies. However, there are some difficulties in biochemical examinations due to the instability of separase in vitro and the fact that few spatiotemporal resolution approaches exist for monitoring live separase activity throughout mitotic processes. Here, we have developed FRET-based molecular sensors, including GFP variants, with separase-cleavable sequences as donors and covalently attached fluorescent dyes as acceptor molecules. These are applicable to conventional live cell imaging and flow cytometric analysis because of efficient live cell uptake. We investigated the performance of equivalent molecular sensors, either localized or not localized inside the nucleus under cell cycle control, using flow cytometry. Synchronized cell cycle progression rendered significant separase activity detections in both molecular sensors. We obtained consistent outcomes with localized molecular sensor introduction and cell cycle control by fluorescent microscopic observations. We thus established live cell separase activity monitoring systems that can be used specifically or statistically, which could lead to the elucidation of separase properties in detail. Full article
(This article belongs to the Special Issue Recent Advances and Perspectives of Fluorescent Biosensors)
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