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Sensors in Fluorescence Imaging
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Sensors in Fluorescence Imaging

A special issue of Sensors (ISSN 1424-8220). This special issue belongs to the section "Chemical Sensors".

Deadline for manuscript submissions: closed (31 March 2022) | Viewed by 15729

Special Issue Editors

Dr. Suman Ranjit

E-Mail Website
Guest Editor
Assistant Professor, Department of Biochemistry and Molecular & Cellular Biology, Georgetown University, Washington, DC 20007, USA
Interests: fluorescence; FLIM; harmonic generation imaging; SHG/THG; metabolism; phasor; fibrosis
Dr. Leonel Malacrida

E-Mail Website
Guest Editor
1. Associate Professor, Department of Pathophysiology, Hospital de Clínicas, Facultad de Medicína, Universidad de la República-Uruguay, Montevideo- Uruguay
2. Advanced Bioimaging Unit, Institute Pasteur of Montevideo, Uruguay
Interests: biophotonics; FLIM; phasor plots; autofluorescence; FRET; environmental sensitive probes; deep tissue imaging.
Dr. Per Niklas Hedde

E-Mail Website
Guest Editor
Department of Pharmaceutical Sciences, University of California Irvine, Irvine, CA 92697, USA
Interests: biophysics; optics; image correlation spectroscopy; fluorescence lifetime and spectral detection; metabolic imaging; immune cell receptor dynamics.

Special Issue Information

Dear Colleagues,

Fluorescence imaging has a tremendous impact in life science. In particular, the advent of the novel combination of spectroscopy with the design of chemical and biological biosensors make it possible to interrogate in vivo cell processes that before needed to be done in vitro. The application of these novels biosensors changes the output form raw imagens to quantitative biology, where these methods have a strong impact in biophysical and biological systems. The quantitative information provided in fluorescence gives us detailed maps of cellular processes at unprecedented spatial and temporal resolution. Sensors – in terms of fluorescence species, can be broadly separated into two types: (i) biosensors, to study biological processes and (ii) instrument sensors, which can be used for influencing the imaging.

This Special Issue will cover state-of-the-art developments in biosensors and their applications for life science.

Dr. Suman Ranjit
Dr. Leonel Malacrida
Dr. Per Niklas Hedde
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. Sensors is an international peer-reviewed open access semimonthly 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 2600 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

  • Fluorescence
  • Biosensors
  • Image sensors
  • FLIM
  • Hyperspectral imaging

Published Papers (5 papers)

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Research

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18 pages, 5772 KiB  
Article
Generation and Characterization of Stable Redox-Reporter Mammalian Cell Lines of Biotechnological Relevance
by Karen Perelmuter, Inés Tiscornia, Marcelo A. Comini and Mariela Bollati-Fogolín
Sensors 2022, 22(4), 1324; https://doi.org/10.3390/s22041324 - 9 Feb 2022
Viewed by 1946
Abstract
Cellular functions such as DNA replication and protein translation are influenced by changes in the intracellular redox milieu. Exogenous (i.e., nutrients, deterioration of media components, xenobiotics) and endogenous factors (i.e., metabolism, growth) may alter the redox homeostasis of cells. Thus, monitoring redox changes [...] Read more.
Cellular functions such as DNA replication and protein translation are influenced by changes in the intracellular redox milieu. Exogenous (i.e., nutrients, deterioration of media components, xenobiotics) and endogenous factors (i.e., metabolism, growth) may alter the redox homeostasis of cells. Thus, monitoring redox changes in real time and in situ is deemed essential for optimizing the production of recombinant proteins. Recently, different redox-sensitive variants of green fluorescent proteins (e.g., rxYFP, roGFP2, and rxmRuby2) have been engineered and proved suitable to detect, in a non-invasive manner, perturbations in the pool of reduced and oxidized glutathione, the major low molecular mass thiol in mammals. In this study, we validate the use of cytosolic rxYFP on two cell lines widely used in biomanufacturing processes, namely, CHO-K1 cells expressing the human granulocyte macrophage colony-stimulating factor (hGM-CSF) and HEK-293. Flow cytometry was selected as the read-out technique for rxYFP signal given its high-throughput and statistical robustness. Growth kinetics and cellular metabolism (glucose consumption, lactate and ammonia production) of the redox reporter cells were comparable to those of the parental cell lines. The hGM-CSF production was not affected by the expression of the biosensor. The redox reporter cell lines showed a sensitive and reversible response to different redox stimuli (reducing and oxidant reagents). Under batch culture conditions, a significant and progressive oxidation of the biosensor occurred when CHO-K1-hGM-CSF cells entered the late-log phase. Medium replenishment restored, albeit partially, the intracellular redox homeostasis. Our study highlights the utility of genetically encoded redox biosensors to guide metabolic engineering or intervention strategies aimed at optimizing cell viability, growth, and productivity. Full article
(This article belongs to the Special Issue Sensors in Fluorescence Imaging)
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13 pages, 2394 KiB  
Article
Measuring Nanoscale Distances by Structured Illumination Microscopy and Image Cross-Correlation Spectroscopy (SIM-ICCS)
by Isotta Cainero, Elena Cerutti, Mario Faretta, Gaetano Ivan Dellino, Pier Giuseppe Pelicci, Alberto Diaspro and Luca Lanzanò
Sensors 2021, 21(6), 2010; https://doi.org/10.3390/s21062010 - 12 Mar 2021
Cited by 8 | Viewed by 3032
Abstract
Since the introduction of super-resolution microscopy, there has been growing interest in quantifying the nanoscale spatial distributions of fluorescent probes to better understand cellular processes and their interactions. One way to check if distributions are correlated or not is to perform colocalization analysis [...] Read more.
Since the introduction of super-resolution microscopy, there has been growing interest in quantifying the nanoscale spatial distributions of fluorescent probes to better understand cellular processes and their interactions. One way to check if distributions are correlated or not is to perform colocalization analysis of multi-color acquisitions. Among all the possible methods available to study and quantify the colocalization between multicolor images, there is image cross-correlation spectroscopy (ICCS). The main advantage of ICCS, in comparison with other co-localization techniques, is that it does not require pre-segmentation of the sample into single objects. Here we show that the combination of structured illumination microscopy (SIM) with ICCS (SIM-ICCS) is a simple approach to quantify colocalization and measure nanoscale distances from multi-color SIM images. We validate the SIM-ICCS analysis on SIM images of optical nanorulers, DNA-origami-based model samples containing fluorophores of different colors at a distance of 80 nm. The SIM-ICCS analysis is compared with an object-based analysis performed on the same samples. Finally, we show that SIM-ICCS can be used to quantify the nanoscale spatial distribution of functional nuclear sites in fixed cells. Full article
(This article belongs to the Special Issue Sensors in Fluorescence Imaging)
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14 pages, 5554 KiB  
Article
Hyperdimensional Imaging Contrast Using an Optical Fiber
by Jenu V. Chacko, Han Nim Lee, Wenxin Wu, Marisa S. Otegui and Kevin W. Eliceiri
Sensors 2021, 21(4), 1201; https://doi.org/10.3390/s21041201 - 9 Feb 2021
Cited by 2 | Viewed by 2202
Abstract
Fluorescence properties of a molecule can be used to study the structural and functional nature of biological processes. Physical properties, including fluorescence lifetime, emission spectrum, emission polarization, and others, help researchers probe a molecule, produce desired effects, and infer causes and consequences. Correlative [...] Read more.
Fluorescence properties of a molecule can be used to study the structural and functional nature of biological processes. Physical properties, including fluorescence lifetime, emission spectrum, emission polarization, and others, help researchers probe a molecule, produce desired effects, and infer causes and consequences. Correlative imaging techniques such as hyperdimensional imaging microscopy (HDIM) combine the physical properties and biochemical states of a fluorophore. Here we present a fiber-based imaging system that can generate hyper-dimensional contrast by combining multiple fluorescence properties into a single fluorescence lifetime decay curve. Fluorescence lifetime imaging microscopy (FLIM) with controlled excitation polarization and temporally dispersed emission can generate a spectrally coded, polarization-filtered lifetime distribution for a pixel. This HDIM scheme generates a better contrast between different molecules than that from individual techniques. This setup uses only a single detector and is simpler to implement, modular, cost-efficient, and adaptable to any existing FLIM microscope. We present higher contrast data from Arabidopsis thaliana epidermal cells based on intrinsic anthocyanin emission properties under multiphoton excitation. This work lays the foundation for an alternative hyperdimensional imaging system and demonstrates that contrast-based imaging is useful to study cellular heterogeneity in biological samples. Full article
(This article belongs to the Special Issue Sensors in Fluorescence Imaging)
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40 pages, 11977 KiB  
Article
Characterization of FIREFLY, an Imaging Spectrometer Designed for Remote Sensing of Solar Induced Fluorescence
by Ian Paynter, Bruce Cook, Lawrence Corp, Jyoteshwar Nagol and Joel McCorkel
Sensors 2020, 20(17), 4682; https://doi.org/10.3390/s20174682 - 19 Aug 2020
Cited by 7 | Viewed by 3155
Abstract
Solar induced fluorescence (SIF) is an ecological variable of interest to remote sensing retrievals, as it is directly related to vegetation composition and condition. FIREFLY (fluorescence imaging of red and far-red light yield) is a high performance spectrometer for estimating SIF. FIREFLY was [...] Read more.
Solar induced fluorescence (SIF) is an ecological variable of interest to remote sensing retrievals, as it is directly related to vegetation composition and condition. FIREFLY (fluorescence imaging of red and far-red light yield) is a high performance spectrometer for estimating SIF. FIREFLY was flown in conjunction with NASA Goddard’s lidar, hyperspectral, and thermal (G-LiHT) instrument package in 2017, as a technology demonstration for airborne retrievals of SIF. Attributes of FIREFLY relevant to SIF retrieval, including detector response and linearity; full-width at half maximum (FWHM); stray light; dark current; and shot noise were characterized with a combination of observations from Goddard’s laser for absolute measurement of radiance calibration facility; an integrating sphere; controlled acquisitions of known targets; in-flight acquisitions; and forward modelling. FWHM, stray light, and dark current were found to be of acceptable magnitude, and characterized to within acceptable limits for SIF retrieval. FIREFLY observations were found to represent oxygen absorption features, along with a large number of solar absorption features. Shot noise was acceptable for direct SIF retrievals at native resolution, but indirect SIF retrievals from absorption features would require spatial aggregation, or repeated observations of targets. Full article
(This article belongs to the Special Issue Sensors in Fluorescence Imaging)
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Review

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27 pages, 7165 KiB  
Review
Linear Combination Properties of the Phasor Space in Fluorescence Imaging
by Belén Torrado, Leonel Malacrida and Suman Ranjit
Sensors 2022, 22(3), 999; https://doi.org/10.3390/s22030999 - 27 Jan 2022
Cited by 13 | Viewed by 4515
Abstract
The phasor approach to fluorescence lifetime imaging, and more recently hyperspectral fluorescence imaging, has increased the use of these techniques, and improved the ease and intuitiveness of the data analysis. The fit-free nature of the phasor plots increases the speed of the analysis [...] Read more.
The phasor approach to fluorescence lifetime imaging, and more recently hyperspectral fluorescence imaging, has increased the use of these techniques, and improved the ease and intuitiveness of the data analysis. The fit-free nature of the phasor plots increases the speed of the analysis and reduces the dimensionality, optimization of data handling and storage. The reciprocity principle between the real and imaginary space—where the phasor and the pixel that the phasor originated from are linked and can be converted from one another—has helped the expansion of this method. The phasor coordinates calculated from a pixel, where multiple fluorescent species are present, depends on the phasor positions of those components. The relative positions are governed by the linear combination properties of the phasor space. According to this principle, the phasor position of a pixel with multiple components lies inside the polygon whose vertices are occupied by the phasor positions of these individual components and the distance between the image phasor to any of the vertices is inversely proportional to the fractional intensity contribution of that component to the total fluorescence from that image pixel. The higher the fractional intensity contribution of a vertex, the closer is the resultant phasor. The linear additivity in the phasor space can be exploited to obtain the fractional intensity contribution from multiple species and quantify their contribution. This review details the various mathematical models that can be used to obtain two/three/four components from phasor space with known phasor signatures and then how to obtain both the fractional intensities and phasor positions without any prior knowledge of either, assuming they are mono-exponential in nature. We note that other than for blind components, there are no restrictions on the type of the decay or their phasor positions for linear combinations to be valid—and they are applicable to complicated fluorescence lifetime decays from components with intensity decays described by multi-exponentials. Full article
(This article belongs to the Special Issue Sensors in Fluorescence Imaging)
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