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New Trends in Fluorescence Correlation Spectroscopy and Its Application Methods
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New Trends in Fluorescence Correlation Spectroscopy and Its Application Methods

A special issue of Applied Sciences (ISSN 2076-3417). This special issue belongs to the section "Applied Biosciences and Bioengineering".

Deadline for manuscript submissions: closed (21 January 2022) | Viewed by 12447

Special Issue Editors

Dr. Akira Kitamura

E-Mail Website
Guest Editor
Faculty of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan
Interests: protein aggregation; protein folding/misfolding; molecular chaperone; proteostasis; neurodegenerative disease; fluorescence spectroscopy; fluorescence correlation spectroscopy; fluorescence lifetime
Dr. Akira Sasaki

E-Mail Website
Guest Editor
Bioanalytical Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan
Interests: biological standardization; reference materials; nucleic acids; fluorescence correlation spectroscopy; image correlation spectroscopy; STED nanoscopy

Special Issue Information

Dear Colleagues,

Fluorescence correlation spectroscopy (FCS) has significantly contributed to the understanding of molecular dynamics in a lot of biological systems such as diffusion, interaction, oligomerization/aggregation of auto-fluorescent or fluorescent dye-labeled biomolecules. Many researchers believe that FCS does not include any limitations for the application because the limitation is just a restriction of the idea. The related methods and applications of FCS have been widely established after the establishment of a robust basic principle of FCS.

This special issue focuses on the vast range of FCS and its advanced methods not only in biological applications but also as basic improvements of FCS or correlation spectroscopy. Improvement of equipment and optical system, the establishment of statistical analysis methods, analysis methods using computational simulation, and so on, are required in typical advances of FCS. High-throughput screening or standardization and calibration strategies using FCS are also important for the biomedical and biophysical fields.

FCS has been widely involved in a variety of research fields such as molecular and cellular biology, biochemistry, biophysics, pharmacology and biomedicine, and others. The latest technological developments and biological applications using FCS and its related methods will be shared through this special issue. We invite researchers and investigators to contribute their original research or review articles to this special issue.

Dr. Akira Kitamura
Dr. Akira Sasaki
Guest Editors

Manuscript Submission Information

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Keywords

  • Fluorescence correlation spectroscopy
  • Image correlation spectroscopy
  • Physical chemistry of fluorescence
  • Biophysics
  • Biochemistry
  • Molecular and cellular biology
  • Diffusion of molecules
  • Interaction of molecules
  • Oligomerization/aggregation of molecules

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Published Papers (4 papers)

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Research

8 pages, 1644 KiB  
Article
Monitoring Molecular Properties of a Fluorescence Light-Up Aptamer Using Fluorescence Cross-Correlation Spectroscopy
by Yuichi Furuhata and Akira Sasaki
Appl. Sci. 2022, 12(4), 2002; https://doi.org/10.3390/app12042002 - 14 Feb 2022
Cited by 1 | Viewed by 2801
Abstract
Fluorescence light-up aptamers (FLAPs) are tools for RNA imaging, wherein the RNA of interest is appended with a FLAP sequence that can bind to a corresponding small-molecule fluorogen and enhance its fluorescence. The fluorescence properties of FLAPs have mostly been analyzed in bulk [...] Read more.
Fluorescence light-up aptamers (FLAPs) are tools for RNA imaging, wherein the RNA of interest is appended with a FLAP sequence that can bind to a corresponding small-molecule fluorogen and enhance its fluorescence. The fluorescence properties of FLAPs have mostly been analyzed in bulk and described as the average of a large number of RNA–fluorogen complexes. In this study, we evaluated the feasibility of fluorescence correlation spectroscopy (FCS)- and fluorescence cross-correlation spectroscopy (FCCS)-based quantifications of FLAPs in a solution using Broccoli, a common FLAP, and its corresponding fluorogen, DFHBI-1T. We investigated the folding efficiency, photostability, and photophysical properties of the Broccoli–DFHBI-1T complex using their FCS/FCCS characteristics. With FCS, we observed that the fluorescence was affected by the affinity between Broccoli and DFHBI-1T and the folding (maturation) state of Broccoli RNA. Moreover, the FCCS measurement of ATTO647N-labeled Broccoli and its complex with DFHBI-1T revealed the proportion of the mature Broccoli–DFHBI-1T complex. The current FCS/FCCS-based study of Broccoli–DFHBI-1T provides a model for analyzing FLAPs and their fluorogen pairs at the single-molecule level. Full article
(This article belongs to the Special Issue New Trends in Fluorescence Correlation Spectroscopy and Its Application Methods)
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10 pages, 6292 KiB  
Article
Interaction between Spike Protein of SARS-CoV-2 and Human Virus Receptor ACE2 Using Two-Color Fluorescence Cross-Correlation Spectroscopy
by Ai Fujimoto, Yidan Lyu, Masataka Kinjo and Akira Kitamura
Appl. Sci. 2021, 11(22), 10697; https://doi.org/10.3390/app112210697 - 12 Nov 2021
Cited by 3 | Viewed by 3919
Abstract
Infection with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the cause of coronavirus disease 2019 (COVID-19), is initiated by the interaction between a receptor protein, angiotensin-converting enzyme type 2 (ACE2) on the cell surface, and the viral spike (S) protein. This interaction is similar [...] Read more.
Infection with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the cause of coronavirus disease 2019 (COVID-19), is initiated by the interaction between a receptor protein, angiotensin-converting enzyme type 2 (ACE2) on the cell surface, and the viral spike (S) protein. This interaction is similar to the mechanism in SARS-CoV, a close relative of SARS-CoV-2, which was identified in 2003. Drugs and antibodies that inhibit the interaction between ACE2 and S proteins could be key therapeutic methods for preventing viral infection and replication in COVID-19. Here, we demonstrate the interaction between human ACE2 and a fragment of the S protein (S1 subunit) derived from SARS-CoV-2 and SARS-CoV using two-color fluorescence cross-correlation spectroscopy (FCCS), which can detect the interaction of fluorescently labeled proteins. The S1 subunit of SARS-CoV-2 interacted in solution with soluble ACE2, which lacks a transmembrane region, more strongly than that of SARS-CoV. Furthermore, one-to-one stoichiometry of the two proteins during the interaction was indicated. Thus, we propose that this FCCS-based interaction detection system can be used to analyze the interaction strengths of various mutants of the S1 subunit that have evolved during the worldwide pandemic, and also offers the opportunity to screen and evaluate the performance of drugs and antibodies that inhibit the interaction. Full article
(This article belongs to the Special Issue New Trends in Fluorescence Correlation Spectroscopy and Its Application Methods)
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10 pages, 1488 KiB  
Article
Fluorescence Correlation Spectroscopy Measurement Based on Fiber Optics for Biological Materials
by Johtaro Yamamoto and Akira Sasaki
Appl. Sci. 2021, 11(15), 6744; https://doi.org/10.3390/app11156744 - 22 Jul 2021
Cited by 1 | Viewed by 2766
Abstract
A robust fluorescence correlation spectroscopy system called fiber-optic based fluorescence correlation spectroscopy (FB-FCS) was developed; this system enables the measurement of diffusion dynamics and concentration of fluorescent molecules based on the principle of fluorescence correlation spectroscopy without any mechanical adjustment of the experimental [...] Read more.
A robust fluorescence correlation spectroscopy system called fiber-optic based fluorescence correlation spectroscopy (FB-FCS) was developed; this system enables the measurement of diffusion dynamics and concentration of fluorescent molecules based on the principle of fluorescence correlation spectroscopy without any mechanical adjustment of the experimental setup. The system consisted of fiber optics and a water-immersion objective lens. The hydrodynamic diameters and concentrations of organic fluorescent dyes and fluorescently labeled proteins were successfully measured. Because of the fiber-optic-based setup, the FB-FCS system is compact and inexpensive. We expect FB-FCS to be suitable for use in laboratories, medical diagnosis, and environmental measurements. Full article
(This article belongs to the Special Issue New Trends in Fluorescence Correlation Spectroscopy and Its Application Methods)
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10 pages, 1404 KiB  
Article
Measurement of the Concentration and the Brightness for Samples Containing Multiple Molecules with Different Brightness Using Fluorescence Correlation Spectroscopy
by Johtaro Yamamoto and Akira Sasaki
Appl. Sci. 2021, 11(13), 5840; https://doi.org/10.3390/app11135840 - 23 Jun 2021
Cited by 1 | Viewed by 1964
Abstract
In this study, the concentration and brightness measured by fluorescence correlation spectroscopy (FCS) in samples containing multiple species with different brightness levels was demonstrated. FCS measurements of such samples are generally difficult. However, the calculation we introduced here can provide the measurement results [...] Read more.
In this study, the concentration and brightness measured by fluorescence correlation spectroscopy (FCS) in samples containing multiple species with different brightness levels was demonstrated. FCS measurements of such samples are generally difficult. However, the calculation we introduced here can provide the measurement results of the FCS. The effectiveness of the calculation was investigated based on simulations and experiments in the case of a mixture of fluorescent beads with known brightness and other fluorescent beads with unknown brightness. The results show that the concentration of the known brightness agrees well with the expected values. The obtained concentration and brightness of the species with unknown brightness is possible, and it worked well in the simulation; however, the accuracy for the species was lower than that of the species with known brightness. As a result, the calculation is useful in measuring the concentration of species with known brightness in samples containing undesired bright species, such as aggregation. The calculation for the species with unknown brightness may also be useful if good protocols or instruments are established in the future. Full article
(This article belongs to the Special Issue New Trends in Fluorescence Correlation Spectroscopy and Its Application Methods)
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