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Applications of Fluorescence Microscopy in Molecular Biology
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Applications of Fluorescence Microscopy in Molecular Biology

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

Deadline for manuscript submissions: 30 June 2024 | Viewed by 6076

Special Issue Editor

Dr. Sang-Won Lee

E-Mail Website
Guest Editor
Center for Nano-Bio Measurement, Korea Research Institute of Standards and Science, 267 Gajeong-ro, Yuseong-gu, Daejeon 34113, Korea
Interests: optical microscopy; confocal fluorescence microscopy; two-photon excitation fluorescence microscopy; 3D deconvolution microscopy

Special Issue Information

Dear Colleagues,

As is well known, fluorescence microscopy is widely used to understand biological systems from nano-scale to micro-scale. Fluorescence microscopy has traditionally been an essential tool for monitoring cell physiology. Although the resolution of fluorescence microscopy is lower than that of electron microscopy, the recent development of fluorescence microscopy has made it possible to image live cells and in-vivo tissues in three dimensions. In addition, 3D-cell culture techniques (such as spheroids and organoids), optical clearing techniques, and new fluorescence dyes (such as quantum dots) have expanded the application scope of fluorescence microscopy to bioengineering, precision medicine, and high-contents-screening for new drug development. In this regard, sample preparation protocols and measurement protocols have also become important. In this context, this special issue aims to cover all areas, from the latest technologies in fluorescence microscopy to their applications. It welcomes original research, review articles, and short communications.

Dr. Sang-Won Lee
Guest Editor

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

  • single and two-photon fluorescence microscopy
  • super-resolution microscopy
  • 3D deconvolution microscopy
  • light-sheet microscopy
  • molecular imaging
  • organoids
  • brain imaging
  • drug screening
  • optical clearing
  • protocol

Published Papers (3 papers)

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Research

13 pages, 3034 KiB  
Article
Effects of Paclitaxel on Plasma Membrane Microviscosity and Lipid Composition in Cancer Cells
by Liubov Shimolina, Alexander Gulin, Alexandra Khlynova, Nadezhda Ignatova, Irina Druzhkova, Margarita Gubina, Elena Zagaynova, Marina K. Kuimova and Marina Shirmanova
Int. J. Mol. Sci. 2023, 24(15), 12186; https://doi.org/10.3390/ijms241512186 - 29 Jul 2023
Viewed by 1215
Abstract
The cell membrane is an important regulator for the cytotoxicity of chemotherapeutic agents. However, the biochemical and biophysical effects that occur in the membrane under the action of chemotherapy drugs are not fully described. In the present study, changes in the microviscosity of [...] Read more.
The cell membrane is an important regulator for the cytotoxicity of chemotherapeutic agents. However, the biochemical and biophysical effects that occur in the membrane under the action of chemotherapy drugs are not fully described. In the present study, changes in the microviscosity of membranes of living HeLa–Kyoto tumor cells were studied during chemotherapy with paclitaxel, a widely used antimicrotubule agent. To visualize the microviscosity of the membranes, fluorescence lifetime imaging microscopy (FLIM) with a BODIPY 2 fluorescent molecular rotor was used. The lipid profile of the membranes was assessed using time-of-flight secondary ion mass spectrometry ToF-SIMS. A significant, steady-state decrease in the microviscosity of membranes, both in cell monolayers and in tumor spheroids, was revealed after the treatment. Mass spectrometry showed an increase in the unsaturated fatty acid content in treated cell membranes, which may explain, at least partially, their low microviscosity. These results indicate the involvement of membrane microviscosity in the response of tumor cells to paclitaxel treatment. Full article
(This article belongs to the Special Issue Applications of Fluorescence Microscopy in Molecular Biology)
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14 pages, 2870 KiB  
Article
Extended Depth of Focus Two-Photon Light-Sheet Microscopy for In Vivo Fluorescence Imaging of Large Multicellular Organisms at Cellular Resolution
by Takashi Saitou and Takeshi Imamura
Int. J. Mol. Sci. 2023, 24(12), 10186; https://doi.org/10.3390/ijms241210186 - 15 Jun 2023
Viewed by 1177
Abstract
Two-photon excitation in light-sheet microscopy advances applications to live imaging of multicellular organisms. In a previous study, we developed a two-photon Bessel beam light-sheet microscope with a nearly 1-mm field of view and less than 4-μm axial resolution, using a low magnification (10×), [...] Read more.
Two-photon excitation in light-sheet microscopy advances applications to live imaging of multicellular organisms. In a previous study, we developed a two-photon Bessel beam light-sheet microscope with a nearly 1-mm field of view and less than 4-μm axial resolution, using a low magnification (10×), middle numerical aperture (NA 0.5) detection objective. In this study, we aimed to construct a light-sheet microscope with higher resolution imaging while maintaining the large field of view, using low magnification (16×) with a high NA 0.8 objective. To address potential illumination and detection mismatch, we investigated the use of a depth of focus (DOF) extension method. Specifically, we used a stair-step device composed of five-layer annular zones that extended DOF two-fold, enough to cover the light-sheet thickness. Resolution measurements using fluorescent beads showed that the reduction in resolutions was small. We then applied this system to in vivo imaging of medaka fish and found that image quality degradation at the distal site of the beam injection could be compensated. This demonstrates that the extended DOF system combined with wide-field two-photon light-sheet microscopy offers a simple and easy setup for live imaging application of large multicellular organism specimens with sub-cellular resolution. Full article
(This article belongs to the Special Issue Applications of Fluorescence Microscopy in Molecular Biology)
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17 pages, 12025 KiB  
Article
Characterizing and Quenching Autofluorescence in Fixed Mouse Adrenal Cortex Tissue
by Nawar Sakr, Olga Glazova, Liudmila Shevkova, Nikita Onyanov, Samira Kaziakhmedova, Alena Shilova, Maria V. Vorontsova and Pavel Volchkov
Int. J. Mol. Sci. 2023, 24(4), 3432; https://doi.org/10.3390/ijms24043432 - 08 Feb 2023
Cited by 3 | Viewed by 3086
Abstract
Tissue autofluorescence of fixed tissue sections is a major concern of fluorescence microscopy. The adrenal cortex emits intense intrinsic fluorescence that interferes with signals from fluorescent labels, resulting in poor-quality images and complicating data analysis. We used confocal scanning laser microscopy imaging and [...] Read more.
Tissue autofluorescence of fixed tissue sections is a major concern of fluorescence microscopy. The adrenal cortex emits intense intrinsic fluorescence that interferes with signals from fluorescent labels, resulting in poor-quality images and complicating data analysis. We used confocal scanning laser microscopy imaging and lambda scanning to characterize the mouse adrenal cortex autofluorescence. We evaluated the efficacy of tissue treatment methods in reducing the intensity of the observed autofluorescence, such as trypan blue, copper sulfate, ammonia/ethanol, Sudan Black B, TrueVIEWTM Autofluorescence Quenching Kit, MaxBlockTM Autofluorescence Reducing Reagent Kit, and TrueBlackTM Lipofuscin Autofluorescence Quencher. Quantitative analysis demonstrated autofluorescence reduction by 12–95%, depending on the tissue treatment method and excitation wavelength. TrueBlackTM Lipofuscin Autofluorescence Quencher and MaxBlockTM Autofluorescence Reducing Reagent Kit were the most effective treatments, reducing the autofluorescence intensity by 89–93% and 90–95%, respectively. The treatment with TrueBlackTM Lipofuscin Autofluorescence Quencher preserved the specific fluorescence signals and tissue integrity, allowing reliable detection of fluorescent labels in the adrenal cortex tissue. This study demonstrates a feasible, easy-to-perform, and cost-effective method to quench tissue autofluorescence and improve the signal-to-noise ratio in adrenal tissue sections for fluorescence microscopy. Full article
(This article belongs to the Special Issue Applications of Fluorescence Microscopy in Molecular Biology)
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