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Application of Fluorescence Imaging and Super-Resolution Imaging in Molecular Biology
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Application of Fluorescence Imaging and Super-Resolution Imaging in Molecular Biology

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

Deadline for manuscript submissions: 20 March 2025 | Viewed by 3714

Special Issue Editor

Dr. Keigo Kohara

E-Mail Website
Guest Editor
KMU Biobank Center, Institute of Biomedical Science, Kansai Medical University, 573-1010 Hirakata, Japan
Interests: super-resolution imaging; molecular biology; neuroscience; hippocampus; new CA2 region; new technologies; BDNF

Special Issue Information

Dear Colleagues,

Recent inventions of super-resolution microscopies have induced a revolution in molecular biology and enabled us to see various biological molecules, organelles, cells, and disease events at the nanoscale. In addition, evolutions of fluorescence imaging technologies have also contributed to progress in various areas of molecular biology. International Journal of Molecular Sciences will publish a Special Issue in 2023 to highlight studies in molecular biology that use fluorescence imaging and super-resolution microscopy.

We therefore invite reviews and research articles on the application of fluorescence imaging in molecular biology and super-resolution imaging in molecular biology.

We will collect a series of articles that include fluorescence imaging and super-resolution imaging in molecular biology, molecular disease biology, and methodologies of fluorescence and super-resolution imaging.

The related topics include, but are not limited to:

  • Fluorescence imaging of biomolecules or organelles;
  • Fluorescence imaging of the morphologies of cells;
  • Fluorescence imaging in iPS cells or organoids;
  • Fluorescence imaging in animal disease models;
  • Methodologies of fluorescence imaging for molecular biology;
  • Super-resolution imaging of biomolecules or organelles;
  • Super-resolution imaging of the morphologies of cells;
  • Super-resolution imaging in animal disease models;
  • Super-resolution imaging in iPS cells or organoids;
  • Methodologies of super-resolution microscopies for molecular biology.

Dr. Keigo Kohara
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

  • fluorescence imaging
  • super-resolution imaging
  • molecular biology
  • disease
  • biomolecules
  • organelles
  • morphologies of cells
  • iPS cells
  • organoids
  • molecular biology

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Further information on MDPI's Special Issue polices can be found here.

Published Papers (3 papers)

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Research

12 pages, 5537 KiB  
Article
Accompanying Hemoglobin Polymerization in Red Blood Cells in Patients with Sickle Cell Disease Using Fluorescence Lifetime Imaging
by Fernanda Aparecida Borges da Silva, João Batista Florindo, Amilcar Castro de Mattos, Fernando Ferreira Costa, Irene Lorand-Metze and Konradin Metze
Int. J. Mol. Sci. 2024, 25(22), 12290; https://doi.org/10.3390/ijms252212290 - 15 Nov 2024
Viewed by 493
Abstract
In recent studies, it has been shown that fluorescence lifetime imaging (FLIM) may reveal intracellular structural details in unstained cytological preparations that are not revealed by standard staining procedures. The aim of our investigation was to examine whether FLIM images could reveal areas [...] Read more.
In recent studies, it has been shown that fluorescence lifetime imaging (FLIM) may reveal intracellular structural details in unstained cytological preparations that are not revealed by standard staining procedures. The aim of our investigation was to examine whether FLIM images could reveal areas suggestive of polymerization in red blood cells (RBCs) of sickle cell disease (SCD) patients. We examined label-free blood films using auto-fluorescence FLIM images of 45 SCD patients and compared the results with those of 27 control persons without hematological disease. All control RBCs revealed homogeneous cytoplasm without any foci. Rounded non-sickled RBCs in SCD showed between zero and three small intensively fluorescent dots with higher lifetime values. In sickled RBCs, we found additionally larger irregularly shaped intensively fluorescent areas with increased FLIM values. These areas were interpreted as equivalent to polymerized hemoglobin. The rounded, non-sickled RBCs of SCD patients with homogeneous cytoplasm were not different from those of the erythrocytes of control patients in light microscopy. Yet, variables from the local binary pattern-transformed matrix of the FLIM values per pixel showed significant differences between non-sickled RBCs and those of control cells. In a linear discriminant analysis, using local binary pattern-transformed texture features (mean and entropy) of the erythrocyte cytoplasm of normal appearing cells, the final model could distinguish between SCD patients and control persons with an accuracy of 84.7% of the patients. When the classification was based on the examination of a single rounded erythrocyte, an accuracy of 68.5% was achieved. Employing the Linear Discriminant Analysis classifier method for machine learning, the accuracy was 68.1%. We believe that our study shows that FLIM is able to disclose the topography of the intracellular polymerization process of hemoglobin in sickle cell disease and that the images are compatible with the theory of the two-step nucleation. Furthermore, we think that the presented technique may be an interesting tool for the investigation of therapeutic inhibition of polymerization. Full article
(This article belongs to the Special Issue Application of Fluorescence Imaging and Super-Resolution Imaging in Molecular Biology)
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24 pages, 7912 KiB  
Article
Altered Endoplasmic Reticulum Integrity and Organelle Interactions in Living Cells Expressing INF2 Variants
by Quynh Thuy Huong Tran, Naoyuki Kondo, Hiroko Ueda, Yoshiyuki Matsuo and Hiroyasu Tsukaguchi
Int. J. Mol. Sci. 2024, 25(18), 9783; https://doi.org/10.3390/ijms25189783 - 10 Sep 2024
Viewed by 886
Abstract
The cytoskeleton mediates fundamental cellular processes by organizing inter-organelle interactions. Pathogenic variants of inverted formin 2 (INF2) CAAX isoform, an actin assembly factor that is predominantly expressed in the endoplasmic reticulum (ER), are linked to focal segmental glomerulosclerosis (FSGS) and Charcot–Marie–Tooth (CMT) neuropathy. [...] Read more.
The cytoskeleton mediates fundamental cellular processes by organizing inter-organelle interactions. Pathogenic variants of inverted formin 2 (INF2) CAAX isoform, an actin assembly factor that is predominantly expressed in the endoplasmic reticulum (ER), are linked to focal segmental glomerulosclerosis (FSGS) and Charcot–Marie–Tooth (CMT) neuropathy. To investigate how pathogenic INF2 variants alter ER integrity, we used high-resolution live imaging of HeLa cells. Cells expressing wild-type (WT) INF2 showed a predominant tubular ER with perinuclear clustering. Cells expressing INF2 FSGS variants that cause mild and intermediate disease induced more sheet-like ER, a pattern similar to that seen for cells expressing WT-INF2 that were treated with actin and microtubule (MT) inhibitors. Dual CMT-FSGS INF2 variants led to more severe ER dysmorphism, with a diffuse, fragmented ER and coarse INF2 aggregates. Proper organization of both F-actin and MT was needed to modulate the tubule vs. sheet conformation balance, while MT arrays regulated spatial expansion of tubular ER in the cell periphery. Pathogenic INF2 variants also induced mitochondria fragmentation and dysregulated mitochondria distribution. Such mitochondrial abnormalities were more prominent for cells expressing CMT-FSGS compared to those with FSGS variants, indicating that the severity of the dysfunction is linked to the degree of cytoskeletal disorganization. Our observations suggest that pathogenic INF2 variants disrupt ER continuity by altering interactions between the ER and the cytoskeleton that in turn impairs inter-organelle communication, especially at ER–mitochondria contact sites. ER continuity defects may be a common disease mechanism involved in both peripheral neuropathy and glomerulopathy. Full article
(This article belongs to the Special Issue Application of Fluorescence Imaging and Super-Resolution Imaging in Molecular Biology)
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21 pages, 20175 KiB  
Article
From Cell Populations to Molecular Complexes: Multiplexed Multimodal Microscopy to Explore p53-53BP1 Molecular Interaction
by Simone Pelicci, Laura Furia, Pier Giuseppe Pelicci and Mario Faretta
Int. J. Mol. Sci. 2024, 25(9), 4672; https://doi.org/10.3390/ijms25094672 - 25 Apr 2024
Viewed by 1328
Abstract
Surpassing the diffraction barrier revolutionized modern fluorescence microscopy. However, intrinsic limitations in statistical sampling, the number of simultaneously analyzable channels, hardware requirements, and sample preparation procedures still represent an obstacle to its widespread diffusion in applicative biomedical research. Here, we present a novel [...] Read more.
Surpassing the diffraction barrier revolutionized modern fluorescence microscopy. However, intrinsic limitations in statistical sampling, the number of simultaneously analyzable channels, hardware requirements, and sample preparation procedures still represent an obstacle to its widespread diffusion in applicative biomedical research. Here, we present a novel pipeline based on automated multimodal microscopy and super-resolution techniques employing easily available materials and instruments and completed with open-source image-analysis software developed in our laboratory. The results show the potential impact of single-molecule localization microscopy (SMLM) on the study of biomolecules’ interactions and the localization of macromolecular complexes. As a demonstrative application, we explored the basis of p53-53BP1 interactions, showing the formation of a putative macromolecular complex between the two proteins and the basal transcription machinery in situ, thus providing visual proof of the direct role of 53BP1 in sustaining p53 transactivation function. Moreover, high-content SMLM provided evidence of the presence of a 53BP1 complex on the cell cytoskeleton and in the mitochondrial space, thus suggesting the existence of novel alternative 53BP1 functions to support p53 activity. Full article
(This article belongs to the Special Issue Application of Fluorescence Imaging and Super-Resolution Imaging in Molecular Biology)
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