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Cells
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Advanced Technology for Cellular Imaging
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Advanced Technology for Cellular Imaging

A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Cell Methods".

Deadline for manuscript submissions: 31 January 2025 | Viewed by 11354

Special Issue Editors

Dr. Alessandro Grinzato

E-Mail Website
Guest Editor
European Synchrotron Radiation Facility, 71 Avenue des Martyrs, 38000 Grenoble, France
Interests: structural biology; electron microscopy; protein–protein interaction; protein flexibility
Dr. Vittoria Raimondi

E-Mail Website
Guest Editor
Department of Surgery, Oncology and Gastroenterology–DiSCOG, University of Padova, Padova, Italy
Interests: cancer metabolism; redox homeostasis; leukemia
Dr. J. Bernard Heymann

E-Mail Website
Guest Editor
1. Laboratory for Structural Biology Research, NIAMS, NIH, Bethesda, MD 20892, USA
2. National Cryo-EM Program, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, MD 21701, USA
Interests: cryo-electron microscopy (or cryoeEM); cryo-electron tomography; structural biology; single-particle analysis; micrograph processing software; cellular segmentation; virology

Special Issue Information

Dear Colleagues,

Understanding the correlation between cellular organization and function is one of the major goals of biology. In this context, imaging techniques allow for cell investigation at the subcellular to the macromolecule level. Although confocal and electron microscopy have become commonly used techniques for cellular studies, new technological advances keep appearing, allowing us to obtain more and more information than before. Light microscopy can overcome the diffraction limit, and electron tomography can provide high-resolution protein structures. Moreover, multiparameter fluorescence imaging allows for concomitantly analyzing multiple biomarkers in the same sample by exploiting the whole spectrum. Finally, the possibility of performing electron and fluorescent microscopy on the same sample allows the researcher to identify labeled cell portions before their high-resolution characterization through EM. In parallel, computational methods, driven by the recent implementation of artificial intelligence, have become essential tools in improving and developing new cell imaging techniques. 

This Special Issue aims to examine the most recent advances in cell biology imaging techniques and discuss the future direction these techniques will take. Therefore, we welcome Original Research and Review articles covering the latest and current findings, methods, and applications in the wide field of cell imaging, with a specific interest in innovative approaches driven by recent technological advances. 

Dr. Alessandro Grinzato
Dr. Vittoria Raimondi
Dr. J. Bernard Heymann
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. Cells 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 2700 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

  • electron tomography
  • fluorescence microscopy
  • live cell imaging
  • multimodal imaging
  • super resolution microscopy
  • cell–cell interaction

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Published Papers (7 papers)

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Research

12 pages, 4118 KiB  
Article
Su(H) Modulates Enhancer Transcriptional Bursting in Prelude to Gastrulation
by Kelli D. Fenelon, Priyanshi Borad, Biraaj Rout, Parisa Boodaghi Malidarreh, Mohammad Sadegh Nasr, Jacob M. Luber and Theodora Koromila
Cells 2024, 13(21), 1759; https://doi.org/10.3390/cells13211759 - 24 Oct 2024
Viewed by 859
Abstract
Transcriptional regulation, orchestrated by the interplay between transcription factors (TFs) and enhancers, governs gene expression dynamics crucial for cellular processes. While gross qualitative fluctuations in transcription factor-dependent gene expression patterning have a long history of characterization, the roles of these factors in the [...] Read more.
Transcriptional regulation, orchestrated by the interplay between transcription factors (TFs) and enhancers, governs gene expression dynamics crucial for cellular processes. While gross qualitative fluctuations in transcription factor-dependent gene expression patterning have a long history of characterization, the roles of these factors in the nuclei retaining expression in the presence or absence of these factors are now observable using modern techniques. Our study investigates the impact of Suppressor of Hairless (Su(H)), a broadly expressed transcription factor, on enhancer-driven transcriptional modulation using Drosophila early embryos as a model system. Building upon previous findings, we employ super-resolution microscopy to dissect Su(H)’s influence on sog-Distal (sogD) enhancer activity specifically in nuclei with preserved sogD-driven expression in the absence of Su(H) binding. We demonstrate that Su(H) occupancy perturbations alter expression levels and bursting dynamics. Notably, Su(H) absence during embryonic development exhibits region-specific effects, inhibiting expression dorsally and stabilizing expression ventrally, implying a nuanced role in enhancer regulation. Our findings shed light on the intricate mechanisms that govern transcriptional dynamics and suggest a critical patterning role for Notch/Hairless signaling in sog expression as embryos transition to gastrulation. Full article
(This article belongs to the Special Issue Advanced Technology for Cellular Imaging)
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12 pages, 9826 KiB  
Article
Unveiling a Surgical Revolution: The Use of Conventional Histology versus Ex Vivo Fusion Confocal Microscopy in Breast Cancer Surgery
by Daniel Humaran, Javiera Pérez-Anker, Pedro L. Fernández, Lidia Blay, Iciar Pascual, Eva Castellà, Laia Pérez, Susana Puig, Josep Malvehy and Joan F. Julián
Cells 2024, 13(20), 1692; https://doi.org/10.3390/cells13201692 - 12 Oct 2024
Viewed by 921
Abstract
Ex vivo fusion confocal microscopy (EVFCM) enables the rapid examination of breast tissue and has the potential to reduce the surgical margins and the necessity for further surgeries. Traditional methods, such as frozen section analysis, are limited by the distortion of tissue and [...] Read more.
Ex vivo fusion confocal microscopy (EVFCM) enables the rapid examination of breast tissue and has the potential to reduce the surgical margins and the necessity for further surgeries. Traditional methods, such as frozen section analysis, are limited by the distortion of tissue and artefacts, leading to false negatives and the need for additional surgeries. This study on observational diagnostic accuracy evaluated the ability of EVFCM to detect breast cancer. A total of 36 breast tissue samples, comprising 20 non-neoplastic and 16 neoplastic cases, were analysed using EVFCM and compared to the results obtained from routine histopathology. A Mohs surgeon experienced in EVFCM (evaluator A) and two breast pathologists unfamiliar with EVFCM (evaluators B and C) performed blinded analyses. EVFCM showed high concordance with the histopathology and the detection of neoplasia, with significant kappa values (p < 0.001). Evaluator A achieved 100% sensitivity and specificity. Evaluators B and C achieved a sensitivity of >87%, a specificity of >94%, positive predictive values of >95%, and negative predictive values of 81% and 94%, respectively. EVFCM therefore offers a promising technique for the assessment of margins in breast-conserving surgery. Its widespread adoption could significantly reduce re-excisions, lower healthcare costs, and improve cosmetic and oncological outcomes. Full article
(This article belongs to the Special Issue Advanced Technology for Cellular Imaging)
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17 pages, 3344 KiB  
Article
Cell-Sonar, a Novel Method for Intracellular Tracking of Secretory Pathways
by Sabrina Brockmöller, Thomas Seeger, Franz Worek and Simone Rothmiller
Cells 2024, 13(17), 1449; https://doi.org/10.3390/cells13171449 - 29 Aug 2024
Viewed by 865
Abstract
Background: Intracellular tracking is commonly used in trafficking research. Until today, the respective techniques have remained complex, and complicated, mostly transgenic target protein changes are necessary, often requiring expensive equipment and expert knowledge. Methods: We present a novel method, which we term “cell-sonar”, [...] Read more.
Background: Intracellular tracking is commonly used in trafficking research. Until today, the respective techniques have remained complex, and complicated, mostly transgenic target protein changes are necessary, often requiring expensive equipment and expert knowledge. Methods: We present a novel method, which we term “cell-sonar”, that enables the user to track expression changes of specific protein markers that serve as points of interaction. Our study provides comparable analyses of expression changes of these marker proteins by in-cell Western analyses in two otherwise isogenic cell lines that only differ in the overexpression of the tracked target protein. Using the overexpressed human adult muscle-type nicotinic acetylcholine receptor as an example, we demonstrate that cell-sonar can cover multiple intracellular compartments such as the endoplasmic reticulum, the pathway between it and the Golgi apparatus, and the endocytic pathway. Results: We provide evidence for receptor maturation in the Golgi and storage in recycling endosomes, rather than the fate of increased insertion into the plasma membrane. Additionally, we demonstrate with the implementation of nicotine that the receptor’s destiny is exasperated up to secondary degradation. Conclusions: Cell-sonar is an affordable, easy-to-implement, and cheap method that can be adapted to a broad variety of proteins and cellular pathways of interest to researchers. Full article
(This article belongs to the Special Issue Advanced Technology for Cellular Imaging)
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12 pages, 1818 KiB  
Article
Precise Serial Microregistration Enables Quantitative Microscopy Imaging Tracking of Human Skin Cells In Vivo
by Yunxian Tian, Zhenguo Wu, Harvey Lui, Jianhua Zhao, Sunil Kalia, InSeok Seo, Hao Ou-Yang and Haishan Zeng
Cells 2024, 13(13), 1158; https://doi.org/10.3390/cells13131158 - 7 Jul 2024
Viewed by 1052
Abstract
We developed an automated microregistration method that enables repeated in vivo skin microscopy imaging of the same tissue microlocation and specific cells over a long period of days and weeks with unprecedented precision. Applying this method in conjunction with an in vivo multimodality [...] Read more.
We developed an automated microregistration method that enables repeated in vivo skin microscopy imaging of the same tissue microlocation and specific cells over a long period of days and weeks with unprecedented precision. Applying this method in conjunction with an in vivo multimodality multiphoton microscope, the behavior of human skin cells such as cell proliferation, melanin upward migration, blood flow dynamics, and epidermal thickness adaptation can be recorded over time, facilitating quantitative cellular dynamics analysis. We demonstrated the usefulness of this method in a skin biology study by successfully monitoring skin cellular responses for a period of two weeks following an acute exposure to ultraviolet light. Full article
(This article belongs to the Special Issue Advanced Technology for Cellular Imaging)
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16 pages, 17359 KiB  
Communication
Use of FRET-Sensor ‘Mermaid’ to Detect Subtle Changes in Membrane Potential of Primary Mouse PASMCs
by Ruth C. Dartsch, Simone Kraut, Tim Mayer, Andreas Gabel, Alexander Dietrich, Norbert Weissmann, Beate Fuchs and Fenja Knoepp
Cells 2024, 13(12), 1070; https://doi.org/10.3390/cells13121070 - 20 Jun 2024
Viewed by 3640
Abstract
Subtle changes in the membrane potential of pulmonary arterial smooth muscle cells (PASMCs) are pivotal for controlling pulmonary vascular tone, e.g., for initiating Hypoxic Pulmonary Vasoconstriction, a vital mechanism of the pulmonary circulation. In our study, we evaluated the ability of the fluorescence [...] Read more.
Subtle changes in the membrane potential of pulmonary arterial smooth muscle cells (PASMCs) are pivotal for controlling pulmonary vascular tone, e.g., for initiating Hypoxic Pulmonary Vasoconstriction, a vital mechanism of the pulmonary circulation. In our study, we evaluated the ability of the fluorescence resonance energy transfer (FRET)-based voltage-sensor Mermaid to detect such subtle changes in membrane potential. Mouse PASMCs were isolated and transduced with Mermaid-encoding lentiviral vectors before the acceptor/donor emission ratio was assessed via live cell FRET-imaging. Mermaid’s sensitivity was tested by applying specific potassium chloride (KCl) concentrations. These KCl concentrations were previously validated by patch clamp recordings to induce depolarization with predefined amplitudes that physiologically occur in PASMCs. Mermaid’s emission ratio dose-dependently increased upon depolarization with KCl. However, Mermaid formed unspecific intracellular aggregates, which limited the usefulness of this voltage sensor. When analyzing the membrane rim only to circumvent these unspecific signals, Mermaid was not suitable to resolve subtle changes in the membrane potential of ≤10 mV. In summary, we found Mermaid to be a suitable alternative for reliably detecting qualitative membrane voltage changes of more than 10 mV in primary mouse PASMCs. However, one should be aware of the limitations associated with this voltage sensor. Full article
(This article belongs to the Special Issue Advanced Technology for Cellular Imaging)
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Figure 1

19 pages, 20483 KiB  
Article
Subcellular Feature-Based Classification of α and β Cells Using Soft X-ray Tomography
by Aneesh Deshmukh, Kevin Chang, Janielle Cuala, Bieke Vanslembrouck, Senta Georgia, Valentina Loconte and Kate L. White
Cells 2024, 13(10), 869; https://doi.org/10.3390/cells13100869 - 18 May 2024
Viewed by 1761
Abstract
The dysfunction of α and β cells in pancreatic islets can lead to diabetes. Many questions remain on the subcellular organization of islet cells during the progression of disease. Existing three-dimensional cellular mapping approaches face challenges such as time-intensive sample sectioning and subjective [...] Read more.
The dysfunction of α and β cells in pancreatic islets can lead to diabetes. Many questions remain on the subcellular organization of islet cells during the progression of disease. Existing three-dimensional cellular mapping approaches face challenges such as time-intensive sample sectioning and subjective cellular identification. To address these challenges, we have developed a subcellular feature-based classification approach, which allows us to identify α and β cells and quantify their subcellular structural characteristics using soft X-ray tomography (SXT). We observed significant differences in whole-cell morphological and organelle statistics between the two cell types. Additionally, we characterize subtle biophysical differences between individual insulin and glucagon vesicles by analyzing vesicle size and molecular density distributions, which were not previously possible using other methods. These sub-vesicular parameters enable us to predict cell types systematically using supervised machine learning. We also visualize distinct vesicle and cell subtypes using Uniform Manifold Approximation and Projection (UMAP) embeddings, which provides us with an innovative approach to explore structural heterogeneity in islet cells. This methodology presents an innovative approach for tracking biologically meaningful heterogeneity in cells that can be applied to any cellular system. Full article
(This article belongs to the Special Issue Advanced Technology for Cellular Imaging)
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Graphical abstract

11 pages, 3511 KiB  
Communication
Survivin-Sodium Iodide Symporter Reporter as a Non-Invasive Diagnostic Marker to Differentiate Uterine Leiomyosarcoma from Leiomyoma
by Natalia Garcia, Mara Ulin, Qiwei Yang, Mohamed Ali, Maarten C. Bosland, Weiqiao Zeng, Liaohai Chen and Ayman Al-Hendy
Cells 2023, 12(24), 2830; https://doi.org/10.3390/cells12242830 - 13 Dec 2023
Cited by 1 | Viewed by 1323
Abstract
Leiomyosarcoma (LMS) has been challenging to diagnose because of limitations in clinical and radiographic predictors, as well as the lack of reliable serum or urinary biomarkers. Most uterine masses consist of benign leiomyoma (LM). However, it is currently a significant challenge in gynecology [...] Read more.
Leiomyosarcoma (LMS) has been challenging to diagnose because of limitations in clinical and radiographic predictors, as well as the lack of reliable serum or urinary biomarkers. Most uterine masses consist of benign leiomyoma (LM). However, it is currently a significant challenge in gynecology practice to differentiate LMS from LM. This inability poses grave consequences for patients, leading to a high number of unnecessary hysterectomies, infertility, and other major morbidities and possible mortalities. This study aimed to evaluate the use of Survivin-Sodium iodide symporter (Ad-Sur-NIS) as a reporter gene biomarker to differentiate malignant LMS from benign LM by using an F18-NaBF4 PET/CT scan. The PET/CT scan images showed a significantly increased radiotracer uptake and a decreased radiotracer decay attributable to the higher abundance of Ad-Sur-NIS in the LMS tumors compared to LM (p < 0.05). An excellent safety profile was observed, with no pathological or metabolic differences detected in Ad-Sur-NIS-treated animal versus the vehicle control. Ad-Sur-NIS as a PET scan reporter is a promising imaging biomarker that can differentiate uterine LMS from LM using F18-NaBF4 as a radiotracer. As a new diagnostic method, the F18 NaBF4 PET/CT scan can provide a much-needed tool in clinical practices to effectively triage women with suspicious uterine masses and avoid unnecessary invasive interventions. Full article
(This article belongs to the Special Issue Advanced Technology for Cellular Imaging)
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