Advances and Application of Super-resolution Optical Microscopy

A special issue of Photonics (ISSN 2304-6732).

Deadline for manuscript submissions: closed (15 October 2022) | Viewed by 8941

Special Issue Editors


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Guest Editor
Biophysical Immunology Lab, Kennedy Institute of Rheumatology, University of Oxford, Oxford OX3 7FY, UK
Interests: super-resolution microscopy; structured illumination microscopy; fluorescence microscopy; image analysis; high spatial–temporal resolution microscopy for life sciences
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Guest Editor
MRC Human Immunology Unit and Wolfson Imaging Centre, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Headley Way, Oxford OX3 9DS, UK
Interests: fluorescence microscopy; STED microscopy; advanced microscopy techniques in living cells; development of bespoken microscopy
Special Issues, Collections and Topics in MDPI journals

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Guest Editor
Department of Physics E. Fermi, University of Pisa, Largo B. Pontecorvo 3, 56127 Pisa, Italy
Interests: super-resolution microscopy; biophysics; advanced fluorescence microscopy for life sciences; single-molecule localization microscopy; light-sheet fluorescence microscopy

Special Issue Information

Dear Colleagues,

Over the past two decades, super-resolution microscopy techniques made their way from highly specialized laboratories to turn-key commercial systems widespread trough research facilities. A multitude of novel approaches have been developed to improve performances and make advanced optical microscopes flexible for diverse biological applications and more accessible to general users. However, high-resolution techniques still require specialized knowledge from the user side to properly address biological problems of interest.

With this issue, we are aiming to reduce the gap between microscopy developers and day-to-day users. We will focus on tips and tricks from sample preparation to image acquisition, quality control, data processing and analysis to achieve the best performances and accurate results. This Special Issue targets to gather recent advances in super-resolution microscopy together with its biological applications paving the way to complex biological studies at an unseen level of information.

Dr. Kseniya Korobchevskaya
Dr. Silvia Galiani
Dr. Francesca Cella Zanacchi
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. Photonics is an international peer-reviewed open access monthly 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 2400 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

  • super-resolution microscopy
  • fluorescence microscopy
  • image analysis
  • single-molecule localization microscopy
  • STED
  • SIM
  • structured illumination microscopy
  • enhanced resolution
  • advanced microscopy

Published Papers (3 papers)

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Research

15 pages, 13392 KiB  
Article
Flexible Multiplane Structured Illumination Microscope with a Four-Camera Detector
by Karl A. Johnson, Daniel Noble, Rosa Machado, Tristan C. Paul and Guy M. Hagen
Photonics 2022, 9(7), 501; https://doi.org/10.3390/photonics9070501 - 20 Jul 2022
Cited by 5 | Viewed by 2393
Abstract
Fluorescence microscopy provides an unparalleled tool for imaging biological samples. However, producing high-quality volumetric images quickly and without excessive complexity remains a challenge. Here, we demonstrate a four-camera structured illumination microscope (SIM) capable of simultaneously imaging multiple focal planes, allowing for the capture [...] Read more.
Fluorescence microscopy provides an unparalleled tool for imaging biological samples. However, producing high-quality volumetric images quickly and without excessive complexity remains a challenge. Here, we demonstrate a four-camera structured illumination microscope (SIM) capable of simultaneously imaging multiple focal planes, allowing for the capture of 3D fluorescent images without any axial movement of the sample. This setup allows for the acquisition of many different 3D imaging modes, including 3D time lapses, high-axial-resolution 3D images, and large 3D mosaics. We imaged mitochondrial motions in live cells, neuronal structure in Drosophila larvae, and imaged up to 130 µm deep into mouse brain tissue. After SIM processing, the resolution measured using one of the four cameras improved from 357 nm to 253 nm when using a 30×/1.05 NA objective. Full article
(This article belongs to the Special Issue Advances and Application of Super-resolution Optical Microscopy)
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10 pages, 21938 KiB  
Article
Exploration of Deformation of F-Actin during Macropinocytosis by Confocal Microscopy and 3D-Structured Illumination Microscopy
by Linyu Xu, Yanwei Zhang, Song Lang and Yan Gong
Photonics 2022, 9(7), 461; https://doi.org/10.3390/photonics9070461 - 30 Jun 2022
Cited by 1 | Viewed by 1818
Abstract
Since their invention, confocal microscopy and super-resolution microscopy have become important choices in cell biology research. Macropinocytosis is a critical form of endocytosis. Deformation of the cell membrane is thought to be closely related to the movement of F-actin during macropinocytosis. However, it [...] Read more.
Since their invention, confocal microscopy and super-resolution microscopy have become important choices in cell biology research. Macropinocytosis is a critical form of endocytosis. Deformation of the cell membrane is thought to be closely related to the movement of F-actin during macropinocytosis. However, it is still unclear how the morphology of F-actin and the membrane change during this process. In this study, confocal microscopy was utilized for macroscopic time-series imaging of the cell membranes and F-actin in cells induced by phorbol 12-myristate 13-acetate (PMA). Super-resolution structured illumination microscopy (SIM), which can overcome the diffraction limit, was used to demonstrate the morphological characteristics of F-actin filaments. Benefiting from the advantages of SIM in terms of resolution and 3D imaging, we speculated on the regular pattern of the deformation of F-actin during macropinocytosis. The detailed visualization of structures also helped to validate the speculation regarding the role of F-actin filaments in macropinocytosis in previous studies. The results obtained in this study will provide a better understanding of the mechanisms underlying macropinocytosis and endocytosis. Full article
(This article belongs to the Special Issue Advances and Application of Super-resolution Optical Microscopy)
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13 pages, 25551 KiB  
Article
Single-Molecule Clustering for Super-Resolution Optical Fluorescence Microscopy
by Prakash Joshi and Partha Pratim Mondal
Photonics 2022, 9(1), 7; https://doi.org/10.3390/photonics9010007 - 24 Dec 2021
Cited by 3 | Viewed by 3781
Abstract
Molecular assembly in a complex cellular environment is vital for understanding underlying biological mechanisms. Biophysical parameters (such as single-molecule cluster density, cluster-area, pairwise distance, and number of molecules per cluster) related to molecular clusters directly associate with the physiological state (healthy/diseased) of a [...] Read more.
Molecular assembly in a complex cellular environment is vital for understanding underlying biological mechanisms. Biophysical parameters (such as single-molecule cluster density, cluster-area, pairwise distance, and number of molecules per cluster) related to molecular clusters directly associate with the physiological state (healthy/diseased) of a cell. Using super-resolution imaging along with powerful clustering methods (K-means, Gaussian mixture, and point clustering), we estimated these critical biophysical parameters associated with dense and sparse molecular clusters. We investigated Hemaglutinin (HA) molecules in an Influenza type A disease model. Subsequently, clustering parameters were estimated for transfected NIH3T3 cells. Investigations on test sample (randomly generated clusters) and NIH3T3 cells (expressing Dendra2-Hemaglutinin (Dendra2-HA) photoactivable molecules) show a significant disparity among the existing clustering techniques. It is observed that a single method is inadequate for estimating all relevant biophysical parameters accurately. Thus, a multimodel approach is necessary in order to characterize molecular clusters and determine critical parameters. The proposed study involving optical system development, photoactivable sample synthesis, and advanced clustering methods may facilitate a better understanding of single molecular clusters. Potential applications are in the emerging field of cell biology, biophysics, and fluorescence imaging. Full article
(This article belongs to the Special Issue Advances and Application of Super-resolution Optical Microscopy)
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