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Cells
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Imaging in Cell Biology and Development
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Imaging in Cell Biology and Development

A special issue of Cells (ISSN 2073-4409).

Deadline for manuscript submissions: closed (15 October 2012) | Viewed by 68221

Special Issue Editors

Prof. Doug Brooks

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Guest Editor
Mechanisms in Cell Biology and Disease Research Group Leader, School of Pharmacy and Medical Sciences, Sansom Institute for Health Research, City East Campus, University of South Australia, Adelaide, SA 5001, Australia
Interests: cancer biology; cell biology; immunochemistry; live cell imaging
Special Issues, Collections and Topics in MDPI journals
Dr. Tetyana Shandala

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Guest Editor
Mechanisms in Cell Biology and Disease Research Group, School of Pharmacy and Medical Sciences, Sansom Institute for Health Research, City East Campus, University of South Australia, Adelaide, SA 5001, Australia

Special Issue Information

Dear Colleagues,

The visualization of cell biological processes has become an important approach to studying protein structure and function. Roger Tsien's discovery of green fluorescent protein and related fluorophores, their use as tools to study cellular processes using fusion proteins and the associated technological revolution in fluorescence microscopy, have seen the adoption of imaging techniques as a primary tool in every cell biologist's bag of tricks. Live cell imaging techniques are now providing another line of investigation for cell biological processes and molecular studies. The recent advances in super resolution microscopy have also added to the structural detail that can be visualised. The high sensitivity and specificity now attainable with spectral and two photon systems and the capacity to both "gate" and utilise auto and endogenous fluorescence have also added significantly to imaging capacity. At a recent meeting of Australasian cell biologists in Adelaide, a range of imaging techniques were discussed and through hands on practical forums new technologies and industry providers engaged. An international guest from the NIH, Dr Roberto Weigert, provided expert instruction on state of the art live cell and intravital imaging and demonstrated applications for respectively the analysis of in vitro and in vivo systems. In this special guest issue on "Imaging in Cell biology and Development" in the Journal Cells, a series of review articles, technical briefs and research articles are being assembled to reflect some of the available techniques and approaches available to the modern cell biologist and how these can be utilised to resolve cellular mechanism.

Prof. Doug Brooks
Dr. Tetyana Shandala
Guest Editors

Published Papers (7 papers)

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Research

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620 KiB  
Article
A Drosophila Model to Image Phagosome Maturation
by Tetyana Shandala, Chiaoxin Lim, Alexandra Sorvina and Douglas A. Brooks
Cells 2013, 2(2), 188-201; https://doi.org/10.3390/cells2020188 - 26 Mar 2013
Cited by 8 | Viewed by 7978
Abstract
Phagocytosis involves the internalization of extracellular material by invagination of the plasma membrane to form intracellular vesicles called phagosomes, which have functions that include pathogen degradation. The degradative properties of phagosomes are thought to be conferred by sequential fusion with endosomes and lysosomes; [...] Read more.
Phagocytosis involves the internalization of extracellular material by invagination of the plasma membrane to form intracellular vesicles called phagosomes, which have functions that include pathogen degradation. The degradative properties of phagosomes are thought to be conferred by sequential fusion with endosomes and lysosomes; however, this maturation process has not been studied in vivo. We employed Drosophila hemocytes, which are similar to mammalian professional macrophages, to establish a model of phagosome maturation. Adult Drosophila females, carrying transgenic Rab7-GFP endosome and Lamp1-GFP lysosome markers, were injected with E. coli DH5α and the hemocytes were collected at 15, 30, 45 and 60 minutes after infection. In wild-type females, E. coli were detected within enlarged Rab7-GFP positive phagosomes at 15 to 45 minutes after infection; and were also observed in enlarged Lamp1-GFP positive phagolysosomes at 45 minutes. Two-photon imaging of hemocytes in vivo confirmed this vesicle morphology, including enlargement of Rab7-GFP and Lamp1-GFP structures that often appeared to protrude from hemocytes. The interaction of endosomes and lysosomes with E. coli phagosomes observed in Drosophila hemocytes was consistent with that previously described for phagosome maturation in human ex vivo macrophages. We also tested our model as a tool for genetic analysis using 14-3-3e mutants, and demonstrated altered phagosome maturation with delayed E. coli internalization, trafficking and/or degradation. These findings demonstrate that Drosophila hemocytes provide an appropriate, genetically amenable, model for analyzing phagosome maturation ex vivo and in vivo. Full article
(This article belongs to the Special Issue Imaging in Cell Biology and Development)
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458 KiB  
Article
Intravital Microscopy Reveals Differences in the Kinetics of Endocytic Pathways between Cell Cultures and Live Animals
by Andrius Masedunskas, Natalie Porat-Shliom, Kamil Rechache, Myo-Pale' Aye and Roberto Weigert
Cells 2012, 1(4), 1121-1132; https://doi.org/10.3390/cells1041121 - 16 Nov 2012
Cited by 20 | Viewed by 7195
Abstract
Intravital microscopy has enabled imaging of the dynamics of subcellular structures in live animals, thus opening the door to investigating membrane trafficking under physiological conditions. Here, we sought to determine whether the architecture and the environment of a fully developed tissue influences the [...] Read more.
Intravital microscopy has enabled imaging of the dynamics of subcellular structures in live animals, thus opening the door to investigating membrane trafficking under physiological conditions. Here, we sought to determine whether the architecture and the environment of a fully developed tissue influences the dynamics of endocytic processes. To this aim, we imaged endocytosis in the stromal cells of rat salivary glands both in situ and after they were isolated and cultured on a solid surface. We found that the internalization of transferrin and dextran, two molecules that traffic via distinct mechanisms, is substantially altered in cultured cells, supporting the idea that the three dimensional organization of the tissue and the cues generated by the surrounding environment strongly affect membrane trafficking events. Full article
(This article belongs to the Special Issue Imaging in Cell Biology and Development)
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954 KiB  
Article
A Novel Type III Endosome Transmembrane Protein, TEMP
by Rajith N. Aturaliya, Markus C. Kerr and Rohan D. Teasdale
Cells 2012, 1(4), 1029-1044; https://doi.org/10.3390/cells1041029 - 05 Nov 2012
Cited by 1 | Viewed by 7114
Abstract
As part of a high-throughput subcellular localisation project, the protein encoded by the RIKEN mouse cDNA 2610528J11 was expressed and identified to be associated with both endosomes and the plasma membrane. Based on this, we have assigned the name TEMP for Type III [...] Read more.
As part of a high-throughput subcellular localisation project, the protein encoded by the RIKEN mouse cDNA 2610528J11 was expressed and identified to be associated with both endosomes and the plasma membrane. Based on this, we have assigned the name TEMP for Type III Endosome Membrane Protein. TEMP encodes a short protein of 111 amino acids with a single, alpha-helical transmembrane domain. Experimental analysis of its membrane topology demonstrated it is a Type III membrane protein with the amino-terminus in the lumenal, or extracellular region, and the carboxy-terminus in the cytoplasm. In addition to the plasma membrane TEMP was localized to Rab5 positive early endosomes, Rab5/Rab11 positive recycling endosomes but not Rab7 positive late endosomes. Video microscopy in living cells confirmed TEMP's plasma membrane localization and identified the intracellular endosome compartments to be tubulovesicular. Overexpression of TEMP resulted in the early/recycling endosomes clustering at the cell periphery that was dependent on the presence of intact microtubules. The cellular function of TEMP cannot be inferred based on bioinformatics comparison, but its cellular distribution between early/recycling endosomes and the plasma membrane suggests a role in membrane transport. Full article
(This article belongs to the Special Issue Imaging in Cell Biology and Development)
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Review

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1266 KiB  
Review
Actin in Action: Imaging Approaches to Study Cytoskeleton Structure and Function
by Katey K. McKayed and Jeremy C. Simpson
Cells 2013, 2(4), 715-731; https://doi.org/10.3390/cells2040715 - 14 Nov 2013
Cited by 34 | Viewed by 10584
Abstract
The cytoskeleton plays several fundamental roles in the cell, including organizing the spatial arrangement of subcellular organelles, regulating cell dynamics and motility, providing a platform for interaction with neighboring cells, and ultimately defining overall cell shape. Fluorescence imaging has proved to be vital [...] Read more.
The cytoskeleton plays several fundamental roles in the cell, including organizing the spatial arrangement of subcellular organelles, regulating cell dynamics and motility, providing a platform for interaction with neighboring cells, and ultimately defining overall cell shape. Fluorescence imaging has proved to be vital in furthering our understanding of the cytoskeleton, and is now a mainstay technique used widely by cell biologists. In this review we provide an introduction to various imaging modalities used to study focal adhesions and the actin cytoskeleton, and using specific examples we highlight a number of recent studies in animal cells that have advanced our knowledge of cytoskeletal behavior. Full article
(This article belongs to the Special Issue Imaging in Cell Biology and Development)
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2012 KiB  
Review
Fluorescein Derivatives in Intravital Fluorescence Imaging
by Thomas A. Robertson, Florestan Bunel and Michael S. Roberts
Cells 2013, 2(3), 591-606; https://doi.org/10.3390/cells2030591 - 02 Aug 2013
Cited by 47 | Viewed by 15672
Abstract
Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM) enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and [...] Read more.
Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM) enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT)] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples. Full article
(This article belongs to the Special Issue Imaging in Cell Biology and Development)
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699 KiB  
Review
Imaging and Quantitation Techniques for Tracking Cargo along Endosome-to-Golgi Transport Pathways
by Pei Zhi Cheryl Chia and Paul A. Gleeson
Cells 2013, 2(1), 105-123; https://doi.org/10.3390/cells2010105 - 22 Feb 2013
Cited by 10 | Viewed by 10207
Abstract
Recent improvements in the resolution of light microscopy, coupled with the development of a range of fluorescent-based probes, have provided new approaches to dissecting membrane domains and the regulation of membrane trafficking. Here, we review these advances, as well as highlight developments in [...] Read more.
Recent improvements in the resolution of light microscopy, coupled with the development of a range of fluorescent-based probes, have provided new approaches to dissecting membrane domains and the regulation of membrane trafficking. Here, we review these advances, as well as highlight developments in quantitative image analysis and novel unbiased analytical approaches to quantitate protein localization. The application of these approaches to endosomal sorting and endosome-to-Golgi transport is discussed. Full article
(This article belongs to the Special Issue Imaging in Cell Biology and Development)
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599 KiB  
Review
Genetic Systems to Investigate Regulation of Oncogenes and Tumour Suppressor Genes in Drosophila
by Jue Er Amanda Lee, Nicola J. Cranna, Arjun S. Chahal and Leonie M. Quinn
Cells 2012, 1(4), 1182-1196; https://doi.org/10.3390/cells1041182 - 05 Dec 2012
Cited by 1 | Viewed by 8801
Abstract
Animal growth requires coordination of cell growth and cell cycle progression with developmental signaling. Loss of cell cycle control is extremely detrimental, with reduced cycles leading to impaired organ growth and excessive proliferation, potentially resulting in tissue overgrowth and driving tumour initiation. Due [...] Read more.
Animal growth requires coordination of cell growth and cell cycle progression with developmental signaling. Loss of cell cycle control is extremely detrimental, with reduced cycles leading to impaired organ growth and excessive proliferation, potentially resulting in tissue overgrowth and driving tumour initiation. Due to the high level of conservation between the cell cycle machinery of Drosophila and humans, the appeal of the fly model continues to be the means with which we can use sophisticated genetics to provide novel insights into mammalian growth and cell cycle control. Over the last decade, there have been major additions to the genetic toolbox to study development in Drosophila. Here we discuss some of the approaches available to investigate the potent growth and cell cycle properties of the Drosophila counterparts of prominent cancer genes, with a focus on the c-Myc oncoprotein and the tumour suppressor protein FIR (Hfp in flies), which behaves as a transcriptional repressor of c-Myc. Full article
(This article belongs to the Special Issue Imaging in Cell Biology and Development)
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