scholarly journals Shedding light on biology of bacterial cells

2016 ◽  
Vol 371 (1707) ◽  
pp. 20150499 ◽  
Author(s):  
Johannes P. Schneider ◽  
Marek Basler
Keyword(s):  
Light Microscopy ◽  
Live Cell Imaging ◽  
Spatial Organization ◽  
Cell Imaging ◽  
Live Cell ◽  
Bacterial Cells ◽  
Resolution Limit ◽  
Detection Techniques ◽  
Living Organisms ◽  
Cellular Components

To understand basic principles of living organisms one has to know many different properties of all cellular components, their mutual interactions but also their amounts and spatial organization. Live-cell imaging is one possible approach to obtain such data. To get multiple snapshots of a cellular process, the imaging approach has to be gentle enough to not disrupt basic functions of the cell but also have high temporal and spatial resolution to detect and describe the changes. Light microscopy has become a method of choice and since its early development over 300 years ago revolutionized our understanding of living organisms. As most cellular components are indistinguishable from the rest of the cellular contents, the second revolution came from a discovery of specific labelling techniques, such as fusions to fluorescent proteins that allowed specific tracking of a component of interest. Currently, several different tags can be tracked independently and this allows us to simultaneously monitor the dynamics of several cellular components and from the correlation of their dynamics to infer their respective functions. It is, therefore, not surprising that live-cell fluorescence microscopy significantly advanced our understanding of basic cellular processes. Current cameras are fast enough to detect changes with millisecond time resolution and are sensitive enough to detect even a few photons per pixel. Together with constant improvement of properties of fluorescent tags, it is now possible to track single molecules in living cells over an extended period of time with a great temporal resolution. The parallel development of new illumination and detection techniques allowed breaking the diffraction barrier and thus further pushed the resolution limit of light microscopy. In this review, we would like to cover recent advances in live-cell imaging technology relevant to bacterial cells and provide a few examples of research that has been possible due to imaging. This article is part of the themed issue ‘The new bacteriology’.

scholarly journals Caught in the Act: Live-Cell Imaging of Plant Meiosis

2021 ◽  
Vol 12 ◽  
Author(s):  
Maria Ada Prusicki ◽  
Martina Balboni ◽  
Kostika Sofroni ◽  
Yuki Hamamura ◽  
Arp Schnittger
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Ex Vivo ◽  
Live Cell ◽  
Cellular Processes ◽  
Short Period ◽  
Molecular Forces ◽  
Cellular Components ◽  
Plant Meiosis ◽  
Imaging Conditions

Live-cell imaging is a powerful method to obtain insights into cellular processes, particularly with respect to their dynamics. This is especially true for meiosis, where chromosomes and other cellular components such as the cytoskeleton follow an elaborate choreography over a relatively short period of time. Making these dynamics visible expands understanding of the regulation of meiosis and its underlying molecular forces. However, the analysis of meiosis by live-cell imaging is challenging; specifically in plants, a temporally resolved understanding of chromosome segregation and recombination events is lacking. Recent advances in live-cell imaging now allow the analysis of meiotic events in plants in real time. These new microscopy methods rely on the generation of reporter lines for meiotic regulators and on the establishment of ex vivo culture and imaging conditions, which stabilize the specimen and keep it alive for several hours or even days. In this review, we combine an overview of the technical aspects of live-cell imaging in plants with a summary of outstanding questions that can now be addressed to promote live-cell imaging in Arabidopsis and other plant species and stimulate ideas on the topics that can be addressed in the context of plant meiotic recombination.

scholarly journals Archaeal imaging: leading the hunt for new discoveries

2018 ◽  
Vol 29 (14) ◽  
pp. 1675-1681 ◽  
Author(s):  
Alexandre W. Bisson-Filho ◽  
Jenny Zheng ◽  
Ethan Garner
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Structural Information ◽  
Eukaryotic Cell ◽  
Live Cell ◽  
Biological Processes ◽  
Cellular Processes ◽  
Archaeal Species ◽  
Eukaryotic Organisms ◽  
Cellular Components

Since the identification of the archaeal domain in the mid-1970s, we have collected a great deal of metagenomic, biochemical, and structural information from archaeal species. However, there is still little known about how archaeal cells organize their internal cellular components in space and time. In contrast, live-cell imaging has allowed bacterial and eukaryotic cell biologists to learn a lot about biological processes by observing the motions of cells, the dynamics of their internal organelles, and even the motions of single molecules. The explosion of knowledge gained via live-cell imaging in prokaryotes and eukaryotes has motivated an ever-improving set of imaging technologies that could allow analogous explorations into archaeal biology. Furthermore, previous studies of essential biological processes in prokaryotic and eukaryotic organisms give methodological roadmaps for the investigation of similar processes in archaea. In this perspective, we highlight a few fundamental cellular processes in archaea, reviewing our current state of understanding about each, and compare how imaging approaches helped to advance the study of similar processes in bacteria and eukaryotes.

Single-molecule live-cell imaging of bacterial DNA repair and damage tolerance

2017 ◽  
Vol 46 (1) ◽  
pp. 23-35 ◽  
Author(s):  
Harshad Ghodke ◽  
Han Ho ◽  
Antoine M. van Oijen
Keyword(s):  
Dna Repair ◽  
Single Molecule ◽  
Live Cell Imaging ◽  
Spatial Organization ◽  
Cell Imaging ◽  
Live Cell ◽  
Live Cells ◽  
Temporal Regulation ◽  
Repair Processes ◽  
Wide Range

Genomic DNA is constantly under threat from intracellular and environmental factors that damage its chemical structure. Uncorrected DNA damage may impede cellular propagation or even result in cell death, making it critical to restore genomic integrity. Decades of research have revealed a wide range of mechanisms through which repair factors recognize damage and co-ordinate repair processes. In recent years, single-molecule live-cell imaging methods have further enriched our understanding of how repair factors operate in the crowded intracellular environment. The ability to follow individual biochemical events, as they occur in live cells, makes single-molecule techniques tremendously powerful to uncover the spatial organization and temporal regulation of repair factors during DNA–repair reactions. In this review, we will cover practical aspects of single-molecule live-cell imaging and highlight recent advances accomplished by the application of these experimental approaches to the study of DNA–repair processes in prokaryotes.

Live cell imaging of bacterial cells: Pyrenoylpyrrole-based fluorescence labeling

2017 ◽  
Vol 90 (4) ◽  
pp. 554-560 ◽  
Author(s):  
Mathiyazhagan Arun Divakar ◽  
Sivakumar Shanmugam
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Fluorescence Labeling ◽  
Bacterial Cells

scholarly journals A live-cell imaging system for visualizing the transport of Marburg virus nucleocapsid-like structures

2019 ◽  
Author(s):  
Yuki Takamatsu ◽  
Takeshi Noda ◽  
Stephan Becker
Keyword(s):  
Live Cell Imaging ◽  
Ebola Virus ◽  
Cell Imaging ◽  
Temporal Dynamics ◽  
Imaging System ◽  
Marburg Virus ◽  
Live Cell ◽  
Infected Cells ◽  
Spatio Temporal ◽  
Living Organisms

AbstractLive-cell imaging is a powerful tool for visualization of the spatio-temporal dynamics of living organisms. Although this technique is utilized to visualize nucleocapsid transport in Marburg virus (MARV)- or Ebola virus-infected cells, the experiments require biosafety level-4 (BSL-4) laboratories, which are restricted to trained and authorized individuals. To overcome this limitation, we developed a live-cell imaging system to visualize MARV nucleocapsid-like structures using fluorescence-conjugated viral proteins, which can be conducted outside BSL-4 laboratories. Our experiments revealed that nucleocapsid-like structures have similar transport characteristics to nucleocapsids observed in MARV-infected cells. This system provides a safe platform to evaluate antiviral drugs that inhibit MARV nucleocapsid transport.

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