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

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.

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Archaeal imaging: leading the hunt for new discoveries

Molecular Biology of the Cell ◽

10.1091/mbc.e17-10-0603 ◽

2018 ◽

Vol 29 (14) ◽

pp. 1675-1681 ◽

Cited By ~ 14

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.

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Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2020.588922 ◽

2020 ◽

Vol 8 ◽

Author(s):

Fernanda Ramos-Gomes ◽

Nathalia Ferreira ◽

Alexander Kraupner ◽

Frauke Alves ◽

M. Andrea Markus

Keyword(s):

Live Cell Imaging ◽

Cell Imaging ◽

Ex Vivo ◽

Cell Interactions ◽

Live Cell ◽

Mouse Lung

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Shedding light on biology of bacterial cells

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2015.0499 ◽

2016 ◽

Vol 371 (1707) ◽

pp. 20150499 ◽

Cited By ~ 16

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’.

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Live cell imaging of the hyperthermophilic archaeon Sulfolobus acidocaldarius identifies complementary roles for two ESCRTIII homologues in ensuring a robust and symmetric cell division

10.1101/2020.02.18.953042 ◽

2020 ◽

Cited By ~ 2

Author(s):

Andre Arashiro Pulschen ◽

Delyan R. Mutavchiev ◽

Kim Nadine Sebastian ◽

Jacques Roubinet ◽

Marc Roubinet ◽

...

Keyword(s):

Cell Division ◽

Cell Biology ◽

Live Cell Imaging ◽

Cell Imaging ◽

Halophilic Archaea ◽

Live Cell ◽

Tight Coupling ◽

Dna Compaction ◽

Cellular Processes ◽

Daughter Cells

Live-cell imaging has revolutionized our understanding of dynamic cellular processes in bacteria and eukaryotes. While similar techniques have recently been applied to the study of halophilic archaea, our ability to explore the cell biology of thermophilic archaea is limited, due to the technical challenges of imaging at high temperatures. Here, we report the construction of the Sulfoscope, a heated chamber that enables live-cell imaging on an inverted fluorescent microscope. Using this system combined with thermostable fluorescent probes, we were able to image Sulfolobus cells as they divide, revealing a tight coupling between changes in DNA compaction, segregation and cytokinesis. By imaging deletion mutants, we observe important differences in the function of the two ESCRTIII proteins recently implicated in cytokinesis. The loss of CdvB1 compromises cell division, causing occasional division failures and fusion of the two daughter cells, whereas the deletion of cdvB2 leads to a profound loss of division symmetry, generating daughter cells that vary widely in size and eventually generating ghost cells. These data indicate that DNA separation and cytokinesis are coordinated in Sulfolobus, as is the case in eukaryotes, and that two contractile ESCRTIII polymers perform distinct roles to ensure that Sulfolobus cells undergo a robust and symmetrical division. Taken together, the Sulfoscope has shown to provide a controlled high temperature environment, in which cell biology of Sulfolobus can be studied in unprecedent details.

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