Live imaging of fluorescent proteins in chordate embryos: From ascidians to mice

2006 ◽  
Vol 69 (3) ◽  
pp. 160-167 ◽  
Author(s):  
Yale J. Passamaneck ◽  
Anna Di Gregorio ◽  
Virginia E. Papaioannou ◽  
Anna-Katerina Hadjantonakis
Keyword(s):  
Fluorescent Proteins ◽  
Live Imaging

scholarly journals Live-imaging fluorescent proteins in mouse embryos: multi-dimensional, multi-spectral perspectives

2009 ◽  
Vol 27 (5) ◽  
pp. 266-276 ◽  
Author(s):  
Sonja Nowotschin ◽  
Guy S. Eakin ◽  
Anna-Katerina Hadjantonakis
Keyword(s):  
Fluorescent Proteins ◽  
Live Imaging ◽  
Mouse Embryos

Illuminating subcellular structures and dynamics in plants: a fluorescent protein toolboxThis review is one of a selection of papers published in the Special Issue on Plant Cell Biology.

2006 ◽  
Vol 84 (4) ◽  
pp. 515-522 ◽  
Author(s):  
Preetinder K. Dhanoa ◽  
Alison M. Sinclair ◽  
Robert T. Mullen ◽  
Jaideep Mathur
Keyword(s):  
Plant Cell ◽  
Cell Biology ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Live Imaging ◽  
Living Cells ◽  
Special Issue ◽  
Exciting Possibility ◽  
Selection Of

The discovery and development of multicoloured fluorescent proteins has led to the exciting possibility of observing a remarkable array of subcellular structures and dynamics in living cells. This minireview highlights a number of the more common fluorescent protein probes in plants and is a testimonial to the fact that the plant cell has not lagged behind during the live-imaging revolution and is ready for even more in-depth exploration.

scholarly journals Development of the Middle Layer in the Anther of Arabidopsis

2021 ◽  
Vol 12 ◽  
Author(s):  
Jing-Shi Xue ◽  
Chi Yao ◽  
Qin-Lin Xu ◽  
Chang-Xu Sui ◽  
Xin-Lei Jia ◽  
...  
Keyword(s):  
Cell Division ◽  
Fluorescent Proteins ◽  
Middle Layer ◽  
Live Imaging ◽  
Anther Development ◽  
Anther Wall ◽  
Fundamental Information ◽  
Stage 4 ◽  
Cell Layers ◽  
Staining Methods

The middle layer is an essential cell layer of the anther wall located between the endothecium and tapetum in Arabidopsis. Based on sectioning, the middle layer was found to be degraded at stage 7, which led to the separation of the tapetum from the anther wall. Here, we established techniques for live imaging of the anther. We created a marker line with fluorescent proteins expressed in all anther layers to study anther development. Several staining methods were used in the intact anthers to study anther cell morphology. We clarified the initiation, development, and degradation of the middle layer in Arabidopsis. This layer is initiated from both the inner and outer secondary parietal cells at stage 4, stopped cell division at stage 6, and finally degraded at stage 11. The neighboring cell layers, the epidermis, and endothecium continued cell division until stage 10, which led to a thin middle layer. The degradation of the tapetum cell wall at stage 7 lead to its isolation from the anther wall. This work presents fundamental information on the development of the middle layer, which facilitates the further investigation of anther development and plant fertility. These live imaging methods could be useful in future studies.

scholarly journals Conditional immobilization for live imaging C. elegans using auxin-dependent protein depletion

2021 ◽  
Author(s):  
Cori K. Cahoon ◽  
Diana E. Libuda
Keyword(s):  
Candidate Genes ◽  
Fluorescent Proteins ◽  
Model Organism ◽  
Live Imaging ◽  
Similar Amount ◽  
Sexually Dimorphic ◽  
Imaging Studies ◽  
C Elegans ◽  
Dependent Protein ◽  
Living Organisms

The visualization of biological processes using fluorescent proteins and dyes in living organisms has enabled numerous scientific discoveries. The nematode Caenorhabditis elegans is a widely used model organism for live imaging studies since the transparent nature of the worm enables imaging of nearly all tissues within a whole, intact animal. While current techniques are optimized to enable the immobilization of hermaphrodite worms for live imaging, many of these approaches fail to successfully restrain the smaller male worms. To enable live imaging of worms of both sexes, we developed a new genetic, conditional immobilization tool that uses the auxin inducible degron (AID) system to immobilize both hermaphrodites and male worms for live imaging. Based on chromosome location, mutant phenotype, and predicted germline consequence, we identified and AID-tagged three candidate genes (unc-18, unc-104, and unc-52). Strains with these AID-tagged genes were placed on auxin and tested for mobility and germline defects. Among the candidate genes, auxin-mediated depletion of UNC-18 caused significant immobilization of both hermaphrodite and male worms that was also partially reversible upon removal from auxin. Notably, we found that male worms require a higher concentration of auxin for a similar amount of immobilization as hermaphrodites, thereby suggesting a potential sex-specific difference in auxin absorption and/or processing. In both males and hermaphrodites, depletion of UNC-18 did not largely alter fertility, germline progression, nor meiotic recombination. Finally, we demonstrate that this new genetic tool can successfully immobilize both sexes enabling live imaging studies of sexually dimorphic features in C. elegans.

scholarly journals Conditional immobilization for live imaging C. elegans using auxin-dependent protein depletion

2021 ◽  
Author(s):  
Cori K Cahoon ◽  
Diana E Libuda
Keyword(s):  
Candidate Genes ◽  
Fluorescent Proteins ◽  
Model Organism ◽  
Live Imaging ◽  
Similar Amount ◽  
Sexually Dimorphic ◽  
Imaging Studies ◽  
C Elegans ◽  
Dependent Protein ◽  
Living Organisms

Abstract The visualization of biological processes using fluorescent proteins and dyes in living organisms has enabled numerous scientific discoveries. The nematode Caenorhabditis elegans is a widely used model organism for live imaging studies since the transparent nature of the worm enables imaging of nearly all tissues within a whole, intact animal. While current techniques are optimized to enable the immobilization of hermaphrodite worms for live imaging, many of these approaches fail to successfully restrain the smaller male worms. To enable live imaging of worms of both sexes, we developed a new genetic, conditional immobilization tool that uses the auxin inducible degron (AID) system to immobilize both adult and larval hermaphrodite and male worms for live imaging. Based on chromosome location, mutant phenotype, and predicted germline consequence, we identified and AID-tagged three candidate genes (unc-18, unc-104, and unc-52). Strains with these AID-tagged genes were placed on auxin and tested for mobility and germline defects. Among the candidate genes, auxin-mediated depletion of UNC-18 caused significant immobilization of both hermaphrodite and male worms that was also partially reversible upon removal from auxin. Notably, we found that male worms require a higher concentration of auxin for a similar amount of immobilization as hermaphrodites, thereby suggesting a potential sex-specific difference in auxin absorption and/or processing. In both males and hermaphrodites, depletion of UNC-18 did not largely alter fertility, germline progression, nor meiotic recombination. Finally, we demonstrate that this new genetic tool can successfully immobilize both sexes enabling live imaging studies of sexually dimorphic features in C. elegans.

scholarly journals Visual Barcodes for Multiplexing Live Microscopy-Based Assays

2020 ◽  
Author(s):  
Tom Kaufman ◽  
Erez Nitzan ◽  
Nir Firestein ◽  
Miriam Ginzberg ◽  
Seshu Iyengar ◽  
...  
Keyword(s):  
Fluorescent Proteins ◽  
Single Cells ◽  
Live Imaging ◽  
Live Cells ◽  
Balance Growth ◽  
Complex Biological System ◽  
Clonal Identity

Abstract While multiplexing samples using DNA barcoding revolutionized the pace of biomedical discovery, multiplexing of live imaging-based applications has been limited by the number of fluorescent proteins that can be deconvoluted using common microscopy equipment. To address this limitation we developed visual barcodes that discriminate the clonal identity of single cells by targeting different fluorescent proteins to specific subcellular locations. We demonstrate that deconvolution of these barcodes is highly accurate and robust to many cellular perturbations. We then used visual barcodes to generate ‘Signalome’ cell-lines by multiplexing live reporters to monitor the simultaneous activity in 12 branches of signaling, in live cells, at single cell resolution, over time. Using the ‘Signalome’ we identified two distinct clusters of signaling pathways that balance growth and proliferation, emphasizing the importance of growth homeostasis as a central organizing principle in cancer signaling. The ability to multiplex samples in live imaging applications, both in vitro and in vivo may allow better high-content characterization of complex biological system

scholarly journals mBeRFP, a versatile fluorescent tool to enhance multichannel live imaging and its applications

2022 ◽  
Author(s):  
Emmanuel Martin ◽  
Magali Suzanne
Keyword(s):  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Stokes Shift ◽  
Live Imaging ◽  
Red Fluorescent Protein ◽  
Red Fluorescent Proteins ◽  
Living Organisms ◽  
Illumination Source

Cell and developmental biology increasingly require live imaging of protein dynamics in cells, tissues or living organisms. Thanks to the discovery and the development of a panel of fluorescent proteins over the last decades, live imaging has become a powerful and commonly used approach. However, multicolor live imaging remains challenging. The generation of long Stokes shift red fluorescent proteins, such as mBeRFP, offers interesting new perspectives to bypass this limitation. Here, we constructed a set of mBeRFP-expressing vectors and provided a detailed characterization of this fluorescent protein for in vivo live imaging and its applications in Drosophila. Briefly, we showed that a single illumination source is sufficient to simultaneously stimulate mBeRFP and GFP. We demonstrated that mBeRFP can be easily combined with classical green and red fluorescent protein without any crosstalk. We also showed that the low photobleaching of mBeRFP is suitable for live imaging, and that this protein can be used for quantitative applications such as FRAP or laser ablation. Finally, we believe that this fluorescent protein, with the set of new possibilities it offers, constitutes an important tool for cell, developmental and mechano biologists in their current research.

scholarly journals Novel Live Imaging Method Applying Molecular Switching Mechanisms with Fluorescent Proteins

2009 ◽  
Vol 58 (6) ◽  
pp. 447-460 ◽  
Author(s):  
Michio Tomura ◽  
Junko Tanaka ◽  
Osami Kanagawa ◽  
Yoshihiro Miwa
Keyword(s):  
Fluorescent Proteins ◽  
Live Imaging ◽  
Imaging Method ◽  
Molecular Switching
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