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Article ; Online: Targeted Laser Ablation in the Embryo of Saccharina latissima.

Boscq, Samuel / Dutertre, Stéphanie / Theodorou, Ioannis / Charrier, Bénédicte

Journal of visualized experiments : JoVE

2022 , Issue 181

Abstract: In Saccharina latissima, the embryo develops as a monolayered cell sheet called the lamina or the blade. Each embryo cell is easy to observe, readily distinguishable from its neighbors, and can be individually targeted. For decades, laser ablation has ... ...

Abstract	In Saccharina latissima, the embryo develops as a monolayered cell sheet called the lamina or the blade. Each embryo cell is easy to observe, readily distinguishable from its neighbors, and can be individually targeted. For decades, laser ablation has been used to study embryo development. Here, a protocol for cell-specific laser ablation was developed for early embryos of the brown alga S. latissima. The presented work includes: (1) the preparation of Saccharina embryos, with a description of the critical parameters, including culture conditions, (2) the laser ablation settings, and (3) the monitoring of the subsequent growth of the irradiated embryo using time-lapse microscopy. In addition, details are provided on the optimal conditions for transporting the embryos from the imaging platform back to the lab, which can profoundly affect subsequent embryo development. Algae belonging to the order Laminariales display embryogenesis patterns similar to Saccharina; this protocol can thus be easily transferred to other species in this taxon.
MeSH term(s)	Embryo, Mammalian ; Embryonic Development ; Laser Therapy ; Phaeophyceae
Language	English
Publishing date	2022-03-11
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/63518
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Targeted laser ablation in the embryo of Saccharina latissima

Boscq, Samuel / Dutertre, Stéphanie / Theodorou, Ioannis / Charrier, Bénédicte

Journal of visualized experiments. 2022 Mar. 11, , no. 181

2022

Abstract	In Saccharina latissima, the embryo develops as a monolayered cell sheet called the lamina or the blade. Each embryo cell is easy to observe, readily distinguishable from its neighbors, and can be individually targeted. For decades, laser ablation has been used to study embryo development. Here, a protocol for cell-specific laser ablation was developed for early embryos of the brown alga S. latissima. The presented work includes: (1) the preparation of Saccharina embryos, with a description of the critical parameters, including culture conditions, (2) the laser ablation settings, and (3) the monitoring of the subsequent growth of the irradiated embryo using time-lapse microscopy. In addition, details are provided on the optimal conditions for transporting the embryos from the imaging platform back to the lab, which can profoundly affect subsequent embryo development. Algae belonging to the order Laminariales display embryogenesis patterns similar to Saccharina; this protocol can thus be easily transferred to other species in this taxon.
Keywords	Saccharina latissima ; embryogenesis ; irradiation ; microscopy
Language	English
Dates of publication	2022-0311
Size	p. e63518.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/63518
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Article: Encapsulation of Luminescent Gold Nanoclusters into Synthetic Vesicles.

Chiechio, Regina M / Ducarre, Solène / Marets, Célia / Dupont, Aurélien / Even-Hernandez, Pascale / Pinson, Xavier / Dutertre, Stéphanie / Artzner, Franck / Musumeci, Paolo / Ravel, Célia / Faro, Maria Jose Lo / Marchi, Valérie

Nanomaterials (Basel, Switzerland)

2022 Volume 12, Issue 21

Abstract: Gold nanoclusters (Au NCs) are attractive luminescent nanoprobes for biomedical applications. In vivo biosensing and bioimaging requires the delivery of the Au NCs into subcellular compartments. In this view, we explore here the possible encapsulation of ...

Abstract	Gold nanoclusters (Au NCs) are attractive luminescent nanoprobes for biomedical applications. In vivo biosensing and bioimaging requires the delivery of the Au NCs into subcellular compartments. In this view, we explore here the possible encapsulation of ultra-small-sized red and blue emitting Au NCs into liposomes of various sizes and chemical compositions. Different methods were investigated to prepare vesicles containing Au NCs in their lumen. The efficiency of the process was correlated to the structural and morphological aspect of the Au NCs' encapsulating vesicles thanks to complementary analyses by SAXS, cryo-TEM, and confocal microscopy techniques. Cell-like-sized vesicles (GUVs) encapsulating red or blue Au NCs were successfully obtained by an innovative method using emulsion phase transfer. Furthermore, exosome-like-sized vesicles (LUVs) containing Au NCs were obtained with an encapsulation yield of 40%, as estimated from ICP-MS.
Language	English
Publishing date	2022-11-02
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662255-5
ISSN	2079-4991
ISSN	2079-4991
DOI	10.3390/nano12213875
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Article ; Online: Encapsulation of Luminescent Gold Nanoclusters into Synthetic Vesicles

Regina M. Chiechio / Solène Ducarre / Célia Marets / Aurélien Dupont / Pascale Even-Hernandez / Xavier Pinson / Stéphanie Dutertre / Franck Artzner / Paolo Musumeci / Célia Ravel / Maria Jose Lo Faro / Valérie Marchi

Nanomaterials, Vol 12, Iss 3875, p

2022 Volume 3875

Abstract	Gold nanoclusters (Au NCs) are attractive luminescent nanoprobes for biomedical applications. In vivo biosensing and bioimaging requires the delivery of the Au NCs into subcellular compartments. In this view, we explore here the possible encapsulation of ultra-small-sized red and blue emitting Au NCs into liposomes of various sizes and chemical compositions. Different methods were investigated to prepare vesicles containing Au NCs in their lumen. The efficiency of the process was correlated to the structural and morphological aspect of the Au NCs’ encapsulating vesicles thanks to complementary analyses by SAXS, cryo-TEM, and confocal microscopy techniques. Cell-like-sized vesicles (GUVs) encapsulating red or blue Au NCs were successfully obtained by an innovative method using emulsion phase transfer. Furthermore, exosome-like-sized vesicles (LUVs) containing Au NCs were obtained with an encapsulation yield of 40%, as estimated from ICP-MS.
Keywords	gold nanoclusters ; vesicles ; encapsulation ; Chemistry ; QD1-999
Subject code	540
Language	English
Publishing date	2022-11-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article: Label-free microscopy of mitotic chromosomes using the polarization orthogonality breaking technique.

Desapogu, Rajesh / Le Marchand, Gilles / Smith, Rebecca / Ray, Paulami / Gillier, Émilie / Dutertre, Stéphanie / Alouini, Mehdi / Tramier, Marc / Huet, Sébastien / Fade, Julien

Biomedical optics express

2021 Volume 12, Issue 8, Page(s) 5290–5304

Abstract: We report how a recently developed polarization imaging technique, implementing micro-wave photonics and referred to as orthogonality-breaking (OB) imaging, can be adapted on a classical confocal fluorescence microscope, and is able to provide ... ...

Abstract	We report how a recently developed polarization imaging technique, implementing micro-wave photonics and referred to as orthogonality-breaking (OB) imaging, can be adapted on a classical confocal fluorescence microscope, and is able to provide informative polarization images from a single scan of the cell sample. For instance, the comparison of the images of various cell lines at different cell-cycle stages obtained by OB polarization microscopy and fluorescence confocal images shows that an endogenous polarimetric contrast arizes with this instrument on compacted chromosomes during cell division.
Language	English
Publishing date	2021-07-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	2572216-5
ISSN	2156-7085
ISSN	2156-7085
DOI	10.1364/BOE.426630
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Article ; Online: Neurite analyzer: An original Fiji plugin for quantification of neuritogenesis in two-dimensional images.

Haas, Alexis J / Prigent, Sylvain / Dutertre, Stéphanie / Le Dréan, Yves / Le Page, Yann

Journal of neuroscience methods

2016 Volume 271, Page(s) 86–91

Abstract: Background: In life sciences, there is a growing need for new informatics tools designed to provide automated solutions in order to analyze big amounts of images obtained from high-throughput imaging systems. Among the most widely used assays in ... ...

Abstract	Background: In life sciences, there is a growing need for new informatics tools designed to provide automated solutions in order to analyze big amounts of images obtained from high-throughput imaging systems. Among the most widely used assays in neurotoxicity, endocrinology and brain diseases, the neurite outgrowth assay is popular. New method: Cell-to-cell quantification of the main morphological features of neurite outgrowth assays remains very challenging. Here, we provide a new pipeline developed on Fiji software for analysis of series of two-dimensional images. It allows the automated analysis of most of these features. Results: We tested the accuracy and usefulness of the software by confirming the effects of estradiol and hypoxia on in vitro neuronal differentiation, previously published by different authors with manual analysis methods. With this new method, we highlighted original interesting data. Comparison with existing method(s): The innovation brought by this plugin lies in the fact that it can process multiple images at the same time, in order to obtain: the number of nuclei, the number of neurites, the length of neurites, the number of neurites junctions, the number of neurites branches, the length of each branch, the position of the branch in the image, the angle of each branch, but also the area of each cell and the number of neurites per cell. Conclusions: This plugin is easy to use, highly sensitive, and allows the experimenter to acquire ready-to-use data coming from a vast amount of images.
MeSH term(s)	Animals ; Cell Hypoxia/physiology ; Estradiol/pharmacology ; Estrogen Receptor alpha/genetics ; Estrogen Receptor alpha/metabolism ; Estrogens/pharmacology ; Image Processing, Computer-Assisted/methods ; Immunohistochemistry/methods ; Microscopy/methods ; Nerve Growth Factor/pharmacology ; Neurites/drug effects ; Neurites/physiology ; Neurogenesis/drug effects ; Neurogenesis/physiology ; Neuronal Outgrowth/drug effects ; Neuronal Outgrowth/physiology ; PC12 Cells ; Pattern Recognition, Automated/methods ; Rats ; Software
Chemical Substances	Estrogen Receptor alpha ; Estrogens ; Estradiol (4TI98Z838E) ; Nerve Growth Factor (9061-61-4)
Language	English
Publishing date	2016-07-20
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	282721-9
ISSN	1872-678X ; 0165-0270
ISSN (online)	1872-678X
ISSN	0165-0270
DOI	10.1016/j.jneumeth.2016.07.011
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Zs.A 1486: Show issues

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Article ; Online: The Ouzo effect: A tool to elaborate high-payload nanocapsules.

Goubault, Clément / Sciortino, Flavien / Mongin, Olivier / Jarry, Ulrich / Bostoën, Mégane / Jakobczyk, Hélène / Burel, Agnès / Dutertre, Stéphanie / Troadec, Marie-Bérengère / Kahn, Myrtil L / Chevance, Soizic / Gauffre, Fabienne

Journal of controlled release : official journal of the Controlled Release Society

2020 Volume 324, Page(s) 430–439

Abstract: We investigate the encapsulation in hybridosomes®, a type of capsules unique regarding their structure and method of elaboration. Hybridosomes® are made of a single shell of inorganic nanoparticles (~5 nm) crosslinked with a polymer and are easily ... ...

Abstract	We investigate the encapsulation in hybridosomes®, a type of capsules unique regarding their structure and method of elaboration. Hybridosomes® are made of a single shell of inorganic nanoparticles (~5 nm) crosslinked with a polymer and are easily obtained via spontaneous emulsification in a ternary mixture THF/water/butylated hydroxytoluene (BHT). Our main finding is that an exceptionally high concentration of a hydrophobic model dye can be loaded in the hybridosomes®, up to 0.35 mol.L
MeSH term(s)	Hydrophobic and Hydrophilic Interactions ; Nanocapsules ; Nanoparticles ; Polymers ; Solvents
Chemical Substances	Nanocapsules ; Polymers ; Solvents
Language	English
Publishing date	2020-05-18
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2020.05.023
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Article: The Ouzo effect: A tool to elaborate high-payload nanocapsules

Journal of controlled release. 2020 Aug. 10, v. 324

2020

Abstract	We investigate the encapsulation in hybridosomes®, a type of capsules unique regarding their structure and method of elaboration. Hybridosomes® are made of a single shell of inorganic nanoparticles (~5 nm) crosslinked with a polymer and are easily obtained via spontaneous emulsification in a ternary mixture THF/water/butylated hydroxytoluene (BHT). Our main finding is that an exceptionally high concentration of a hydrophobic model dye can be loaded in the hybridosomes®, up to 0.35 mol.L⁻¹ or equivalently 170 g.L⁻¹ or 450,000 molecules/capsule. The detailed investigation of the encapsulation mechanism shows that the dye concentrates in the droplets during the emulsification step simultaneously with capsule formation. Then it precipitates inside the capsules during the course of solvent evaporation. In vitro fluorescence measurements show that the nano-precipitated cargo can be transferred from the core of the hybridosomes® to the membrane of liposomes. In vivo studies suggest that the dye diffuses through the body during several days. The released dye tends to accumulate in body-fat, while the inorganic nanoparticles remain trapped into the liver and the spleen macrophages.
Keywords	body fat ; butylated hydroxytoluene ; crosslinking ; droplets ; dyes ; emulsifying ; encapsulation ; evaporation ; fluorescence ; hydrophobicity ; in vivo studies ; liver ; macrophages ; models ; nanocapsules ; nanoparticles ; polymers ; solvents ; spleen
Language	English
Dates of publication	2020-0810
Size	p. 430-439.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2020.05.023
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Poly(ADP-ribose)-dependent chromatin unfolding facilitates the association of DNA-binding proteins with DNA at sites of damage.

Smith, Rebecca / Lebeaupin, Théo / Juhász, Szilvia / Chapuis, Catherine / D'Augustin, Ostiane / Dutertre, Stéphanie / Burkovics, Peter / Biertümpfel, Christian / Timinszky, Gyula / Huet, Sébastien

Nucleic acids research

2019 Volume 47, Issue 21, Page(s) 11250–11267

Abstract: The addition of poly(ADP-ribose) (PAR) chains along the chromatin fiber due to PARP1 activity regulates the recruitment of multiple factors to sites of DNA damage. In this manuscript, we investigated how, besides direct binding to PAR, early chromatin ... ...

Abstract	The addition of poly(ADP-ribose) (PAR) chains along the chromatin fiber due to PARP1 activity regulates the recruitment of multiple factors to sites of DNA damage. In this manuscript, we investigated how, besides direct binding to PAR, early chromatin unfolding events controlled by PAR signaling contribute to recruitment to DNA lesions. We observed that different DNA-binding, but not histone-binding, domains accumulate at damaged chromatin in a PAR-dependent manner, and that this recruitment correlates with their affinity for DNA. Our findings indicate that this recruitment is promoted by early PAR-dependent chromatin remodeling rather than direct interaction with PAR. Moreover, recruitment is not the consequence of reduced molecular crowding at unfolded damaged chromatin but instead originates from facilitated binding to more exposed DNA. These findings are further substantiated by the observation that PAR-dependent chromatin remodeling at DNA lesions underlies increased DNAse hypersensitivity. Finally, the relevance of this new mode of PAR-dependent recruitment to DNA lesions is demonstrated by the observation that reducing the affinity for DNA of both CHD4 and HP1α, two proteins shown to be involved in the DNA-damage response, strongly impairs their recruitment to DNA lesions.
MeSH term(s)	Binding Sites/genetics ; Cells, Cultured ; Chromatin/chemistry ; Chromatin/metabolism ; Chromatin Assembly and Disassembly/physiology ; DNA/metabolism ; DNA Damage/genetics ; DNA-Binding Proteins/metabolism ; Humans ; Nucleic Acid Conformation ; Poly Adenosine Diphosphate Ribose/metabolism ; Poly Adenosine Diphosphate Ribose/physiology ; Protein Binding
Chemical Substances	Chromatin ; DNA-Binding Proteins ; Poly Adenosine Diphosphate Ribose (26656-46-2) ; DNA (9007-49-2)
Language	English
Publishing date	2019-09-30
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	186809-3
ISSN	1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
ISSN (online)	1362-4962 ; 1362-4954
ISSN	0301-5610 ; 0305-1048
DOI	10.1093/nar/gkz820
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Article: Aurora-A overexpression leads to override of the microtubule-kinetochore attachment checkpoint.

Dutertre, Stéphanie / Prigent, Claude

Molecular interventions

2004 Volume 3, Issue 3, Page(s) 127–130

Abstract: The amplification of AURORA-A is frequently observed in specific epithelial cell malignancies. The overexpression of aurora-A, a Ser-Thr kinase known to localize to centrosomes during mitosis, appears to imbue cancerous cells with resistance to spindle- ... ...

Abstract	The amplification of AURORA-A is frequently observed in specific epithelial cell malignancies. The overexpression of aurora-A, a Ser-Thr kinase known to localize to centrosomes during mitosis, appears to imbue cancerous cells with resistance to spindle-checkpoint-targeting drugs such as paclitaxel (Taxol). Indeed, a recent publication by Anand et al. indicates that overexpression of AURORA-A may interfere with spindle-microtubule attachment and disrupt the regulation of the spindle checkpoint by allowing cells with abnormal chromosomal separation to enter anaphase. Thus, the design of new drugs that specifically target aurora-A rather than other checkpoint proteins might alleviate the resistance to Taxol-like clinical therapeutics observed in some tumors.
MeSH term(s)	Anaphase ; Animals ; Apoptosis ; Aurora Kinase A ; Aurora Kinases ; Caenorhabditis elegans ; Calcium-Binding Proteins/metabolism ; Cell Cycle Proteins ; Cell Line ; Centrosome/ultrastructure ; Drosophila melanogaster ; HeLa Cells ; Humans ; Kinetochores/ultrastructure ; Mad2 Proteins ; Metaphase ; Mice ; Microtubules/ultrastructure ; Models, Biological ; Protein Kinases/biosynthesis ; Protein Kinases/physiology ; Protein-Serine-Threonine Kinases ; Repressor Proteins ; Saccharomyces cerevisiae ; Xenopus Proteins
Chemical Substances	Calcium-Binding Proteins ; Cell Cycle Proteins ; MAD2L1 protein, human ; Mad2 Proteins ; Repressor Proteins ; Xenopus Proteins ; Protein Kinases (EC 2.7.-) ; AURKA protein, Xenopus (EC 2.7.11.1) ; Aurka protein, mouse (EC 2.7.11.1) ; Aurora Kinase A (EC 2.7.11.1) ; Aurora Kinases (EC 2.7.11.1) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1)
Language	English
Publishing date	2004-02-27
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2108819-6
ISSN	1543-2548 ; 1534-0384
ISSN (online)	1543-2548
ISSN	1534-0384
DOI	10.1124/mi.3.3.127
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