Search

Your search keyword '"Single-cell scale"' showing total 5 results
Start Over Descriptor "Single-cell scale"
5 results on '"Single-cell scale"'

1. [The Advance of the HSCT Evaluation at Single-Cell Scale --Review].

Author

Tang J, Li XT, Li PL, and Zhu H

Abstract

Hematopoietic stem cell transplantation (HSCT) is one of the effective options for the treatment of irradiation-induced injury on hematopoiesis, malignant hematological diseases, and numerous benign severe hematopathy. However, the cellular composition of the graft for HSCT, as well as the significant events of transplanted HSCs in receipients including HSC homing, engraftment, differentiation, remains to be further elucidated. In recent years, with advances in single-cell techniques, the hematopoiesis has been decoding at single cell scale. In addition, single-cell RNA sequencing (scRNA-seq) has been used in the evaluation of hematopoietic dynamics post HSCT, which may be helpful to improve HSCT protocols and clinical outcomes. Hence, the recent advances of evaluating HSCT at single cell scale and the directions worthy paying attention to in the field have been reviewed briefly.

Published
2023
Full Text
View/download PDF

2. Temporal variation of recombinant protein expression in Escherichia coli biofilms analysed at single-cell level.

Author

Gomes, L.C., Carvalho, D., Briandet, R., and Mergulhão, F.J.

Subjects
*RECOMBINANT proteins, *ESCHERICHIA coli physiology, *PROTEIN expression, *BIOFILMS, *GREEN fluorescent protein genetics, *HETEROGENEITY, *PHYSIOLOGY
Abstract

Bioprocesses based on surface-associated microorganisms are emerging in environmental and industrial areas due to the physiological specificities and heterogeneities of biofilm cells. This study describes a simple and accurate method for evaluating the recombinant protein expression at a single-cell scale during Escherichia coli biofilm development. The model recombinant protein used was enhanced the green fluorescent protein (eGFP), as its intrinsic fluorescence allows us to quantify expression at both population and single-cell levels. The specific cell fluorescence intensity sharply increased during the first 4 days of biofilm cultivation. Thereafter, it decreased abruptly reaching a low-level plateau until the end of the experiment. During biofilm development, the population became increasingly heterogeneous with regard to eGFP expression. Three distinct biofilm types were observed over the course of the experiment: one with a homogeneous population (days 3–5), the second with a moderately heterogeneous population (days 6–8) and the third with a strongly heterogeneous population (days 9–11). Observation of E. coli biofilms by confocal laser scanning microscopy revealed marked spatial heterogeneity, with the cells actively producing eGFP restricted to the top layer of the biofilm. The proposed methodology allows a fine analysis of the recombinant protein expression within E. coli biofilms, and it may be used to optimize the processing conditions. [ABSTRACT FROM AUTHOR]

Published
2016
Full Text
View/download PDF

3. Synchrotron FTIR microspectroscopy of Escherichia coli at single-cell scale under silver-induced stress conditions.

Author

Saulou, Claire, Jamme, Frédéric, Girbal, Laurence, Maranges, Claude, Fourquaux, Isabelle, Cocaign-Bousquet, Muriel, Dumas, Paul, and Mercier-Bonin, Muriel

Subjects
*ESCHERICHIA coli, *SILVER, *FATTY acids, *PROTEIN research, *MATHEMATICAL models, *DYNAMICS
Abstract

The present work was focused on elucidating biochemical changes in the model bacterium Escherichia coli exposed to ionic silver mediated stress, at a single-cell scale. In order to achieve this, in situ synchrotron Fourier-transform infrared (sFTIR) microspectroscopy was performed, for the first time, on individual cells by attenuated total reflectance (ATR) combined with the use of zinc-selenide hemisphere for high spatial resolution. In a first part, the potential of the method was evaluated on bacteria subjected to a lethal 100 μM AgNO concentration for 2 h compared to untreated 100 % viable cells. Differences in cell composition were assessed for the C-H stretching and protein spectral regions, indicating that the inhibitory action was targeted against both fatty acids and proteins. Transmission electron microscopy (TEM) confirmed morphological damages of the cell ultrastructure. The relevance of ATR-sFTIR microspectroscopy for highlighting the heterogeneity in Ag-mediated effects within a given bacterial population was also pointed out. In a second part, cells were exposed to sub-lethal Ag concentrations (<10 μM AgNO) tested under 'dynamic' growth mode: early addition vs. pulse in the mid-exponential phase, and compared to simultaneously batch-grown untreated bacteria or cells sampled just before the pulse, respectively. sFTIR microspectroscopy and TEM imaging were performed in close relation with growth kinetics characterization. No significant effect of the Ag pulses was detected, in accordance with macrokinetics data. For early-treated cells, effects on fatty acid composition were shown, although no major alteration of protein secondary structure was noticed. These partial effects were consistent with TEM observations and growth kinetics. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Published
2013
Full Text
View/download PDF

4. Temporal variation of recombinant protein expression in Escherichia coli biofilms analysed at single-cell level

Author

Daniel T. O. Carvalho, Filipe Mergulhão, Romain Briandet, Luciana C. Gomes, LEPABE - Department of Chemical Engineering, Faculty of Engineering, Universidade do Porto, LEPABE – Department of Chemical Engineering, Faculty of Engineering, MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Laboratory for Process Engineering, Environment, Biotechnology and Energy-LEPABE - FEDER funds through COMPETE, Programa Operacional Coinpetitividade e Internacionalizacao (POCI) [POCI-01-0145-FEDER-006939], national funds through FCT-Fundacao para a Ciencia e a Tecnologia [SFRH/BD/80400/2011], FEMS (Federation of European Microbiological Societies), Universidade do Porto [Porto], and Faculdade de Engenharia

Subjects
0301 basic medicine, Microorganism, [SDV]Life Sciences [q-bio], 030106 microbiology, Population, Bioengineering, Biology, Recombinant protein expression, medicine.disease_cause, Applied Microbiology and Biotechnology, Biochemistry, Protein expression, law.invention, Green fluorescent protein, 03 medical and health sciences, law, Fluorescence microscope, medicine, Escherichia coli, education, Fluorescence microscopy, education.field_of_study, Biofilm, Single-cell scale, Molecular biology, Recombinant DNA, Biophysics, Spatial heterogeneity
Abstract

Bioprocesses based on surface-associated microorganisms are emerging in environmental and industrial areas due to the physiological specificities and heterogeneities of biofilm cells. This study describes a simple and accurate method for evaluating the recombinant protein expression at a single-cell scale during Escherichia coli biofilm development. The model recombinant protein used was enhanced the green fluorescent protein (eGFP), as its intrinsic fluorescence allows us to quantify expression at both population and single-cell levels. The specific cell fluorescence intensity sharply increased during the first 4 days of biofilm cultivation. Thereafter, it decreased abruptly reaching a low-level plateau until the end of the experiment. During biofilm development, the population became increasingly heterogeneous with regard to eGFP expression. Three distinct biofilm types were observed over the course of the experiment: one with a homogeneous population (days 3–5), the second with a moderately heterogeneous population (days 6–8) and the third with a strongly heterogeneous population (days 9–11). Observation of E. coli biofilms by confocal laser scanning microscopy revealed marked spatial heterogeneity, with the cells actively producing eGFP restricted to the top layer of the biofilm. The proposed methodology allows a fine analysis of the recombinant protein expression within E. coli biofilms, and it may be used to optimize the processing conditions.

Published
2016

5. Synchrotron FTIR microspectroscopy of Escherichia coli at single-cell scale under silver-induced stress conditions

Author

Claire Saulou, Muriel Mercier-Bonin, Isabelle Fourquaux, Paul Dumas, Claude Maranges, Frédéric Jamme, Muriel Cocaign-Bousquet, Laurence Girbal, Génie et Microbiologie des Procédés Alimentaires (GMPA), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), Unité de recherche sur les Biopolymères, Interactions Assemblages (BIA), Institut National de la Recherche Agronomique (INRA), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT), Centre de Microscopie Électronique Appliquée à la Biologie (CMEAB), Toulouse Réseau Imagerie-Genotoul ( TRI-Genotoul), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), The authors gratefully acknowledge the 'BioPleasure' project of the Research National Agency (ANR-07-BLAN-0196-01) for funding., The research described in this paper was performed at the French national synchrotron facility SOLEIL (Gif-sur- Yvette, France), using the SMIS beamline (proposal no 20090556)., ANR-07-BLAN-0196,BIOPLEASURE,Plasma - surface engineering for biofilm prevention(2007), AgroParisTech-Institut National de la Recherche Agronomique (INRA), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées, Institut National des Sciences Appliquées (INSA), Hôpital de Rangueil, CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Toulouse Réseau Imagerie-Genotoul ( TRI-Genotoul), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Hôpital de Rangueil, and CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]

Subjects
In situ, Analytical chemistry, medicine.disease_cause, Biochemistry, Analytical Chemistry, 03 medical and health sciences, Stress, Physiological, Spectroscopy, Fourier Transform Infrared, medicine, Escherichia coli, Fourier transform infrared spectroscopy, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Protein secondary structure, 030304 developmental biology, 0303 health sciences, biology, Ionic silver, 030306 microbiology, Chemistry, Synchrotron FTIR-ATR microspectroscopy, Single-cell scale, [CHIM.MATE]Chemical Sciences/Material chemistry, biology.organism_classification, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Antibacterial effect, Transmission electron microscopy, Attenuated total reflection, Ultrastructure, Biophysics, Silver Nitrate, Single-Cell Analysis, [SPI.SIGNAL]Engineering Sciences [physics]/Signal and Image processing, Synchrotrons, Bacteria
Abstract

International audience; The present work was focused on elucidating biochemical changes in the model bacterium Escherichia coli exposed to ionic silver mediated stress, at a single-cell scale. In order to achieve this, in situ synchrotron Fourier-transform infrared (sFTIR) microspectroscopy was performed, for the first time, on individual cells by attenuated total reflectance (ATR) combined with the use of zinc-selenide hemisphere for high spatial resolution. In a first part, the potential of the method was evaluated on bacteria subjected to a lethal 100 μM AgNO3 concentration for 2 h compared to untreated 100 % viable cells. Differences in cell composition were assessed for the C–H stretching and protein spectral regions, indicating that the inhibitory action was targeted against both fatty acids and proteins. Transmission electron microscopy (TEM) confirmed morphological damages of the cell ultrastructure. The relevance of ATR-sFTIR microspectroscopy for highlighting the heterogeneity in Ag+-mediated effects within a given bacterial population was also pointed out. In a second part, cells were exposed to sub-lethal Ag+ concentrations (

Published
2013

Catalog

Books, media, physical & digital resources
See catalog results