Titre :

Spectroscopie Raman spontanée et stimulée pour l'analyse des adipocytes

Auteur(s) :

Rougé-Labriet, Hélène [Auteur]
Université de Lille
Synchrotron Radiation for Biomedicine = Rayonnement SynchroTROn pour la Recherche BiomédicalE [STROBE]
Marrow Adiposity & Bone Lab - Adiposité Médullaire et Os - ULR 4490 [MABLab]
Delattre, Jérôme [Auteur]
Université de Lille
Marrow Adiposity & Bone Lab - Adiposité Médullaire et Os - ULR 4490 [MABLab]
Falgayrac, Guillaume [Auteur]
Université de Lille
Marrow Adiposity & Bone Lab - Adiposité Médullaire et Os - ULR 4490 [MABLab]
Lacaze, Magalie [Auteur]
Fédération de Recherche Agrobiosciences, Interactions et Biodiversité [FR AIB]
Tratwal, Josefine [Auteur]
Université de Lausanne = University of Lausanne [UNIL]
Bataclan, Charles [Auteur]
Ecole Polytechnique Fédérale de Lausanne [EPFL]
Bertheaume, Nicolas [Auteur]
Université de Lille
Marrow Adiposity & Bone Lab - Adiposité Médullaire et Os - ULR 4490 [MABLab]
Duponchel, Ludovic [Auteur]
Université de Lille
Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement - UMR 8516 [LASIRE]
Naveiras, Olaia [Auteur]
Ecole Polytechnique Fédérale de Lausanne [EPFL]
Olejnik, Cecile [Auteur]
Marrow Adiposity & Bone Lab - Adiposité Médullaire et Os - ULR 4490 [MABLab]
Faculté de Chirurgie Dentaire
Université de Lille
Pouzet, Cécile [Auteur]
Fédération de Recherche Agrobiosciences, Interactions et Biodiversité [FR AIB]

Titre de la manifestation scientifique :

8th International meeting on Bone Marrow Adiposity

Organisateur(s) de la manifestation scientifique :

BMAS

Ville :

Montreal (Canada)

Pays :

Canada

Date de début de la manifestation scientifique :

2024-09-24

Mot(s)-clé(s) en anglais :

Stimulated Raman imaging
Spontaneous Raman Spectroscopy
Adipocyte
Adipocyte Size
Raman imaging
OP9 cell

Discipline(s) HAL :

Sciences du Vivant [q-bio]
Sciences du Vivant [q-bio]/Biochimie, Biologie Moléculaire/Biologie moléculaire

Résumé en anglais : [en]

Performing in-situ analyses of biomolecules in living cells is crucial in biomedical research. There is growing interest in developing techniques to characterize adipocyte maturation rapidly and non-invasively. Multiple ...
Lire la suite >Performing in-situ analyses of biomolecules in living cells is crucial in biomedical research. There is growing interest in developing techniques to characterize adipocyte maturation rapidly and non-invasively. Multiple methods are available to assess the composition of adipocytes. However, these methods are destructive or invasive.Spontaneous Raman spectroscopy (SpRS) is a label-free method to assess adipocytes in-vitro. This technique provides molecular composition information at a micrometer scale without damaging the sample. The Raman spectrum assesses the molecular composition within the probed micro-volume (~2 µm). SpRS suffers from a low signal-to-noise ratio (SNR), leading to long acquisition times of Raman images. Recent development like Stimulated Raman Spectroscopy (SRS) has improved SNR, enabling fast 3D Raman images (100×100×40 µm). This study compares SpRS and SRS for adipocyte analysis. For this work, lipid composition of a murine bone marrow derived stromal cell line was assessed in two differentiation conditions, either through classical induction of adipogenesis (medium supplemented with dexamethasone, 3-isobutyl-1-methylxanthine, and insulin) or through spontaneous adipogenesis (non-supplemented, serum-containing medium) after 17 days in culture. SpRS were done with an HR800 (HORIBA). SRS were done on a Stellaris 8 CARS/CRS (Leica Microsystems). Both SpRS and SRS allows the analysis of individual lipids droplets down to 2µm (LD) within adipocytes. Raman spectra from SpRS are obtained within the fingerprint region (containing characteristics Raman bands) with a high spectral resolution (0.5 nm). The lipid composition can be imaged (Figure A). LD diameters are evaluated by analysis of the optical image. The tradeoff with SpRS is a long acquisition time for 2D Raman images up to 2 adipocytes (~ 3h/image). SRS analysis allows fast acquisition of 3D Raman images of up to 3 adipocytes within 30 min (Figure B). From 3D images, diameter, volume, number of neighbors and unsaturation ratio are evaluated by image analysis. The tradeoff with SRS is the acquisition of one individual band per image with a low spectral resolution (4 nm). SpRS and SRS analyses found similar results. Classical adipogenic induction led to the preferential accumulation of unsaturated lipid species, as opposed to lower unsaturation ratio on spontaneously differentiated adipocytes coupled to differential lipid droplet size. In conclusion, SpRS and SRS are powerful tools for adipocyte analysis in vitro. SpRS is suitable for detailed spectral analysis while SRS is ideal for faster 3D imaging of multiple cells.Lire moins >

Langue :

Anglais

Comité de lecture :

Oui

Audience :

Internationale

Vulgarisation :

Non

Collections :

• Marrow Adiposity & Bone Lab (MABLab) - ULR 4490

Source :

Harvested from HAL