Name: Mappatura meccanica degli sferoidi tramite spettroscopia di Brillouin (Video) | JoVE Journal
Uploaded: 2025-12-12T13:10:00.000+00:00
Duration: 8 min 27 s
Description: Impara la caratterizzazione meccanica di sferoidi tumorali in matrice 3D tramite micro-spettroscopia Brillouin. Questo metodo non invasivo consente di studiare la meccanobiologia e la progressione patologica.

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</ol>

<table><tbody><tr><td>PEG-Maleimide a 4 bracci (20 kDa)</td><td>PEGWorks Creativi</td><td>PSB-455</td><td>https://creativepegworks.com/product/4-arm-peg-mal-mw-20k</td></tr><tr><td>Targhe AggreWell 400</td><td>Tecnologie STEMCELL</td><td>34411</td><td>http://stemcell.com/products/aggrewell400.html</td></tr><tr><td>Etalon distanziato nell'aria</td><td>Macchinari Leggeri</td><td>HF-16074-660</td><td>https://lightmachinery.com/spectrometers/brillouin-spectroscopy-excitation-laser-optimization/</td></tr><tr><td>Soluzione antiaderente per risciacquare</td><td>Tecnologie STEMCELL</td><td>7010</td><td>https://www.stemcell.com/products/aggrewell-rinsing-solution.html</td></tr><tr><td>Incubatore Bold Line Top Stage</td><td>Okolab</td><td>H301-NIKON-NZ100/200/500-N</td><td>https://www.oko-lab.com/live-cell-imaging/stage-top-digital-gas/chamber/nikon/h301-nikon-nz100-200-500-n</td></tr><tr><td>Cobolt Flamenco-100 laser 660 nm</td><td>Huebner</td><td>0660-05-01-0100-700</td><td>https://hubner-photonics.com/products/lasers/single-frequency-lasers/05-01-series/</td></tr><tr><td>Circolatore confocale</td><td>Macchinari Leggeri</td><td>HF-13311</td><td>N/A</td></tr><tr><td>Sistema di illuminazione a fluorescenza D-LEDi &nbsp;</td><td>Nikon</td><td>MBF83000</td><td>https://www.microscope.healthcare.nikon.com/en_EU/products/light-sources/d-ledi</td></tr><tr><td>Modulo EPI-FL</td><td>Nikon</td><td>Ti2-LA-FL-3</td><td>https://www.microscope.healthcare.nikon.com/en_EU/products/accessories/intermediate-modules</td></tr><tr><td>Fibronectina (umana)</td><td>YoProteins</td><td>663</td><td>https://www.yoproteins.com/fibronectin-human-5-mg-663.html</td></tr><tr><td>Gibco Dulbecco' s Eagle Medium Modificato (DMEM)</td><td>ThermoFisher</td><td>11965092</td><td>https://www.thermofisher.com/order/catalog/product/11965092</td></tr><tr><td>Spettrometro HyperFine Brillouin</td><td>Macchinari Leggeri</td><td>HF-8999-PK-660</td><td>https://lightmachinery.com/spectrometers/brillouin-hyperfine-spectrometer/</td></tr><tr><td>Cubo filtro quadruplo a bande LED-DA/FI/TR/Cy5-B</td><td>Nikon</td><td>MXR00753</td><td>https://www.microscope.healthcare.nikon.com/en_EU/products/accessories/fluorescent-filter-cubes</td></tr><tr><td>Cellule staminali mesenchimali (MSC)</td><td>PromoCell</td><td>C-12974</td><td>https://promocell.com/us_en/human-mesenchymal-stem-cells-hmsc.html</td></tr><tr><td>Fase di traduzione motorizzata &nbsp;</td><td>Prior Scientific</td><td>H117E2NN</td><td>https://www.prior.com/imaging-components/motorized-stages?filter=MicroscopeBrand-33</td></tr><tr><td>NucBlue e commercio; Pronti e scambi dal vivo; reagente (Hoechst 33342)</td><td>ThermoFisher</td><td>R37605</td><td>https://www.thermofisher.com/order/catalog/product/R37605</td></tr><tr><td>Obiettivo, S Plan Fluor 20X NA 0.45, aria</td><td>Nikon</td><td>MRH08230</td><td>https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1708</td></tr><tr><td>Fotocamera CMOS ORCA-Fusion</td><td>Hamamatsu</td><td>C14440-20UP</td><td>https://www.hamamatsu.com/eu/en/product/cameras/cmos-cameras/C14440-20UP.html</td></tr><tr><td>PEG-ditiolo, 2 kDa</td><td>PEGWorks Creativi</td><td>PLS-613</td><td>https://creativepegworks.com/product/hs-peg-sh-mw-2k-10g</td></tr><tr><td>sCMOS camera&nbsp;</td><td>Soluzioni Teledyne Vision</td><td>01-IRIS-15-USB-M-16-C</td><td>https://www.teledynevisionsolutions.com/en-gb/products/iris/?model=01-IRIS-15-USB-M-16-C&vertical=tvs-photometrics&segment=tvs&nbsp;</td></tr><tr><td>SPY555-actin&nbsp;</td><td>Spirocromo</td><td>CY-SC202</td><td>https://spirochrome.com/product/spy555-actin/</td></tr><tr><td>TemperatureApp</td><td></td><td></td><td>Software per il controllo della temperatura</td></tr><tr><td>Peptide VPM, GCRDVPMSMRGGDRCG</td><td>GenScript</td><td>N/A</td><td>N/A</td></tr><tr><td>Microscopio Eclipse Ti-2A</td><td>Nikon</td><td>Eclipse Ti-2A</td><td>https://www.microscope.healthcare.nikon.com/en_EU/products/inverted-microscopes/eclipse-ti2-series&nbsp;</td></tr></tbody></table>

mappatura meccanica degli sferoidi tramite spettroscopia di brillouin

Mechanical Mapping of Spheroids Using Brillouin Spectroscopy

Mappatura meccanica degli sferoidi tramite spettroscopia di Brillouin

Brillouin spectroscopy, an emerging technique gaining significant interest in biomedical science, allows researchers to gather information related to ...

mechanical-mapping-of-spheroids-using-brillouin-spectroscopy

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Fonte: Huang, Y. et. al., Monitoraggio morfologico e fisiologico longitudinale di sferoidi tumorali tridimensionali mediante tomografia a coerenza ottica. J. Vis. Exp. (2019)Questo video dimostra l'uso della coerenza ottica tomografia (OCT) per monitorare gli sferoidi tumorali in una piastra multipozzetto acquisendo immagini in sezione trasversale ad alta risoluzione. Analizzando i riflessi di luce dalle strutture cellulari, OCT costruisce modelli 3D per valutare caratteristiche morfologiche...

Monitoraggio morfologico e fisiologico degli sferoidi tumorali mediante tomografia a coerenza ottica 

Microrheology-particella di monitoraggio può essere utilizzato per non distruttivo quantificare e spazialmente mappare i cambiamenti nella matrice extracellulare proprietà meccaniche in modelli tumorali 3D.

Misurazione longitudinale di matrice extracellulare rigidità in 3D Tumore modelli utilizzando-particelle inseguimento Microrheology

Qui vi presentiamo un protocollo per costruire un rapido spettrometro Brillouin. Cascading matrice di fase praticamente ripreso (VIPA) etalon raggiungere una velocità di misura più di 1.000 volte più veloce di scansione tradizionale Fabry-Perot spettrometri. Questo miglioramento fornisce i mezzi per l'analisi Brillouin del tessuto e biomateriali a bassi livelli di potenza in vivo.

Alta velocità sub-GHz spettrometro per l&#39;analisi di Brillouin Scattering

Presentiamo un protocollo per l'applicazione della spettroscopia di diffusione della luce Brillouin alle fibre di collagene di tipo I purificate con elastina e tripsina della matrice extracellulare per estrarre le loro proprietà elastiche complete.

Preparazione della matrice extracellulare fibre proteiche per Brillouin Spettroscopia

Descriviamo la costruzione di una rapida spettrometro di continuo-fluttui-stimolato--scattering Brillouin (CW-SBS). Lo spettrometro impiega singola frequenza laser a diodi e un filtro notch-vapor atomico di acquisire spettri di trasmissione dei campioni torbidi/non-torbido con alta risoluzione spettrale a velocità fino a 100 volte più velocemente rispetto a quelli di spettrometri CW-SBS esistenti. Questo miglioramento consente analisi materiale Brillouin ad alta velocità.<br...

Ad alta velocità continuo-fluttui stimolato Brillouin Scattering spettrometro per analisi materiali

Tomografia a coerenza ottica (OCT), una tecnologia di imaging tridimensionale, è stata utilizzata per monitorare e caratterizzare la cinetica di crescita di sferoidi multicellulari del tumore. Precisa quantificazione volumetrica di sferoidi tumore usando un voxel contando approccio e rilevamento di tessuto morto privo di etichetta in sferoidi basato sul contrasto intrinseco attenuazione ottica, sono stati dimostrati.

Longitudinale morfologiche e fisiologiche monitoraggio di sferoidi tridimensionale del tumore usando la tomografia a coerenza ottica

La struttura fotonica della banda consente di comprendere come si propagano le modalità elettromagnetiche confinate all'interno di un cristallo fotonico. Nei cristalli fotonici che incorporano elementi magnetici, tali modalità ottiche confinate e risonanti sono accompagnate da un'attività magneto-ottica migliorata e modificata. Descriviamo una procedura di misurazione per estrarre la struttura a banda magneto-ottica mediante microscopia spaziale di Fourier.

Caratterizzazione magneto-ottica spectrale e sistemata con angolo di nanostrutture fotoniche

Il protocollo descrive la generazione di sferoidi ad alto rendimento per la bioprinting utilizzando l'analisi multiparametrica della loro ossigenazione e morte cellulare su un microscopio a fluorescenza standard. Questo approccio può essere applicato per controllare la vitalità degli sferoidi ed eseguire la standardizzazione, che è importante nella modellazione del tessuto 3D, del microambiente tumorale e della biofabbricazione (micro)tissutale di successo.

Biofabbricazione assistita da microscopia dell'ossigeno a prezzi accessibili di sferoidi multicellulari

Questo protocollo è stato sviluppato per monitorare longitudinalmente le proprietà meccaniche del tessuto della placca neurale durante la neurulazione dell'embrione di pulcino. Si basa sull'integrazione di un microscopio Brillouin e di un sistema di incubazione sul palco, che consente l'imaging meccanico dal vivo del tessuto della placca neurale in embrioni di pulcino in coltura exovo .

Monitoraggio dell'evoluzione meccanica del tessuto durante la chiusura del tubo neurale dell'embrione di pulcino

Spettroscopia Raman è una tecnica adatta per il non-contatto, label-free analisi di cellule viventi, dell'ingegneria tissutale costrutti e tessuti nativi. Fonte specifici per le impronte digitali spettrali possono essere generati ed analizzati utilizzando l'analisi multivariata.

Senza contatto, Label-free monitoraggio delle cellule e matrice extracellulare mediante Spettroscopia Raman

Impara la caratterizzazione meccanica di sferoidi tumorali in matrice 3D tramite micro-spettroscopia Brillouin. Questo metodo non invasivo consente di studiare la meccanobiologia e la progressione patologica.

Mappatura meccanica degli sferoidi tramite spettroscopia di Brillouin

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