Name: Mechanische mapping van sferoïden met behulp van Brillouin-spectroscopie (Video) | JoVE Journal
Uploaded: 2025-12-12T13:10:00.000+00:00
Duration: 8 min 27 s
Description: Leer mechanische mapping van sferoïden met Brillouin-microspectroscopie voor mechanobiologie. Deze niet-invasieve methode karakteriseert celmechanica in 3D zonder de sample te verstoren.

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</ol>

<table><tbody><tr><td>4-arm PEG-Maleimide (20 kDa)</td><td>Creative PEGWorks</td><td>PSB-455</td><td>https://creativepegworks.com/product/4-arm-peg-mal-mw-20k</td></tr><tr><td>AggreWell 400 plates</td><td>STEMCELL Technologies</td><td>34411</td><td>http://stemcell.com/products/aggrewell400.html</td></tr><tr><td>Air-spaced etalon</td><td>LightMachinery</td><td>HF-16074-660</td><td>https://lightmachinery.com/spectrometers/brillouin-spectroscopy-excitation-laser-optimization/</td></tr><tr><td>Anti-adherence rinsing solution</td><td>STEMCELL Technologies</td><td>7010</td><td>https://www.stemcell.com/products/aggrewell-rinsing-solution.html</td></tr><tr><td>Bold Line Top Stage Incubator</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-100 660 nm laser</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>Confocal circulator</td><td>LightMachinery</td><td>HF-13311</td><td>N/A</td></tr><tr><td>D-LEDi Fluorescence illumination system&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>EPI-FL module</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>Fibronectin (human)</td><td>YoProteins</td><td>663</td><td>https://www.yoproteins.com/fibronectin-human-5-mg-663.html</td></tr><tr><td>Gibco Dulbecco&rsquo;s Modified Eagle Medium (DMEM)</td><td>ThermoFisher</td><td>11965092</td><td>https://www.thermofisher.com/order/catalog/product/11965092</td></tr><tr><td>HyperFine Brillouin spectrometer</td><td>LightMachinery</td><td>HF-8999-PK-660</td><td>https://lightmachinery.com/spectrometers/brillouin-hyperfine-spectrometer/</td></tr><tr><td>LED-DA/FI/TR/Cy5-B Quadruple band filter cube</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>Mesenchymal stem cells (MSCs)</td><td>PromoCell</td><td>C-12974</td><td>https://promocell.com/us_en/human-mesenchymal-stem-cells-hmsc.html</td></tr><tr><td>Motorized translation stage&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&trade; Live ReadyProbes&trade; reagent (Hoechst 33342)</td><td>ThermoFisher</td><td>R37605</td><td>https://www.thermofisher.com/order/catalog/product/R37605</td></tr><tr><td>Objective, S Plan Fluor 20X NA 0.45, air</td><td>Nikon</td><td>MRH08230</td><td>https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1708</td></tr><tr><td>ORCA-Fusion CMOS camera</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-dithiol, 2 kDa</td><td>Creative PEGWorks</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>Teledyne Vision Solutions</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&amp;vertical=tvs-photometrics&amp;segment=tvs&nbsp;</td></tr><tr><td>SPY555-actin&nbsp;</td><td>Spirochrome</td><td>CY-SC202</td><td>https://spirochrome.com/product/spy555-actin/</td></tr><tr><td>TemperatureApp</td><td></td><td></td><td>Temperature control software</td></tr><tr><td>VPM peptide, GCRDVPMSMRGGDRCG</td><td>GenScript</td><td>N/A</td><td>N/A</td></tr><tr><td>Eclipse Ti-2A microscope</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>

mechanische mapping van sferoïden met behulp van brillouin-spectroscopie

Mechanical Mapping of Spheroids Using Brillouin Spectroscopy

Mechanische mapping van sferoïden met behulp van Brillouin-spectroscopie

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

Research

JoVE Journal

Bioengineering

Bio-engineering

Source: Huang, Y. et. al., Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography. J. Vis. Exp. (2019)This video demonstrates the use of optical coherence tomography (OCT) to monitor tumor spheroids in a multi-well plate by capturing high-resolution cross-sectional images. By analyzing light reflections from cellular structures, OCT constructs 3D models to assess morphological features like size and shape while also...

Morphological and Physiological Monitoring of Tumor Spheroids Using Optical Coherence Tomography 

Deeltjes volgen microrheology kunnen worden gebruikt om niet-destructieve kwantificeren en veranderingen in extracellulaire matrix mechanische eigenschappen in 3D tumormodellen ruimtelijk in kaart.

Longitudinale Meting van extracellulaire matrix stijfheid in 3D Tumor modellen met-Particle Tracking Microreologie

Hier presenteren we een protocol om een ​​snelle Brillouin spectrometer te bouwen. Cascading vrijwel afgebeeld fase array (VIPA) etalons bereiken van een meetsnelheid meer dan 1000 keer sneller dan de traditionele scannen Fabry-Perot spectrometers. Deze verbetering is het instrument om Brillouin analyse van weefsel en biomaterialen bij lage energieniveaus in vivo.

High Speed ​​Sub-GHz Spectrometer voor Brillouin Scattering Analyse

We presenteren een protocol voor de toepassing van Brillouin lichtverstrooiingsspectroscopie op elastine en trypsine-gezuiverde type I collageenvezels van de extracellulaire matrix om hun volledige elastische eigenschappen te extraheren.

Bereiding van extracellulaire matrix eiwit vezels voor Brillouin Spectroscopie

Beschrijven we de bouw van een snelle continuous-wave-gestimuleerd-Brillouin-verstrooiing (CW-SBS) spectrometer. De spectrometer heeft één-frequentie-diodelasers en een atomaire damp inkeping-filter te verwerven transmissie spectra van troebel/niet-troebel monsters met hoge spectrale resolutie bij snelheden tot 100-fold sneller dan die van bestaande CW-SBS spectrometers. Deze verbetering maakt een snelle Brillouin materiële analyse.<br...

High-Speed Continuous-wave gestimuleerd Brillouin verstrooiing Spectrometer voor materiaal analyse

Optische coherentie tomografie (OCT), een driedimensionale imaging technologie, werd gebruikt om te controleren en te karakteriseren de kinetiek van de groei van meercellige tumor spheroïden. Precieze volumetrische kwantificering van tumor spheroïden met behulp van een voxel tellen aanpak, en label-vrije dood weefsel detectie in de spheroïden op basis van intrinsieke optische demping daarentegen werden gedemonstreerd.

Longitudinale morfologische en fysiologische Monitoring van driedimensionale Tumor spheroïden met behulp van optische coherentie tomografie

De Photonic bandstructuur maakt het mogelijk om te begrijpen hoe beperkt elektromagnetische modi zich verspreiden in een fotonisch kristal. In fotonische kristallen die magnetische elementen bevatten, gaan dergelijke beperkte en resonerende optische modi gepaard met een verbeterde en gewijzigde Magneto-optische activiteit. We beschrijven een meetprocedure om de Magneto-optische bandstructuur uit te pakken door Fourier-ruimte microscopie.

Spectrale en hoek-opgeloste Magneto-optische karakterisatie van fotonische nanostructuren

Het protocol beschrijft high-throughput sferoïde generatie voor bioprinting met behulp van de multi-parametrische analyse van hun oxygenatie en celdood op een standaard fluorescentiemicroscoop. Deze aanpak kan worden toegepast om de levensvatbaarheid van de sferoïden te controleren en standaardisatie uit te voeren, wat belangrijk is bij het modelleren van 3D-weefsel, tumormicro-omgeving en succesvolle (micro)weefselbiofabricage.

Betaalbare zuurstofmicroscopie-geassisteerde biofabricage van meercellige sferoïden

Dit protocol is ontwikkeld om de mechanische eigenschappen van neuraal plaatweefsel tijdens de neurulatie van kuikenembryo's longitudinaal te monitoren. Het is gebaseerd op de integratie van een Brillouin-microscoop en een incubatiesysteem op het podium, waardoor live mechanische beeldvorming van neuraal plaatweefsel in ex ovo gekweekte kuikenembryo's mogelijk wordt.

Monitoring van de mechanische evolutie van weefsel tijdens het sluiten van de neurale buis van kuikenembryo's

Raman spectroscopie is een geschikte techniek voor de non-contact, label-free analyse van levende cellen, weefsel-engineering constructen en cellen. Bronspecifieke spectrale vingerafdrukken worden gegenereerd en geanalyseerd met multivariate analyse.

Non-contact, Label-free monitoring van cellen en extracellulaire matrix met behulp van Raman spectroscopie

Leer mechanische mapping van sferoïden met Brillouin-microspectroscopie voor mechanobiologie. Deze niet-invasieve methode karakteriseert celmechanica in 3D zonder de sample te verstoren.

Mechanische mapping van sferoïden met behulp van Brillouin-spectroscopie

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