Name: Imaging 3D ad alta risoluzione e analisi con apprendimento automatico senza marcatura di organoidi intestinali tramite olotomografia a bassa coerenza (Video) | JoVE Journal
Uploaded: 2025-08-12T13:15:00.000+00:00
Duration: 10 min 40 s
Description: Scopri l'immagine 3D ad alta risoluzione di organoidi intestinali senza marcatura tramite olotomografia a bassa coerenza. Questo metodo computazionale consente l'analisi quantitativa guidata dall'intelligenza artificiale per studiare le risposte ai farmaci in tempo reale.

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</ol>

<table><tbody><tr><td>Piastra a 48 pozzetti&nbsp;</td><td>Corning</td><td>3548</td><td>Piatto di coltura</td></tr><tr><td>Soluzione per il recupero cellulare&nbsp;</td><td>Corning</td><td>354253</td><td>Degradazione del Matrigel&nbsp;</td></tr><tr><td>Cisplatino</td><td>Supelco</td><td>PHR1624</td><td>N/A</td></tr><tr><td>Dimetilsolfossido (DMSO)</td><td>Sigma-Aldrich</td><td>D2650</td><td>Veicolo</td></tr><tr><td>Tubo di Eppendorf</td><td>Corning</td><td>MCT-175-C-S</td><td>N/A</td></tr><tr><td>Server di elaborazione HTX</td><td>Tomocube Inc., Corea</td><td>N/A</td><td>Server di elaborazione dati</td></tr><tr><td>HT-X1</td><td>Tomocube Inc., Corea</td><td>N/A</td><td>Software operativo</td></tr><tr><td>Ilastik</td><td>N/A</td><td>N/A</td><td>Toolkit di segmentazione basato sull'apprendimento automatico</td></tr><tr><td>Porta vaso per piatto di imaging&nbsp;</td><td>Tomocube Inc., Corea</td><td>N/A</td><td>Supporto per piastra di imaging</td></tr><tr><td>IntestiCult Terreno di Crescita Organoidi (Topo)</td><td>Tecnologie STEMCELL</td><td>#06005</td><td>Terreno di coltura prefabbricato disponibile in commercio&nbsp;</td></tr><tr><td>Matlab</td><td>MathWorks</td><td>Matlab 2024a</td><td>Software per l'elaborazione delle immagini</td></tr><tr><td>Matrigel</td><td>Corning</td><td>356231</td><td>ECM per organoidi 3D</td></tr><tr><td>Milliporo&nbsp; Steritop&nbsp; Filtro per bottiglie sottovuoto</td><td>Merck</td><td>S2GPT05RE</td><td>0,22 &mu; Unità filtrante M per sterilizzazione media</td></tr><tr><td>Organoidi Intestinali di Topo</td><td>Tecnologie STEMCELL</td><td>#70931</td><td>N/A</td></tr><tr><td>Rifrattometro</td><td>ATAGO</td><td>R-5000</td><td>Calcolatore dell'indice di rifrazione</td></tr><tr><td>TomoDish</td><td>Tomocube Inc., Corea</td><td>901002-02</td><td>#1.5 Piatto di imaging con fondo coprioggetto</td></tr><tr><td>TomoStudio X</td><td>Tomocube Inc., Corea</td><td>N/A</td><td>Software di imaging</td></tr><tr><td>TrypLE</td><td>Gibco</td><td>12604</td><td>Dissociazione di singole cellule</td></tr></tbody></table>

imaging 3d ad alta risoluzione e analisi con apprendimento automatico senza marcatura di organoidi intestinali tramite olotomografia a bassa coerenza

Label-free, High-Resolution 3D Imaging and Machine Learning Analysis of Intestinal Organoids via Low-Coherence Holotomography

La Figura 1 illustra il flusso di lavoro schematico per l'imaging 4D e l'analisi quantitativa delle sIO. Questo protocollo consente l'imaging continuo dal vivo e la valutazione completa delle sIO attraverso un processo in quattro fasi: coltura, imaging 4D, elaborazione delle immagini e analisi quantitativa (Figura 1D).
Dopo un periodo di incubazione di 5 giorni, le sIO mature sono state trasferite in piastre di imaging con fondo copri...

Imaging 3D ad alta risoluzione e analisi con apprendimento automatico senza marcatura di organoidi intestinali tramite olotomografia a bassa coerenza

Accurate, label-free imaging of intestinal organoids is crucial for studying their morphology, growth dynamics, and responses to environmental stimuli. ...

label-free-high-resolution-3d-imaging-machine-learning-analysis

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Biologia

Lensfree ottico tomografia è una tecnica tridimensionale microscopia che offre una risoluzione spaziale di &lt;1 um x &lt;1 um x &lt;3 pm di x, y e Z, rispettivamente, per un grande volume di imaging di 15-100...

Lensfree On-chip Tomographic Microscopia Utilizzando Multi-angolo di illuminazione e Pixel Super-risoluzione

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

L'intera struttura 3D e il contenuto cellulare degli organoidi, così come la loro somiglianza fenotipica con il tessuto originale possono essere catturati utilizzando il protocollo di imaging 3D a risoluzione singola descritto qui. Questo protocollo può essere applicato a una vasta gamma di organoidi che variano in origine, dimensione e forma.

Risoluzione a una cellula di immagini tridimensionali di organoidi intatti

Descriviamo la preparazione di nastri di sezioni seriali e la loro raccolta su un ampio supporto di trasferimento per l'uso come campioni di tomografia array, insieme a procedure di imaging automatizzate in un microscopio elettronico a scansione. Il protocollo consente lo screening, il recupero e l'imaging mirato di eventi locali rari e l'acquisizione di grandi volumi di dati.

Flusso di lavoro di tomografia ad array per l'acquisizione mirata di informazioni sul volume utilizzando la microscopia elettronica a scansione

Qui, forniamo protocolli dettagliati, robusti e complementari per eseguire la colorazione e l'imaging a risoluzione subcellulare di modelli di coltura cellulare tridimensionale fissi che vanno da 100 μm a diversi millimetri, consentendo così la visualizzazione della loro morfologia, composizione del tipo di cellula e interazioni.

Colorazione e imaging ad alta risoluzione di modelli organoidi e sferoidi tridimensionali

Questo documento presenta un protocollo di fabbricazione per un nuovo tipo di substrato di coltura con centinaia di microcontenitori per mm2, in cui gli organoidi possono essere coltivati e osservati utilizzando la microscopia ad alta risoluzione. Anche i protocolli di semina cellulare e immunocolorazione sono dettagliati.

Una piattaforma ad alto rendimento per la coltura e l'imaging 3D di organoidi

Questo protocollo descrive un metodo semi-automatico per screening di farmaci organoidi a medio-alto rendimento e un software di analisi delle immagini automatizzato indipendente dal microscopio per quantificare e visualizzare risposte multiparametriche a singolo organoide per catturare l'eterogeneità intratumorale.

Screening multiparametrico di farmaci organoidi tumorali mediante imaging a cellule vive a campo ampio per analisi di massa e organoidi singoli

In questo articolo, descriviamo una procedura che consente il rilevamento a livello di organo di batteri patogeni durante l'infezione e la quantificazione delle attività del reporter fluorescente.

Tomografia tridimensionale tridimensionale ad alta risoluzione di infezioni microbiche

Questo protocollo combina la chiarificazione ottica e l'immunomarcatura per l'imaging confocale a volume completo di organoidi retinici a montaggio intero. Preserva la struttura 3D, consentendo la visualizzazione dettagliata dei percorsi neuronali chiave, migliorando lo studio della maturazione retinica, l'organizzazione spaziale e le sue potenziali applicazioni nella modellazione delle malattie e nella medicina personalizzata.

Visualizzazione di organoidi retinici a montaggio intero con risoluzione cellulare

Un metodo basato sulla microscopia confocale Raman è presentato che offre etichetta senza visualizzazione di lignina nelle pareti delle cellule vegetali e il confronto di lignificazione nei diversi tessuti, campioni o specie.

Senza etichetta In situ</emImaging> di lignificazione in pareti delle cellule vegetali

Scopri l'immagine 3D ad alta risoluzione di organoidi intestinali senza marcatura tramite olotomografia a bassa coerenza. Questo metodo computazionale consente l'analisi quantitativa guidata dall'intelligenza artificiale per studiare le risposte ai farmaci in tempo reale.

Imaging 3D ad alta risoluzione e analisi con apprendimento automatico senza marcatura di organoidi intestinali tramite olotomografia a bassa coerenza (Video) | JoVE Journal

Imaging 3D ad alta risoluzione e analisi con apprendimento automatico senza marcatura di organoidi intestinali tramite olotomografia a bassa coerenza

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