Name: Test di protezione amikacin per la quantificazione e visualizzazione dellinvasione delle cellule di Escherichia coli (Video) | JoVE Journal
Uploaded: 2026-07-03T12:40:00.000+00:00
Duration: 9 min 22 s
Description: Scopri il test di protezione amikacin per quantificare e visualizzare l'invasione delle cellule epiteliali da parte di Escherichia coli aderente-invasiva. Questo metodo supera i limiti del saggio tradizionale con gentamicina per isolati antibiotico-resistenti.

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<ol>
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<table><tbody><tr><td>75 cm2 Fiala per coltura tissutale</td><td>Fisher Scientific&nbsp;</td><td>FB012937</td><td></td></tr><tr><td>Amicacina disulfato&nbsp;</td><td>Research Products International&nbsp;</td><td>A20540-25.0</td><td></td></tr><tr><td>Antibiotico-antimicotico (100x)</td><td>Gibco</td><td>15240062</td><td></td></tr><tr><td>Anticorpo anti-E.coli</td><td>Abcam</td><td>ab137967</td><td></td></tr><tr><td>Piastre micropiastre BioLite - 6 pozzetti</td><td>Fisher Scientific&nbsp;</td><td>12-556-004</td><td></td></tr><tr><td>Albumina sierica bovina</td><td>Sigma-Aldrich</td><td>A7906</td><td></td></tr><tr><td>Collageno I, coda di ratto, 100 mg</td><td>Corning</td><td>354236</td><td></td></tr><tr><td>Lamella per conteggio delle cellule Countess</td><td>Invitrogen</td><td>C10312</td><td></td></tr><tr><td>DMEM, alta glucosi</td><td>Gibco</td><td>11965118</td><td></td></tr><tr><td>Siero fetale bovino</td><td>Corning</td><td>35-011-CV</td><td></td></tr><tr><td>Acido acetico glaciale</td><td>Sigma-Aldrich</td><td>A6283</td><td></td></tr><tr><td>Glicina ultrapura</td><td>USBiological</td><td>G8165</td><td></td></tr><tr><td>Anticorpo secondario anti-Coniglio IgG (H + L) adsorbito incrociato, Alexa Fluor 488</td><td>Invitrogen</td><td>A-11008</td><td></td></tr><tr><td>Hoechst 33342, triidrocloruro triidrato</td><td>Invitrogen</td><td>H3570</td><td></td></tr><tr><td>Liofilchem&nbsp;MTS amicacina [AK] 0,016-256 µg/mL</td><td>Fisher Scientific&nbsp;</td><td>22-777-704</td><td>Strisce di test MIC per amicacina</td></tr><tr><td>Soluzione di paraformaldeide, 4% in PBS</td><td>Fisher Scientific&nbsp;</td><td>J19943.K2</td><td></td></tr><tr><td>PBS, 1x</td><td>Corning</td><td>21-040-CV</td><td></td></tr><tr><td>Coprioggetto in vetro tedesco tedesco sterile rivestito con poli-L-lisina per colture cellulari</td><td>Neuvitro</td><td>NC1327375</td><td></td></tr><tr><td>Montante antifade ProLong Gold&nbsp;</td><td>Invitrogen</td><td>P36934</td><td></td></tr><tr><td>Rhodamina-Falloidina</td><td>Invitrogen</td><td>R415</td><td>Rosso-arancione 540/565 nm</td></tr><tr><td>Piastre per colture tissutali trattate in superficie</td><td>Fisher Scientific&nbsp;</td><td>FB012929</td><td>Piastre da 24 pozzetti</td></tr><tr><td>Triton X-100 (Elettroforesi)</td><td>Fisher Scientific&nbsp;</td><td>BP151-100</td><td></td></tr><tr><td>Blu di Tripano</td><td>Invitrogen</td><td>T10282</td><td></td></tr><tr><td>Tripsina-EDTA (0,25%), phenol red</td><td>Gibco</td><td>25200056</td><td></td></tr><tr><td>Tween-20</td><td>Fisher Scientific&nbsp;</td><td>BP337-100</td><td></td></tr></tbody></table>

test di protezione amikacin per la quantificazione e visualizzazione dell'invasione delle cellule di escherichia coli

Amikacin Protection Assay for Quantification and Visualization of Escherichia coli Cell Invasion

Test di protezione amikacin per la quantificazione e visualizzazione dell'invasione delle cellule di Escherichia coli

Epithelial cell invasion is a key phenotype for many enteric pathogens, including Salmonella, Shigella, and Vibrio. An emerging invasive pathogen is ...

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JoVE Journal

Immunology and Infection

Immunologia e Infezioni

Questo video dimostra come misurare l'invasione delle cellule attraverso inserti in coltura cellulare utilizzando una metodologia basata fluorescenza.

Un In vitro FluoroBlok Tumore Assay Invasion

Un protocollo generale per lo studio di invasione delle cellule ospiti da un agente patogeno batterico, concentrandosi...

Invasione di cellule umane da un agente patogeno batterico

Questo protocollo è un test semplice adesione batterica che consiste nel contare il numero di unità formanti colonie batteriche che vengano rispettate su cellule in coltura. Il test è robusto, indipendente dal adesina studiato, e numerose varianti vengono utilizzate nella maggior parte dei laboratori che lavorano su patogenesi batterica.

In vitro di Adesione batterica su mammiferi cellule epiteliali

Modelli di invasione delle cellule tumorali in tre dimensioni della matrice extracellulare meglio riflettere...

Modellazione e imaging 3-Dimensional Invasion cella Collettiva

Fonte: Cayron, J. e Lesterlin, C. Analisi multiscala della crescita batterica sotto stress. J. Vis. Exp. (2019)Questo video mostra la procedura per valutare la filamentazione di E. coli e la replicazione del DNA in un trattamento antibiotico che inibisce la divisione cellulare utilizzando la placcatura per misurare la vitalità e la citometria a flusso per analizzare le dimensioni delle cellule e il contenuto di DNA.

Valutazione della filamentazione indotta da antibiotici in E. coli mediante saggio di placcatura e citometria a flusso

Questo video mostra la microscopia time-lapse quantitativa per valutare la disseminazione di Listeria monocytogenes dipendente dalla rigidità della matrice extracellulare attraverso le cellule endoteliali. Un idrogel più morbido facilita una più rapida diffusione batterica attraverso le cellule endoteliali rispetto alle controparti più rigide, evidenziando l'influenza della rigidità della matrice sulla disseminazione batterica.

Microscopia quantitativa time-lapse per la valutazione della disseminazione batterica dipendente dalla rigidità della matrice extracellulare

Si descrive un metodo per l'inseguimento della rottura endomembrane suscitato dai batteri...

Misure singole cellule di Rottura Vacuolar causate da patogeni intracellulari

Un protocollo dettagliato di un modello murino per enteroemorragica e. coli (enteroemorragico EHEC) colonizzazione tramite bioluminescenza-identificati batteri è presentato. La rilevazione di questi batteri bioluminescenti di un non-invasiva in vivo imaging sistema animali vivi può avanzare la nostra comprensione corrente della colonizzazione di EHEC.

Rilevamento della colonizzazione di Escherichia Coli enteroemorragica in murino Host di sistema Non invasivo In Vivo bioluminescenza

Abbiamo sviluppato un test multi-pozzetto formato poliacrilammide per sondare l'effetto di rigidità di matrice extracellulare sull'infezione batterica delle cellule aderenti. Questo test è compatibile con citometria a flusso, immunostaining e microscopia di forza di trazione, permettendo per la misurazione quantitativa delle interazioni biomeccaniche tra cellule, loro matrice extracellulare e batteri patogeni.

Un'analisi di poliacrilammide-basata di formato multi-pozzetto per studiare l'effetto della matrice extracellulare rigidità sull'infezione batterica delle cellule aderenti

L'attuale protocollo descrive un metodo integrato, indagando la migrazione delle cellule tumorali e l'invasione su un'unica piattaforma in tempo reale, fornendo un'opzione facilmente riproducibile e tempo efficiente per studiare la morfologia e la mobilità delle cellule.

Un flusso di lavoro semplice migrazione/invasione utilizzando un Imager di vivere-cella automatizzata

Questo manoscritto descrive un metodo per utilizzare la microscopia a fluorescenza a singola cellula per visualizzare e quantificare la competizione batterica in cocoltura.

Quantificazione della competizione interbatterica utilizzando l'imaging a fluorescenza a singola cellula

Scopri il test di protezione amikacin per quantificare e visualizzare l'invasione delle cellule epiteliali da parte di Escherichia coli aderente-invasiva. Questo metodo supera i limiti del saggio tradizionale con gentamicina per isolati antibiotico-resistenti.

Test di protezione amikacin per la quantificazione e visualizzazione dell'invasione delle cellule di Escherichia coli

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