Name: bioluminescent bacterial imaging in vivo
Uploaded: 2012-11-04T23:00:00
Description: Watch this Scientific Journal Video about Bioluminescent Bacterial Imaging In Vivo at JoVE.com

De auteurs willen ondersteunen die relevant zijn voor dit manuscript erkennen van de Europese Commissie zevende kaderprogramma (PIOF-GA-2009 tot 255.466 duizend) en de Ierse Health Research Board (HRA_POR/2010/138). Lux-gelabelde E. coli was een vriendelijke gift van Dr Cormac Gahan, University College Cork.

1. Tumor Inductie <ol><li> Voor routine tumorinductie werd de minimale dosis van tumorigene cellen gesuspendeerd in 200 ul serumvrij kweekmedium subcutaan (sc) in de flank van infectievrije 6-8 weken oude vrouwelijke Balb / C muizen of athymische MF1-nu/nu n = 6 (Harlan, Oxfordshire, UK) (1 x 10 6 4T1 cellen) met een 21-gauge naald. De levensvatbaarheid van cellen voor inoculatie bedroeg meer dan 95% zoals bepaald door visuele telling met een hemocytometer en Trypan Blue Dye Exclusion (Gibco). </li><li> Na ...

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In de context van gentherapie, is het gebruik van biologische agentia voor de levering van therapeutische genen aan patiënten grote belofte 3-5. Zoals virussen, de aangeboren biologische eigenschappen van bacteriën mogelijk efficiënte DNA afgifte aan cellen of weefsels, met name in de context van kanker. Aangetoond is dat bacteriën van nature kunnen homing tumoren bij systemische toediening resulteert in hoge niveaus van replicatie lokaal of buiten (niet-invasieve soorten) of binnen tumorcellen (pathogene...

<table border="1"><tbody><tr><td> Naam van het reagens </td><td> Vennootschap </td><td> Catalogusnummer </td><td> Reacties </td></tr><tr><td> 4T1 cellijn </td><td> ATCC </td><td> CRL-2539 </td><td> Syngene borstkankermodel afgeleid van een spontaan ontstaan ​​BALB / c borsttumor </td></tr><tr><td> DMEM </td><td> Sigma-Aldrich </td><td> D6429 </td><td> Dulbecco&#39;s Modified Eagle&#39;s Medium </td></tr><tr><td> PBS </td><td> Sigma-Aldrich </td><td> D8537 </td><td> Fosfaat gebufferde zoutoplossing </td></tr><tr><td> Xenogen IVIS </td><td> Caliper Life Sciences </td><td></td><td> IVIS 100 voor 2D-beeldvorming; IVIS Spectrum voor 3D. </td></tr><tr><td> Luria Broth Miller (LB) </td><td> Sigma-Aldrich </td><td> L2542 </td><td> Groeimedium voor E. coli </td></tr&gt; <tr><td> Erythromycine </td><td> Sigma-Aldrich </td><td> E5389 </td><td> Antibioticum </td></tr><tr><td> Streptomycine </td><td> Sigma-Aldrich </td><td> S9137 </td><td> Antibioticum </td></tr><tr><td> MF1nu/nu muizen </td><td> Harlan (UK) </td><td> 069 (nu) / 070 (nu / +) </td><td> HSD: Athymische Naakt-Foxn1nu </td></tr><tr><td> Balb / c muizen </td><td> Harlan (UK) </td><td> 066 </td><td> Haplotype: H-2 d </td></tr><tr><td> Gavage naald </td><td> Vet-tech Solutions (UK) </td><td> DE009 </td><td> 22G x 38mm rechte sonde naald </td></tr><tr><td> Spuit voor intraveneuze injectie </td><td> BD BioSciences </td><td> 309309-1 ml </td><td> Insuline spuit met 28 G x ½ inch micro-fijne IV naald. </td></tr><tr><td> Injectiespuit tumorinoculatie </td><td> Braun </td><td> 9161376V </td><td> Omnifix 26 G x &amp;frac12; inch naald </td></tr></tbody></table>

bioluminescent bacterial imaging in vivo

Deze video beschrijft het gebruik van het gehele lichaam bioluminesce imaging (BLI) voor de studie van bacteriële handel in levende muizen, met een nadruk op het gebruik van bacteriën in gen-en celtherapie voor kanker. Aanwezige bacteriën een aantrekkelijke klasse van vector voor de behandeling van kanker, het bezit van een natuurlijke vermogen om bij voorkeur binnen tumoren groeien na systemische toediening. Bacteriën ontworpen om het lux-gen cassette vergunning BLI detectie van de bacteriën en tegelijkertijd tumorplaatsen drukken. De locatie en de hoeveelheid bacteriën in tumoren tijd gemakkelijk worden onderzocht, gevisualiseerd in twee of drie dimensies. De methode is toepasbaar op een breed scala aan bacteriesoorten en tumor xenograft types. Dit artikel beschrijft het protocol voor analyse van bioluminescente bacteriën in subcutane tumor dragende muizen. Visualisatie van commensale bacteriën in het maag-darmkanaal (GIT) door BLI wordt ook beschreven. Deze krachtige, en goedkoop, real-time beeldvorming strategie vertegents een ideale methode voor de studie van bacteriën in vivo in het kader van kankeronderzoek, met name gentherapie en infectieziekten. Deze video beschrijft de procedure voor het bestuderen van lux-gelabelde E. coli in levende muizen, waaruit de ruimtelijke en temporele uitlezing haalbaar gebruik te maken van BLI met de IVIS-systeem.

Watch this Scientific Journal Video about Bioluminescent Bacterial Imaging In Vivo at JoVE.com

Bioluminescent Bacterial Imaging In Vivo

Dit artikel beschrijft de toediening van<em&gt; Lux-gelabelde</em&gt; Bacteriën muizen en daaropvolgende<em&gt; In vivo</em&gt; Analyse met behulp van bioluminescentie IVIS beeldvorming.

Bioluminescent Bacteriële Imaging<em&gt; In Vivo</em

This video describes the use of whole body bioluminesce imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of ...

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Bioluminescent Bacterial Imaging In Vivo

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