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

Forfatterne ønsker at takke support relevant for dette manuskript fra Europa-Kommissionen syvende rammeprogram (PIOF-GA-2009 til 255.466) og den irske Health Research Board (HRA_POR/2010/138). Lux-mærket E. coli var en slags gave fra Dr. Cormac Gahan, University College Cork.

1. Tumorinduktion <ol><li> Til rutinemæssig tumorinduktion blev den minimale tumorigen dosis af celler suspenderet i 200 pi serumfrit kulturmedium injiceret subkutant (sc) i flanken af ​​infektion fri 6-8 uger gamle Balb / C eller athymiske MF1-nu/nu mus n = 6 (Harlan, Oxfordshire, UK) (1 x 10 6 4T1-celler) under anvendelse af en 21-gauge kanyle. Levedygtigheden af ​​celler anvendt til podning var større end 95% som bestemt ved visuel tælling med et hæmocytometer og trypanblåt farveeksklusion (...

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I forbindelse med genterapi er anvendelsen af biologiske midler til afgivelse af terapeutiske gener til patienter viste lovende 3-5. Som virus, tillader den iboende biologiske egenskaber af bakterier effektiv DNA-levering til celler eller væv, især i forbindelse med cancer. Det er blevet vist, at bakterier er naturligt i stand til målsøgning til tumorer, når systemisk administrerede resulterer i høje niveauer af replikation lokalt, enten uden for (ikke-invasive arter) eller inden for tumorceller (patoge...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> 4T1-cellelinie </td><td> ATCC </td><td> CRL-2539 </td><td> Syngenisk brystkræft model afledt fra et spontant opstår BALB / c brysttumor </td></tr><tr><td> DMEM </td><td> Sigma-Aldrich </td><td> D6429 </td><td> Dulbeccos modificerede Eagles medium </td></tr><tr><td> PBS </td><td> Sigma-Aldrich </td><td> D8537 </td><td> Phosphatbufret saltvand </td></tr><tr><td> Xenogen IVIS </td><td> Caliper Life Sciences </td><td></td><td> IVIS 100 for 2D-billedbehandling, IVIS Spectrum til 3D. </td></tr><tr><td> Luria Broth Miller (LB) </td><td> Sigma-Aldrich </td><td> L2542 </td><td> Vækstmedium for E. coli </td></tr&gt; <tr><td> Erythromycin </td><td> Sigma-Aldrich </td><td> E5389 </td><td> Antibiotisk </td></tr><tr><td> Streptomycin </td><td> Sigma-Aldrich </td><td> S9137 </td><td> Antibiotisk </td></tr><tr><td> MF1nu/nu mus </td><td> Harlan (UK) </td><td> 069 (nu) / 070 (nu / +) </td><td> HSD: athymiske nøgne-Foxn1nu </td></tr><tr><td> Balb / c mus </td><td> Harlan (UK) </td><td> 066 </td><td> Haplotype: H-2d </td></tr><tr><td> Gavage nål </td><td> Vet-tech Solutions (UK) </td><td> DE009 </td><td> 22G x 38mm lige sondeernæring nål </td></tr><tr><td> Sprøjte til intravenøs injektion </td><td> BD Biosciences </td><td> 309309-1 ml </td><td> Insulinsprøjte med 28 G x ½ inch mikro-fine IV kanyle. </td></tr><tr><td> Sprøjte til tumorpodning </td><td> Braun </td><td> 9161376V </td><td> Omnifix 26 G x &amp;frac12; inch nål </td></tr></tbody></table>

bioluminescent bacterial imaging in vivo

Denne video beskriver brugen af ​​hele kroppen bioluminesce imaging (BLI) for studiet af bakteriel handel med levende mus, med en vægt på brugen af ​​bakterier i gen-og celleterapi til kræft. Bakterier udgør en attraktiv klasse af vektor til cancerterapi, der besidder en naturlig evne til at vokse fortrinsvis i tumorer efter systemisk indgivelse. Bakterier manipuleret til at udtrykke lux genkassette tillader BLI påvisning af bakterier og samtidigt tumorsteder. Placeringen og indholdet af bakterier i tumorer med tiden kan let undersøges, visualiseret i to eller tre dimensioner. Fremgangsmåden kan anvendes på en lang række bakteriearter og tumor xenograft typer. Denne artikel beskriver protokol for analyse af bioluminescerende bakterier i subkutane tumorbærende mus. Visualisering af kommensale bakterier i mave-tarmkanalen (GIT) af BLI er også beskrevet. Denne kraftfulde, og billig, real-time scanning strategi repræsents en ideel metode til undersøgelse af bakterier in vivo i forbindelse med cancerforskning, navnlig genterapi, og infektionssygdomme. Denne video beskriver proceduren for at studere lux-mærket E. coli i levende mus, hvilket viser den rumlige og tidsmæssige udlæsning opnåelige udnytte BLI med IVIS systemet.

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

Bioluminescent Bacterial Imaging In Vivo

Denne artikel beskriver administrationen af<em&gt; Lux-tagget</em&gt; Bakterier til mus og efterfølgende<em&gt; In vivo</em&gt; Analyse under anvendelse IVIS bioluminescens billeddannelse.

Bioluminescerende Bakteriel 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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Immunology and Infection

Bioluminescent Bacterial Imaging In Vivo

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