Name: in vivo imaging of transgenic leishmania parasites in a live host
Uploaded: 2010-07-27T17:00:00
Description: Watch this Scientific Journal Video about In vivo Imaging of Transgenic Leishmania Parasites in a Live Host at JoVE.com

Questo lavoro è stato finanziato in parte da una borsa di studio al merito recensione dal Department of Veterans Affairs, da sovvenzioni NIH AI045540, AI067874, AI076233-01 e AI080801 (MEW), e da AI29646 (SMB). Il lavoro è stato svolto in parte durante finanziamento di CT e JG dal NIH T32 AI07511.

1. L&#39;infezione di piccoli animali transgenici con Leishmania 1. Parasite linee Transgenici Leishmania spp. parassiti che esprimono luciferasi vengono generate utilizzando un episomiale o un vettore di integrazione, come riportato. 1 2 linee clonale sono preferiti. Due punti importanti sono: (A) luciferasi integrato è preferibile luciferasi episomiale, dal momento che in teoria queste righe parassita deve c...

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Il sistema di imaging in vivo (IVIS) fornisce un metodo per l&#39;imaging animale intero o in vivo modelli sperimentali di infezione di imaging diverse forme di leishmaniosi. 18,16 La Leishmania spp. parassiti può essere progettato per esprimere la luciferasi lucciola a un livello che viene rilevato in vivo con la tecnologia di imaging IVIS. Uno dei principali vantaggi di questo metodo è che permette non invasivo visualizzazione di Leishmania spp. dentro l&#39;os...

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<td>D-Luciferin Potassium Salt </td>
<td>Reagent</td>
<td>Caliper LifeSciences (Formerly Xenogen)</td>
<td>122796</td>
<td>&nbsp;</td>
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<td>IVIS Imaging System 200 Series</td>
<td>Equipment</td>
<td>Caliper LifeSciences</td>
<td>&nbsp;</td>
<td>Other IVIS models that can be used include: Lumina II, Lumina XR, Kinetic, and Spectrum. </td>
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in vivo imaging of transgenic leishmania parasites in a live host

Specie diverse di<em&gt; Leishmania,</em&gt; Un protozoo parassita della famiglia<em&gt; Trypanosomatidae</em&gt;, In genere causano diverse manifestazioni della malattia umana. Le forme più comuni della malattia sono la leishmaniosi viscerale (VL) e la leishmaniosi cutanea (CL). Modelli murini di leishmaniosi sono ampiamente utilizzati, ma la quantificazione degli oneri parassiti durante la malattia murino richiede topi per l&#39;eutanasia in vari momenti dopo l&#39;infezione. Carichi parassita vengono poi misurate al microscopio, limitando test di diluizione, qPCR o amplificazione del DNA del parassita. Il<em&gt; In vivo</em&gt; Immagine di sistema (IVIS) è un pacchetto software integrato che consente il rilevamento di un segnale bioluminescente associato con le cellule negli organismi viventi. Sia per ridurre al minimo dell&#39;uso di animali e di seguire longitudinalmente infezioni negli individui, nei modelli in vivo per l&#39;imaging<em&gt; Leishmania</em&gt; Spp. causando VL o CL sono stati stabiliti. I parassiti sono stati progettati per esprimere la luciferasi, e questi sono stati introdotti in topi o intradermica o endovenosa. Misurazioni quantitative della bioluminescenza guida luciferasi del transgenico<em&gt; Leishmania</em&gt; Parassiti all&#39;interno del mouse sono stati realizzati utilizzando IVIS. Topi individuali possono essere ripreso più volte durante gli studi longitudinali, che ci permette di valutare la variabilità inter-animale nel inoculi iniziali parassita sperimentali, e di valutare la moltiplicazione dei parassiti nei tessuti del mouse. I parassiti sono rilevati con elevata sensibilità in luoghi cutanea. Anche se è molto probabile che il segnale (fotoni / secondo / parassita) è più basso in organi viscerali profondi della pelle, ma il confronto quantitativo di segnali in siti superficiali rispetto profondo non sono stati fatti. E &#39;possibile che i numeri parassita tra i siti corpo non sono direttamente confrontabili, anche se i carichi parassita nei tessuti stessi possono essere confrontati tra topi. Esempi di una specie visceralizing (<em&gt; L. infantum chagasi</em&gt;) E una specie che causano la leishmaniosi cutanea (<em&gt; L. mexicana</em&gt;) Sono mostrati. La procedura IVIS può essere utilizzata per monitorare e analizzare modelli animali piccoli di una vasta gamma di<em&gt; Leishmania</em&gt; Specie che causano le diverse forme di leishmaniosi umana.

Watch this Scientific Journal Video about In vivo Imaging of Transgenic Leishmania Parasites in a Live Host at JoVE.com

In vivo Imaging of Transgenic Leishmania Parasites in a Live Host

Un<em&gt; In vivo</em&gt; Sistema di imaging viene utilizzato per generare misurazioni quantitative di infezione murino con il protozoo Trypanosomatid<em&gt; Leishmania</em&gt;. Questo è un metodo non invasivo e non letali per il rilevamento di parassiti esprimere luciferasi in molti tessuti nel corso della cronica<em&gt; Leishmania</em&gt; Spp. infezione.

Imaging in vivo del transgenico parassiti Leishmania in un host live

Distinct species of Leishmania, a protozoan parasite of the family Trypanosomatidae, typically cause different human disease manifestations. The most ...

Research

JoVE Journal

Immunology and Infection

In vivo Imaging of Transgenic Leishmania Parasites in a Live Host

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Non invasiva di imaging di homing dei leucociti e la migrazione In vivo</em

Two-photon (2P) microscopy is a high resolution imaging technique that has been broadly adapted by biologists.  The value of 2P microscopy is that it ...

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Leishmania spp. are protozoan parasites that cause leishmaniases, diseases that present a wide spectrum of clinical manifestations from cutaneous to ...

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