Name: intravital microscopy of the spleen: quantitative analysis of parasite mobility and blood flow
Uploaded: 2012-01-14T14:00:00
Description: Watch this Scientific Journal Video about Intravital Microscopy of the Spleen: Quantitative Analysis of Parasite Mobility and Blood Flow at JoVE.com

हम विशेष रूप से प्रारंभिक प्रशिक्षण के लिए और मलेरिया परजीवी के intravital माइक्रोस्कोपी में सतत इनपुट एस Graewe और वी. Heussler के लिए आभारी हैं, GFP ट्रांसजेनिक परजीवी दान ए बॉश (Confocal यूनिट, CCiT यूबी, IDIBAPS) के लिए जे बर्न्स छवि विश्लेषण और और तकनीकी सहायता के लिए मात्रा का ठहराव पी. Astola में सहायता के लिए. हम आर Tous और वीडियो उत्पादन के लिए मैं Caralt धन्यवाद. म्यूचुअल फंड कातालुन्या की व्यापकता से एक स्नातक फेलोशिप के प्राप्तकर्ता है. पड़ना एक ICREA अनुसंधान प्रोफेसर है. पड़ना की प्रयोगशाला में काम अनुदान समझौता एन के तहत यूरोपीय समुदाय सातवें फ्रेमवर्क कार्यक्रम (FP7/2007-2013) ° 242095, निजी फाउंडेशन CELLEX (Catalonia, स्पेन), और विज्ञान और नवाचार के स्पेनिश मंत्रालय (द्वारा वित्त पोषित है SAF2009 07,760).

इस विधि 7 में रिपोर्ट अनुसंधान में इस्तेमाल किया गया था. 1. हरी फ्लोरोसेंट प्रोटीन (GFP) ट्रांसजेनिक परजीवी के साथ पशु संक्रमण <ol><li> पी. yoelii - GFP 17XL और 17X के ट्रांसजेनिक लाइनों एक ही वैक्टर उत्पन?...

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इस कृंतक मलेरिया मॉडल में तिल्ली के intravital माइक्रोस्कोपी के कार्यान्वयन इस अंग है जो अब तक एक &quot;ब्लैक बॉक्स&quot; तकनीकी कारणों की वजह से माना गया है के माध्यम से परजीवी के गतिशील यात्रा की जांच की संभावना ख?...

<table border="1"><tbody><tr><td> अभिकर्मक के नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> Leica टीसीएस SP5 confocal सूक्ष्मदर्शी </td><td> Leica माइक्रोसिस्टम्स, हीडलबर्ग, जर्मनी </td><td> TCS-SP5 सीरियल नं. 5100000419 </td><td></td></tr><tr><td> Ketamine (Ketolar 50 मिलीग्राम / एमएल) </td><td> फ़िज़र </td><td> 631028 </td><td></td></tr><tr><td> Midazolam 15 मिलीग्राम / 3 मिलीलीटर </td><td> Normon </td><td> 838193 </td><td></td></tr><tr><td> ७०००० मेगावाट Dextran, टेक्सास रेड के लिए संयुग्मित </td><td> आण्विक जांच </td><td> D1830 </td><td></td></tr><tr><td> Fluorescein आइसोथियोसाइनेट, isomer मैं (FITC) </td><td> सिग्मा </td><td> F7250 </td><td></td></tr><tr><td> 33342 Hoechst </td><td> सिग्मा </td><td> H1399 </td><td></td></tr><tr><td> Giemsa दाग </td><td> सिग्मा </td><td> GS1 </td><td> कार्य समाधान आसुत जल में 10% से कम है </td></tr><tr><td> सुपर गोंद 3 लॉकटाइट </td><td> लॉकटाइट </td><td> 9975-0880 </td><td></td></tr></tbody></table>

intravital microscopy of the spleen: quantitative analysis of parasite mobility and blood flow

The advent of intravital microscopy in experimental rodent malaria models has allowed major advances to the knowledge of parasite-host interactions 1,2. Thus, in vivo imaging of malaria parasites during pre-erythrocytic stages have revealed the active entrance of parasites into skin lymph nodes 3, the complete development of the parasite in the skin 4, and the formation of a hepatocyte-derived merosome to assure migration and release of merozoites into the blood stream 5. Moreover, the development of individual parasites in erythrocytes has been recently documented using 4D imaging and challenged our current view on protein export in malaria 6. Thus, intravital imaging has radically changed our view on key events in Plasmodium development. Unfortunately, studies of the dynamic passage of malaria parasites through the spleen, a major lymphoid organ exquisitely adapted to clear infected red blood cells are lacking due to technical constraints.
Using the murine model of malaria Plasmodium yoelii in Balb/c mice, we have implemented intravital imaging of the spleen and reported a differential remodeling of it and adherence of parasitized red blood cells (pRBCs) to barrier cells of fibroblastic origin in the red pulp during infection with the non-lethal parasite line P.yoelii 17X as opposed to infections with the P.yoelii 17XL lethal parasite line 7. To reach these conclusions, a specific methodology using ImageJ free software was developed to enable characterization of the fast three-dimensional movement of single-pRBCs. Results obtained with this protocol allow determining velocity, directionality and residence time of parasites in the spleen, all parameters addressing adherence in vivo. In addition, we report the methodology for blood flow quantification using intravital microscopy and the use of different colouring agents to gain insight into the complex microcirculatory structure of the spleen. 
Ethics statement
All the animal studies were performed at the animal facilities of University of Barcelona in accordance with guidelines and protocols approved by the Ethics Committee for Animal Experimentation of the University of Barcelona CEEA-UB (Protocol No DMAH: 5429). Female Balb/c mice of 6-8 weeks of age were obtained from Charles River Laboratories.

Watch this Scientific Journal Video about Intravital Microscopy of the Spleen: Quantitative Analysis of Parasite Mobility and Blood Flow at JoVE.com

Intravital Microscopy of the Spleen: Quantitative Analysis of Parasite Mobility and Blood Flow

हम तिल्ली के intravital माइक्रोस्कोपी GFP ट्रांसजेनिक मलेरिया परजीवी और इस अंग के भीतर परजीवी गतिशीलता और रक्त प्रवाह की मात्रा का ठहराव का उपयोग प्रदर्शन के लिए विधि दिखा.

प्लीहा के intravital माइक्रोस्कोपी: परजीवी गतिशीलता और रक्त प्रवाह के मात्रात्मक विश्लेषण

The advent of intravital microscopy in experimental rodent malaria models has allowed major advances to the knowledge of parasite-host interactions 1,2. ...

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