Name: measurement of cellular chemotaxis with ecis/taxis
Uploaded: 2012-04-01T11:00:00
Description: Watch this Scientific Journal Video about Measurement of Cellular Chemotaxis with ECIS/Taxis at JoVE.com

इस काम के स्वास्थ्य के राष्ट्रीय संस्थान (ES07408 और EB00208) से अनुदान द्वारा समर्थित किया गया.

1. / ECIS टैक्सी इलेक्ट्रोड तैयार <ol><li> इलेक्ट्रोड / ECIS टैक्सियों सरणी (स्लाइड प्रति 8 कक्षों से मिलकर) की सोने की सतह पहले कमरे के तापमान पर 15 मिनट के लिए बाँझ शर्तों के तहत बाँझ विआयनीकृत पानी (2 DH का.) में 10 मि?...

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परख / ECIS टैक्सी के उपन्यास विशेषताओं इसकी वास्तविक समय डेटा के संग्रह को स्वचालित रूप कोशिकाओं chemoattractant का जवाब करने की क्षमता शामिल हैं. हालांकि इस तकनीक का सबसे आम आवेदन व्यक्ति chemotactic gradients सेलुलर प्रतिक्र?...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूची संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> ECIS Zθ </td><td> एप्लाइड बायोफिज़िक्स </td><td></td><td> <a href="http://www.biophysics.com/prodducts_Ecisz0.php">http://www.biophysics.com/prodducts_Ecisz0.php</a> </td></tr><tr><td> ECIS इलेक्ट्रोड ऐरे </td><td> एप्लाइड बायोफिज़िक्स </td><td> 8W chemotaxis </td><td> <a href="http://www.biophysics.com/cultureware.php">http://www.biophysics.com/cultureware.php</a> </td></tr><tr><td> Seakem GTG agarose </td><td> BioWhittaker आण्विक अनुप्रयोगों </td><td> ५०,०७० </td><td></td></tr><tr><td> RPMI1640 </td><td> Cellgro </td><td> 10-040 </td><td></td></tr><tr><td> HyClone भ्रूण गोजातीय सीरम </td><td> थर्मो वैज्ञानिक </td><td> SH300703 </td><td></td></tr><tr><td> पेnicillin स्ट्रेप्टोमाइसिन / </td><td> सांसद Biomedicals </td><td> 1670049 </td><td> पेनिसिलिन में आइयू ५,००० / मिलीलीटर, स्ट्रेप्टोमाइसिन 5 मिलीग्राम / एमएल </td></tr><tr><td> HEPES बफर </td><td> सांसद Biomedicals </td><td> 1688449 </td><td> 1M समाधान, सेल संस्कृति ग्रेड </td></tr><tr><td> 14 गेज स्टेनलेस स्टील (2) 4 इंच प्रवेशनी </td><td> सामान्य प्रयोगशाला आपूर्ति </td><td> 5-8365-1 </td><td> कुंद बिंदु </td></tr></tbody></table>

measurement of cellular chemotaxis with ecis/taxis

Cellular movement in response to external stimuli is fundamental to many cellular processes including wound healing, inflammation and the response to infection. A common method to measure chemotaxis is the Boyden chamber assay, in which cells and chemoattractant are separated by a porous membrane. As cells migrate through the membrane toward the chemoattractant, they adhere to the underside of the membrane, or fall into the underlying media, and are subsequently stained and visually counted 1. In this method, cells are exposed to a steep and transient chemoattractant gradient, which is thought to be a poor representation of gradients found in tissues 2.
Another assay system, the under-agarose chemotaxis assay, 3, 4 measures cell movement across a solid substrate in a thin aqueous film that forms under the agarose layer. The gradient that develops in the agarose is shallow and is thought to be an appropriate representation of naturally occurring gradients. Chemotaxis can be evaluated by microscopic imaging of the distance traveled. Both the Boyden chamber assay and the under-agarose assay are usually configured as endpoint assays.
The automated ECIS/Taxis system combines the under-agarose approach with Electric Cell-substrate Impedance Sensing (ECIS) 5, 6. In this assay, target electrodes are located in each of 8 chambers. A large counter-electrode runs through each of the 8 chambers (Figure 2). Each chamber is filled with agarose and two small wells are the cut in the agarose on either side of the target electrode. One well is filled with the test cell population, while the other holds the sources of diffusing chemoattractant (Figure 3). Current passed through the system can be used to determine the change in resistance that occurs as cells pass over the target electrode. Cells on the target electrode increase the resistance of the system 6. In addition, rapid fluctuations in the resistance represent changes in the interactions of cells with the electrode surface and are indicative of ongoing cellular shape changes. The ECIS/Taxis system can measure movement of the cell population in real-time over extended periods of time, but is also sensitive enough to detect the arrival of a single cell at the target electrode. 
Dictyostelium discoidium is known to migrate in the presence of a folate gradient 7, 8 and its chemotactic response can be accurately measured by ECIS/Taxis 9. Leukocyte chemotaxis, in response to SDF1&alpha; and to chemotaxis antagonists has also been measured with ECIS/Taxis 10, 11. An example of the leukocyte response to SDF1&alpha; is shown in Figure 1.

Watch this Scientific Journal Video about Measurement of Cellular Chemotaxis with ECIS/Taxis at JoVE.com

Measurement of Cellular Chemotaxis with ECIS/Taxis

ECIS / टैक्सी प्रणाली एक स्वचालित वास्तविक समय परख है कि सेलुलर chemotaxis उपाय है. इस परख में, कोशिकाओं में agarose की एक परत करने के लिए एक लक्ष्य इलेक्ट्रोड पर पहुंचने के नीचे चलते हैं. सेलुलर आंदोलन एसी चालू 0 से प्रतिरोध की शुरुआत द्वारा मापा जाता है.

ECIS / टैक्सी के साथ सेलुलर chemotaxis का मापन

Cellular movement in response to external stimuli is fundamental to many cellular processes including wound healing, inflammation and the response to ...

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