Team:Aachen/Project/2D Biosensor
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{{Team:Aachen/FigureFloatRight|Aachen_K131026_HSLdetection_slow.gif|title=Detection of 3-oxo-C{{sub|12}} HSL with K131026|subtitle=0.2 µL of 3-oxo-C{{sub|12}} HSL was placed in the middle of the chip and then incubated at 37°C in ''WatsOn''.|width=480px}} | {{Team:Aachen/FigureFloatRight|Aachen_K131026_HSLdetection_slow.gif|title=Detection of 3-oxo-C{{sub|12}} HSL with K131026|subtitle=0.2 µL of 3-oxo-C{{sub|12}} HSL was placed in the middle of the chip and then incubated at 37°C in ''WatsOn''.|width=480px}} | ||
+ | ===Detecting the 3-oxo-C{{sub|12}}-HSL with K131026 in our Sensor Chips with ''WatsOn''=== | ||
The next step towards the final goal of detecting living cells of ''Pseudomonas aeruginosa'' was to reproduce the detection of 3-oxo-C{{sub|12}}-HSL, which was established in the plate reader, in our own [https://2014.igem.org/Team:Aachen/Project/Measurement_Device ''WatsOn''] device. Therefore, we again used ''E. coli'' BL21(DE3) cells containing [http://parts.igem.org/Part:BBa_K131026 K131026] and induced with 0.2 µL 3-oxo-C{{sub|12}}-HSL in a concentration of 500 µg/mL. The right chip was induced and - as a negative control - the left chip was not induced. Pictures were taken every four minutes (Detection of 3-oxo-C12 HSL with K131026, displayed below). | The next step towards the final goal of detecting living cells of ''Pseudomonas aeruginosa'' was to reproduce the detection of 3-oxo-C{{sub|12}}-HSL, which was established in the plate reader, in our own [https://2014.igem.org/Team:Aachen/Project/Measurement_Device ''WatsOn''] device. Therefore, we again used ''E. coli'' BL21(DE3) cells containing [http://parts.igem.org/Part:BBa_K131026 K131026] and induced with 0.2 µL 3-oxo-C{{sub|12}}-HSL in a concentration of 500 µg/mL. The right chip was induced and - as a negative control - the left chip was not induced. Pictures were taken every four minutes (Detection of 3-oxo-C12 HSL with K131026, displayed below). | ||
<br>The result was a clear replication of the success of the plate reader experiment. The induced chip showed a clear fluorescence response eminating from the center where the induction with HSL took place, thus demonstrating the ability of not only our sensor chip technology, but also our measurement device ''WatsOn to successfully'' detect 3-oxo-C{{sub|12}}-HSL. | <br>The result was a clear replication of the success of the plate reader experiment. The induced chip showed a clear fluorescence response eminating from the center where the induction with HSL took place, thus demonstrating the ability of not only our sensor chip technology, but also our measurement device ''WatsOn to successfully'' detect 3-oxo-C{{sub|12}}-HSL. | ||
{{Team:Aachen/FigureFloat|Aachen_K131026_Pseudomonas_aeruginosa_detection.gif|title=Detection of ''Pseudomonas aeruginosa'' with K131026|subtitle=Direct detection of ''Pseudomonas aeruginosa'' on our sensor chips. Sensor cell used were K131026.|width=480px}} | {{Team:Aachen/FigureFloat|Aachen_K131026_Pseudomonas_aeruginosa_detection.gif|title=Detection of ''Pseudomonas aeruginosa'' with K131026|subtitle=Direct detection of ''Pseudomonas aeruginosa'' on our sensor chips. Sensor cell used were K131026.|width=480px}} | ||
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===Detecting ''Pseudomonas aeruginosa'' with K131026 in our Sensor Chip with ''WatsOn''=== | ===Detecting ''Pseudomonas aeruginosa'' with K131026 in our Sensor Chip with ''WatsOn''=== | ||
After establishing the successful detection of 3-oxo-C{{sub|12}}-HSLs with our sensor chips the next step was to detect living cells of ''Pseudomonas aeruginosa'' with our measurement device ''WatsOn''. Therefore sensor chips based on [http://parts.igem.org/Part:BBa_K131026 K131026] were prepared and the right chip was induced with 0.2 µl of a ''Pseudomonas aeruginosa'' culture while the left chip was not induced (Detection of 3-oxo-C12 HSL with K131026, displayed below). On the induced chip, a clear fluorescence signal was visible in response to ''P. aeruginosa''. The fluorescence signal emerged outward from the induction point and showed a significant difference to the non-induced chip. The results clearly demonstrate the ability of our sensor chip technology and our measurement device ''WatsOn'' to detect ''P. aeruginosa''! | After establishing the successful detection of 3-oxo-C{{sub|12}}-HSLs with our sensor chips the next step was to detect living cells of ''Pseudomonas aeruginosa'' with our measurement device ''WatsOn''. Therefore sensor chips based on [http://parts.igem.org/Part:BBa_K131026 K131026] were prepared and the right chip was induced with 0.2 µl of a ''Pseudomonas aeruginosa'' culture while the left chip was not induced (Detection of 3-oxo-C12 HSL with K131026, displayed below). On the induced chip, a clear fluorescence signal was visible in response to ''P. aeruginosa''. The fluorescence signal emerged outward from the induction point and showed a significant difference to the non-induced chip. The results clearly demonstrate the ability of our sensor chip technology and our measurement device ''WatsOn'' to detect ''P. aeruginosa''! |
Revision as of 01:59, 18 October 2014
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