Team:Aachen/Project/2D Biosensor
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=== Medium === | === Medium === | ||
- | Prior to using our own device for detection of fluorescence emitted by the sensor chips we used equipment readily available in the lab. A Molecular Imager® Gel Doc<sup>TM</sup> XR+ from BIO-RAD was available which uses UV and white light illuminators. However, only two different filters were available for the excitation ligth wavelength, which resulted in very limitted possibilities for the excitation of fluorescent molecules. For example, it was possible to detect the expression of iLOV in our sensor chips but the detection of GFP was not possible. It was thus determined that the Gel Doc<sup>TM</sup> was not suitable for our project. | + | Prior to using our own device for detection of fluorescence emitted by the sensor chips we used equipment readily available in the lab. A Molecular Imager® Gel Doc<sup>TM</sup> XR+ from BIO-RAD was available which uses UV and white light illuminators. However, only two different filters were available for the excitation ligth wavelength, which resulted in very limitted possibilities for the excitation of fluorescent molecules. For example, it was possible to detect the expression of iLOV in our sensor chips but the detection of GFP was not possible. It was thus determined that the '''Gel Doc<sup>TM</sup> was not suitable for our project'''. |
{{Team:Aachen/Figure|Aachen_ILOV_GFP_HM_1,5h.png|title=iLOV and GFP in the Gel Doc<sup>TM</sup>|subtitle=Sensor cells producing iLOV (A) and GFP (B) 1.5 h after induction.|width=900px}} | {{Team:Aachen/Figure|Aachen_ILOV_GFP_HM_1,5h.png|title=iLOV and GFP in the Gel Doc<sup>TM</sup>|subtitle=Sensor cells producing iLOV (A) and GFP (B) 1.5 h after induction.|width=900px}} | ||
- | Regarding the medium used for our sensor chips, LB medium showed a high background fluorescence when exposed to UV light in the Gel Doc. | + | Regarding the medium used for our sensor chips, LB medium showed a high background fluorescence when exposed to UV light in the Gel Doc. Surprisingly, the background fluorescence resulting from the LB medium was too high to detect a signal emitted by our sensor cells. Hence, minimal media (NA, M9, Hartman (HM))was used to minimize background fluorescence, but this approachresulted in less to no growth of our sensor cells. |
{{Team:Aachen/Figure|Aachen_Chip_medium_geldoc.png|title=Differend medium in the Gel Doc<sup>TM</sup>|subtitle=Full media (LB) exhibited high background fluorescence while less background fluorescence was observed with the minimal media (HM, M9, NA).|width=900px}} | {{Team:Aachen/Figure|Aachen_Chip_medium_geldoc.png|title=Differend medium in the Gel Doc<sup>TM</sup>|subtitle=Full media (LB) exhibited high background fluorescence while less background fluorescence was observed with the minimal media (HM, M9, NA).|width=900px}} |
Revision as of 20:24, 16 October 2014
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