Team:ETH Zurich/lab/chip

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==Mold Design and 3D Print Exchange==
==Mold Design and 3D Print Exchange==
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'''All mesh files designed during the project will be made available at the [http://3dprint.nih.gov/ NIH 3D Print Exchange] under the category 'Custom Labware' via our [http://3dprint.nih.gov/users/ethzurichigem2014 ETH_Zurich_iGEM2014] account.
'''All mesh files designed during the project will be made available at the [http://3dprint.nih.gov/ NIH 3D Print Exchange] under the category 'Custom Labware' via our [http://3dprint.nih.gov/users/ethzurichigem2014 ETH_Zurich_iGEM2014] account.
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==3D-Printing and Rapid Prototyping==
==3D-Printing and Rapid Prototyping==
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|'''Figure 5-d''' Printed 96-well plate, all wells (edge length of 5 mm) are connected.
|'''Figure 5-d''' Printed 96-well plate, all wells (edge length of 5 mm) are connected.
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|'''Figure 7-d''' Close up of the final PDMS chip. The channels are well defined and even small structures separated evenly from the mold.
|'''Figure 7-d''' Close up of the final PDMS chip. The channels are well defined and even small structures separated evenly from the mold.
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|{{:Team:ETH_Zurich/Templates/Video|width=300px|id=video2|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/1/18/ETH_Zurich_2014_two_wells_liquid_culture_small.mp4</html>}}
|{{:Team:ETH_Zurich/Templates/Video|width=300px|id=video2|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/1/18/ETH_Zurich_2014_two_wells_liquid_culture_small.mp4</html>}}
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|'''Video 1 The very first diffusion experiment with fluorescent highlighter ink (pyranine).''' The wells of the PDMS chip were filled with LB agar. About 5 μL ink were added in a punched whole on one side of the two wells. ~4500x faster than real-time.
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|'''Video 1 The very first diffusion experiment with fluorescent highlighter ink ([http://en.wikipedia.org/wiki/Pyranine pyranine]).''' The wells of the PDMS chip were filled with [https://2014.igem.org/Team:ETH_Zurich/lab/protocols#LB_medium_from_dehydrated_product LB agar]. About 5 μL ink were added in a punched whole on one side of the two wells. ~4500x faster than real-time.
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|'''Video 2 Diffusion experiment with liquid cultures.''' The wells of the PDMS chip were filled with LB medium, separated by solidified 2% agarose in the channel. The bottom well contained 3OC6-HSL, the top well ''E. coli'' cells with sfGFP under the control of pLux. ~4500x faster than real-time.
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|'''Video 2 Diffusion experiment with liquid cultures.''' The wells of the PDMS chip were filled with [https://2014.igem.org/Team:ETH_Zurich/lab/protocols#LB_medium_from_dehydrated_product LB medium], separated by solidified 2% agarose in the channel. The bottom well contained 3OC6-HSL (~1 mM), the top well ''E. coli'' cells with sfGFP under the control of pLux. ~4500x faster than real-time.
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|{{:Team:ETH_Zurich/Templates/Video|width=300px|id=video3|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/7/7d/ETH_Zurich_2014_AHL_bead_sensor_bead.mp4</html>}}
|{{:Team:ETH_Zurich/Templates/Video|width=300px|id=video3|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/7/7d/ETH_Zurich_2014_AHL_bead_sensor_bead.mp4</html>}}
|{{:Team:ETH_Zurich/Templates/Video|width=300px|id=video4|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/8/8c/ETH_Zurich_2014_sender_receiver_beads_small.mp4</html>}}
|{{:Team:ETH_Zurich/Templates/Video|width=300px|id=video4|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/8/8c/ETH_Zurich_2014_sender_receiver_beads_small.mp4</html>}}
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|'''Video 3  Diffusion experiment with alginate beads in defined liquid medium.''' The wells of the PDMS chip were filled with CB medium. The bottom well contained beads with 3OC6-HSL, the top well ''E. coli'' cells with sfGFP under the control of pLux confined in alginate beads (d=3 mm). ~1850x faster than real-time.
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|'''Video 3  Diffusion experiment with [https://2014.igem.org/Team:ETH_Zurich/lab/bead alginate beads] in defined liquid medium.''' The wells of the PDMS chip were filled with CB medium. The bottom well contained beads with 3OC6-HSL (~1 mM), the top well ''E. coli'' cells with [https://2014.igem.org/Team:ETH_Zurich/lab/sequences#Sensor_Constructs sfGFP under the control of pLux] confined in [https://2014.igem.org/Team:ETH_Zurich/lab/bead alginate beads] (d=3 mm, intially 10<sup>7</sup> cell/beads). ~1850x faster than real-time.
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|'''Video 4 Cell communication experiment with alginate beads in defined liquid medium.''' The wells of the PDMS chip were filled with CB medium. The bottom well contained ''E. coli'' cells expressing LuxI, which catalyzes the production of 3OC6-HSL; the top well contained ''E. coli'' cells with sfGFP under the control of pLux, All cells were confined in alginate beads (d=3 mm). ~3450x faster than real-time.
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|'''Video 4 Cell communication experiment with [https://2014.igem.org/Team:ETH_Zurich/lab/bead alginate beads] in defined liquid medium.''' The wells of the PDMS chip were filled with CB medium. The bottom well contained ''E. coli'' [https://2014.igem.org/Team:ETH_Zurich/lab/sequences#Producer_Constructs cells expressing LuxI], which catalyzes the production of 3OC6-HSL; the top well contained ''E. coli'' cells with [https://2014.igem.org/Team:ETH_Zurich/lab/sequences#Sensor_Constructs sfGFP under the control of pLux]. All cells were confined in [https://2014.igem.org/Team:ETH_Zurich/lab/bead alginate beads] (d=3 mm, intially 10<sup>7</sup> cell/beads). ~3450x faster than real-time.
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|colspan="2"|{{:Team:ETH_Zurich/Templates/Video|width=600px|id=video5|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/a/a9/ETH_Zurich_2014_signal_propagation.mp4</html>}}
|colspan="2"|{{:Team:ETH_Zurich/Templates/Video|width=600px|id=video5|ratio=143/100|srcMP4=<html>https://static.igem.org/mediawiki/2014/a/a9/ETH_Zurich_2014_signal_propagation.mp4</html>}}
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|colspan="2"|'''Video 5 Row wise, self-propagating [https://2014.igem.org/Team:ETH_Zurich/project/background/biotools#Quorum_Sensing cell-to-cell communication] of ''E. coli'' cells confined in [https://2014.igem.org/Team:ETH_Zurich/lab/bead alginate beads] (d=3 mm, intially 10<sup>7</sup> cell/beads) on a [https://2014.igem.org/Team:ETH_Zurich/lab/chip custom-made millifluidic PDMS chip].''' All cells contained [https://2014.igem.org/Team:ETH_Zurich/expresults/rr#Riboregulators riboregulated] sfGFP followed by [http://parts.igem.org/Part:BBa_C0161 LuxI (BBa_C0161)] together under the control of the [http://parts.igem.org/Part:BBa_R0062 pLux promoter (BBa_R0062)], and [http://parts.igem.org/Part:BBa_J23100 constitutively (BBa_J23100)] expressed [http://parts.igem.org/Part:BBa_C0062 LuxR (BBa_C0062)]. LuxI catalyzes the production of the autoinducer 3OC6-HSL, which is then diffusing from cell to cell. For initialization, the cells in one bead of the top row were induced with 3OC6-HSL before encapsulation. 1750x faster than real-time, the video starts 7 h after the initiation of the experiment.
|colspan="2"|'''Video 5 Row wise, self-propagating [https://2014.igem.org/Team:ETH_Zurich/project/background/biotools#Quorum_Sensing cell-to-cell communication] of ''E. coli'' cells confined in [https://2014.igem.org/Team:ETH_Zurich/lab/bead alginate beads] (d=3 mm, intially 10<sup>7</sup> cell/beads) on a [https://2014.igem.org/Team:ETH_Zurich/lab/chip custom-made millifluidic PDMS chip].''' All cells contained [https://2014.igem.org/Team:ETH_Zurich/expresults/rr#Riboregulators riboregulated] sfGFP followed by [http://parts.igem.org/Part:BBa_C0161 LuxI (BBa_C0161)] together under the control of the [http://parts.igem.org/Part:BBa_R0062 pLux promoter (BBa_R0062)], and [http://parts.igem.org/Part:BBa_J23100 constitutively (BBa_J23100)] expressed [http://parts.igem.org/Part:BBa_C0062 LuxR (BBa_C0062)]. LuxI catalyzes the production of the autoinducer 3OC6-HSL, which is then diffusing from cell to cell. For initialization, the cells in one bead of the top row were induced with 3OC6-HSL before encapsulation. 1750x faster than real-time, the video starts 7 h after the initiation of the experiment.
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Latest revision as of 03:27, 18 October 2014

iGEM ETH Zurich 2014