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!align="center"|[[Team:Penn|Home]]
 
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!align="center"|[[Team:Penn/Team|Team]]
 
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!align="center"|[https://igem.org/Team.cgi?year=2010&team_name=Penn Official Team Profile]
 
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!align="center"|[[Team:Penn/Project|Project]]
 
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!align="center"|[[Team:Penn/Parts|Parts Submitted to the Registry]]
 
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!align="center"|[[Team:Penn/Modeling|Modeling]]
 
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!align="center"|[[Team:Penn/Notebook|Notebook]]
 
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!align="center"|[[Team:Penn/Safety|Safety]]
 
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!align="center"|[[Team:Penn/Attributions|Attributions]]
 
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</head>
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==Notebook==
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<body class="home blog logged-in admin-bar chrome">
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You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.
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<h2 class="post-title">Notebook</h2>
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<br />
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<h4>09/20/2011 (Avin, Mike M, Peter, Anthony)</h4>
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<p>First time doing calcium imaging. Brought over 293T's in a 6 well plate, which had been transfected with CatCh. Also had positive control cells on which we used ionophores. Used the Meaney Lab confocal and x-rhod1 dye. Media had 2mM Calcium.
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1uM Ionomycin addition did not change x-rhod1 fluorescent intensity in Anthony's neurons.
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<br /><br />
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Also did not work on our 293T's, but we had the problem of being unable to cleanly pipet the ionomycin into the 6 well plates because the wells we looked at with the scope became entirely blocked by the scope lens.
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Illumination of CatCh cells with 476nm laser from confocal did not increase x-rhod1 intensity. eYFP expressing cells were identified, so transfection worked.
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Will try again Thursday and Friday with larger plates, and possibly switch to FURA-2 and Avin's laser if the confocal still doesn't work. Anthony is going to try further with x-rhod1 dye in neurons to make sure that the dye/laser parameters aren't the problem. Will also attempt the co-culture experiment on these dates.
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<p id="copyright">2011 Penn iGEM Team.</p>
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Latest revision as of 02:11, 25 September 2011

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd"> Penn iGEM 2011 |

Notebook


09/20/2011 (Avin, Mike M, Peter, Anthony)

First time doing calcium imaging. Brought over 293T's in a 6 well plate, which had been transfected with CatCh. Also had positive control cells on which we used ionophores. Used the Meaney Lab confocal and x-rhod1 dye. Media had 2mM Calcium. 1uM Ionomycin addition did not change x-rhod1 fluorescent intensity in Anthony's neurons.

Also did not work on our 293T's, but we had the problem of being unable to cleanly pipet the ionomycin into the 6 well plates because the wells we looked at with the scope became entirely blocked by the scope lens. Illumination of CatCh cells with 476nm laser from confocal did not increase x-rhod1 intensity. eYFP expressing cells were identified, so transfection worked. Will try again Thursday and Friday with larger plates, and possibly switch to FURA-2 and Avin's laser if the confocal still doesn't work. Anthony is going to try further with x-rhod1 dye in neurons to make sure that the dye/laser parameters aren't the problem. Will also attempt the co-culture experiment on these dates.

Retrieved from "http://2011.igem.org/Team:Penn/Notebook"