Team:Warsaw/Calendar-Stage1/10 September 2010

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<br> 10.09.2010 Friday
 
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<br><br>Note: We're not satisfied with the results, because of some problems with the timetable of usage of the cytometer
 
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<br><br><div class="note"> Previous 2 days: Set up and culturing Hela cell culture [Kamil]</div>
 
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<br> Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (<a href="http://openwetware.org/wiki/The_BioBricks_Foundation:RFC#BBF_RFC_59:_Quantitative_measurement_of_mamallian_cell_invasion_by_bacteria_using_flow_cytometry">RFC 59 standard</a> point 5)
 
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<br>
 
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<br><div class="note"> Previous day: Set up liquid cultures of E.coli with different constructs [Milena]</div>
 
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<br> Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (<a href="http://openwetware.org/wiki/The_BioBricks_Foundation:RFC#BBF_RFC_59:_Quantitative_measurement_of_mamallian_cell_invasion_by_bacteria_using_flow_cytometry">RFC 59 standard</a> point 5)
 
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<br>
 
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<br><div class="note"> Testing the invasivnes of E.coli in Hela cells [Kamil,Cherry]</div>
 
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<br>1. Incubation of Hela cells with E.coli with different constructs.
 
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<br>2. Preparation of probes with incubated cells and examining them in flow cytometer.
 
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<br>Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (<a href="http://openwetware.org/wiki/The_BioBricks_Foundation:RFC#BBF_RFC_59:_Quantitative_measurement_of_mamallian_cell_invasion_by_bacteria_using_flow_cytometry">RFC 59 standard</a> points 6 and 7)<br>
 
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<br><div class="note"> Data analysis with Cell Quest and Microsoft Excel [Cherry]</div>
 
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<br>
 
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<br /><strong>legend:</strong>
 
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<hr />
 
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<br />E.coli constructs:
 
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<br />G- GFP
 
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<br />I- Invasin+GFP
 
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<br />J- J23102+Listeriolysin O+Invasin+GFP
 
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<br />A- pAraC+Listeriolysin O+Invasin+GFP
 
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<br />
 
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<br /><strong>Hela cells with bacteria:</strong>
 
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<br />ML.009- without bacteriaN
 
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<br />ML.010- TOP 10 (50)
 
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<br />ML.003- G (25)
 
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<br />ML.004- I (25)
 
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<br />ML.008- I (50)
 
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<br />ML.011- G (50)
 
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<br />10-09-2010.001- J (50)
 
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<br />10-09-2010.002- A (50)
 
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<br />10-09-2010.003- J (25)
 
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<br />
 
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<br /><br />cytometer plots:
 
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<br>FSC- Forward Scatter, cell size
 
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<br>SSC- Side Scatter, cell complexity
 
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<br>FL-1- FL-1 sensor, cell fluorescence
 
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<hr />
 
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Data from the flow cytometer processed by Cell Quest:
 
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<br><br><br/>
 
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<img src="http://2010.igem.org/wiki/images/7/71/Bacto.10.09_01.png" width="450px" />
 
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<img src="http://2010.igem.org/wiki/images/5/59/Bacto.10.09_02.png" width="800px" />
 
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<img src="http://2010.igem.org/wiki/images/1/1a/Bacto.10.09_03.png" width="800px" />
 
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Latest revision as of 23:03, 27 October 2010

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