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Team:Warsaw/Calendar-Stage1/10 September 2010
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<br><div class="note"> Testing the invasivnes of E.coli in Hela cells [Kamil,Cherry]</div> | <br><div class="note"> Testing the invasivnes of E.coli in Hela cells [Kamil,Cherry]</div> | ||
<br>1. Incubation of Hela cells with E.coli with different constructs. | <br>1. Incubation of Hela cells with E.coli with different constructs. | ||
| - | <br>2. Preparation of | + | <br>2. Preparation of samples with incubated cells and examining them in flow cytometer. |
<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> | <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> | ||
<br><div class="note"> Data analysis with Cell Quest and Microsoft Excel [Cherry]</div> | <br><div class="note"> Data analysis with Cell Quest and Microsoft Excel [Cherry]</div> | ||
Revision as of 20:25, 27 October 2010
10.09.2010 Friday
Note: We're not satisfied with the results, because of some problems with the timetable of usage of the cytometer
Previous 2 days: Set up and culturing Hela cell culture [Kamil]
Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (RFC 59 standard point 5)
Previous day: Set up liquid cultures of E.coli with different constructs [Milena]
Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (RFC 59 standard point 5)
Testing the invasivnes of E.coli in Hela cells [Kamil,Cherry]
1. Incubation of Hela cells with E.coli with different constructs.
2. Preparation of samples with incubated cells and examining them in flow cytometer.
Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (RFC 59 standard points 6 and 7)
Data analysis with Cell Quest and Microsoft Excel [Cherry]
legend:
E.coli constructs:
G- GFP
I- Invasin+GFP
J- J23102+Listeriolysin O+Invasin+GFP
A- pAraC+Listeriolysin O+Invasin+GFP
25,50- different bacteria concentration
Hela cells with bacteria:
ML.009- without bacteria
ML.010- TOP10 without construct(50)
ML.003- G (25)
ML.004- I (25)
ML.008- I (50)
ML.011- G (50)
10-09-2010.001- J (50)
10-09-2010.002- A (50)
10-09-2010.003- J (25)
cytometer plots:
FSC- Forward Scatter, cell size
SSC- Side Scatter, cell complexity
FL-1- FL-1 sensor, cell fluorescence
Data from the flow cytometer processed by Cell Quest:
