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Team:Warsaw/Calendar-Stage1/24 July 2010
From 2010.igem.org
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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 different E.coli types. | <br>1. Incubation of Hela cells with different E.coli types. | ||
| - | <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, but without using lysozyme (<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> | ||
<br> | <br> | ||
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<br />23-07-2010.011- with A (3) | <br />23-07-2010.011- with A (3) | ||
<br /> | <br /> | ||
| - | <br /> | + | <br />cytometer plots: |
| + | <br>FSC- Forward Scatter, cell size | ||
| + | <br>SSC- Side Scatter, cell complexity | ||
| + | <br>FL-1- FL-1 sensor, cell fluorescence | ||
| + | <br> | ||
<br /> | <br /> | ||
<hr /> | <hr /> | ||
Revision as of 19:58, 27 October 2010
23.07.2010 Friday
Note: We are not totally satisfied with the results, because of the performance of the cells during the flow cytometry examination, they where adhering to each other.
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 different types of E.coli [Ania P.]
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 different E.coli types.
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, but without using lysozyme (RFC 59 standard points 6 and 7)
Data analysis with Cell Quest and Microsoft Excel [Cherry]
legend:
E.coli TOP10 constructs:
G- GFP
I- Invasin+GFP
J- J23102+Listeriolysin O+Invasin+GFP
A- pAraC+Listeriolysin O+Invasin+GFP
Hela cells with bacteria:
23-07-2010.007- without bacteria
23-07-2010.018- with G (1)
23-07-2010.019- with G (2)
23-07-2010.020- with G (3)
23-07-2010.015- with I (1)
23-07-2010.016- with I (2)
23-07-2010.017- with I (3)
23-07-2010.012- with J (1)
23-07-2010.013- with J (2)
23-07-2010.014- with J (3)
23-07-2010.008- with A (1)
23-07-2010.010- with A (2)
23-07-2010.011- with A (3)
cytometer plots:
FSC- Forward Scatter, cell size
SSC- Side Scatter, cell complexity
FL-1- FL-1 sensor, cell fluorescence
Data processed with Cell Quest:
Data from Cell Quest processed with Microsoft Excel:
legend:
Hela N- without bacteria
M1,M2,M3,M4- PIN Units (according to RFC 59 standard point 7)
SD- starndard deviation
ERR.-errror
more exact data:
