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Team:Warsaw/Calendar-Stage1/29 August 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 (<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> | ||
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<br />30-08-2010.043- TOP LIZ | <br />30-08-2010.043- TOP LIZ | ||
<br />30-08-2010.044- G LIZ | <br />30-08-2010.044- G LIZ | ||
| - | <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 /> | ||
<hr /> | <hr /> | ||
Revision as of 20:04, 27 October 2010
30.08.2010 Monday
Note:We improved the way of cell preparation by using lysozyme or triton to reduce the adherence of cells
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 (RFC 59 standard points 6 and 7)
Data analysis with Cell Quest and Microsoft Excel [Cherry]
legend:
E.coli constructs:
G- GFP
J- J23102+Listeriolysin O+Invasin+GFP
A- pAraC+Listeriolysin O+Invasin+GFP
TOP 10- without construct
LIZ- treated with lysozyme
TRIT- treated with triton
X- untreated
bacteria:
30-08-2010.001- TOP 10
30-08-2010.003- G
30-08-2010.004- J
30-08-2010.005- A
Hela cells with bacteria:
30-08-2010.033- X TRIT
30-08-2010.034- A TRIT
30-08-2010.035- J TRIT
30-08-2010.036- A LIZ
30-08-2010.037- J LIZ
30-08-2010.038- without bacteria TRIT
30-08-2010.039- TOP TRIT
30-08-2010.040- G TRIT
30-08-2010.041- X LIZ
30-08-2010.042- without bacteria LIZ
30-08-2010.043- TOP LIZ
30-08-2010.044- G LIZ
cytometer plots:
FSC- Forward Scatter, cell size
SSC- Side Scatter, cell complexity
FL-1- FL-1 sensor, cell fluorescence
Cytometer data from Cell Quest analysis:
Lysozyme treatment:
Triton treatment:
Data from Cell Quest processed with Microsoft Excel:
TOP- Hela cels with regular TOP E.coli
M1,M2,M3,M4- PIN Units (according to RFC 59 standard point 7)
