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| | <noinclude>{{:Team:EPF-Lausanne/Template/Header}}</noinclude> | | <noinclude>{{:Team:EPF-Lausanne/Template/Header}}</noinclude> |
| - | {{:Team:EPF-Lausanne/Template/ProtocolHeader|Fluorescence Check|{{{1|}}}}} | + | |
| | + | <noinclude>{{:Team:EPF-Lausanne/Template/SetTitle|Guava Fluorescence Measurement}}</noinclude> |
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| | + | {{:Team:EPF-Lausanne/Template/ProtocolHeader|Fluorescence (Guava)|{{{1|}}}}} |
| | <!-- /\ --> | | <!-- /\ --> |
| | <!-- || --> | | <!-- || --> |
| | <!-- Insert content here (don't forget to set the title up here!) --> | | <!-- Insert content here (don't forget to set the title up here!) --> |
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| | + | Prepare your samples by measuring their PCV (or estimating the cell amount according to the doubling rate). Dilute them with PBS in order to have between 200 and 500 cells/µl. Prepare at least one well that has seed cells. |
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| - | 1. Trash out the waste on the right bottom of the machine before you start. | + | Steps 1 to 4 are optional and should be done from time to time. |
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| - | 2. Add bleach (detergent) to the tubes on the right positions. | + | 1. Trash the waste on the bottom right of the machine if it is full before you start. |
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| | + | 2. Put tubes with bleach (detergent) at the right positions. |
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| | 3. Run 'Cytosoft 5.3' | | 3. Run 'Cytosoft 5.3' |
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| | 5. After cleaning, click Guava Express Plus on the left column. | | 5. After cleaning, click Guava Express Plus on the left column. |
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| - | 6. Go to the Analysis and Click 'Open Data Set' to set up the condition as our previous experiments. | + | 6. Go to Analysis mode and click 'Open Data Set'. |
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| - | 7. Go to folder 'iGEM' and open 'setting' file. | + | 7. Go to the 'iGEM' folder and open the 'Setting' file. |
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| | 8. Go to Acquisition - and hold here. | | 8. Go to Acquisition - and hold here. |
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| | 9. Go to the desktop and run WorkEdit 5.3. | | 9. Go to the desktop and run WorkEdit 5.3. |
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| - | 10. Highlight target sample areas. | + | 10. Highlight the wells you are going to use, check "Acquire this sample". |
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| - | 11. Make the condition as Guava Express Plus and check the box of '3 sec' and setup the speed from high to medium. | + | 11. Label them as Guava Express Plus, check the "Mix for 3 seconds", set the speed from high to medium. Optionally, fill the sample ID and the dilution factor. |
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| - | 12. Save and go back to the Cytosoft 5.3. | + | 12. Save and go back to Cytosoft 5.3. |
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| | --------------------------------------------------------------------------------------------------------- | | --------------------------------------------------------------------------------------------------------- |
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| - | 13. Start the worklist on the 'Go to Acquisition'. | + | 13. Go to Acquisition, start the worklist. |
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| - | 14. Locate the 96 well on the machine.
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| - | 15. Name our file as 'Today's date_title'
| + | 14. Place the 96-well plate into the tray, make sure the A1 well is where it should be. |
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| - | 16. When you are asked to adjust setting, check one circle of the 96 well for the control (Sowmya will give seed control).
| + | 15. Name the file as 'Today's date_title' |
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| - | 17. Compare with worklist and make sure that the conditions are the same as we made.
| + | 16. When you are asked to adjust the settings, check a well that contains seed cells. |
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| - | 18. After completing the adjust setting, click 'Resume' button.
| + | 17. Compare with the worklist, check if the flow and the amount of cells detected are reasonable. |
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| - | 19. Wait around 10~15min till the end - data will be saved automatically.
| + | 18. Click "Next step" and then "Resume". |
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| - | 20. After finishing the experiment, take out the 96 well and change the bleach position for the cleaning mode.
| + | 19. Wait until all the wells are measured - data will be saved automatically. |
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| - | 21. Go to Main menu and click 'clean and shut down'.
| + | 20. Take the 96-well plate out and insert the tubes that are required for cleaning. |
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| - | 22. Now you done :)
| + | 21. Go to Main menu and click 'Clean and shut down'. |
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