Difference between revisions of "Tracks/Measurement/Interlab study/Flow Cytometry Overview"
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<h3><a href="http://parts.igem.org/Help:InterLab_Measurement_Kit">Go to InterLab Measurement Kit Information Page</a></h3> | <h3><a href="http://parts.igem.org/Help:InterLab_Measurement_Kit">Go to InterLab Measurement Kit Information Page</a></h3> | ||
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In order to carry out this measurement protocol, you will need: | In order to carry out this measurement protocol, you will need: | ||
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| − | <li>A flow cytometer with a channel configured for measurement of fluorescein (FITC) or GFP. This channel will typically have a 488nm laser | + | <li>A flow cytometer with a channel configured for measurement of fluorescein (FITC) or GFP. This channel will typically have a 488nm laser and a 530/30 filter, but the details may vary.</li> |
<li>Fluorescent calibration beads that have been calibrated for Molecules of Equivalent FLuorescein (MEFL) or Molecules of Equivalent GFP (MEGFP). If available, we recommend using SpheroTech Rainbow Calibration Particles RCP-30-5A, as that is what the accompanying spreadsheet has been designed for. </li> | <li>Fluorescent calibration beads that have been calibrated for Molecules of Equivalent FLuorescein (MEFL) or Molecules of Equivalent GFP (MEGFP). If available, we recommend using SpheroTech Rainbow Calibration Particles RCP-30-5A, as that is what the accompanying spreadsheet has been designed for. </li> | ||
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<b>Important!</b> If you have used a flow cytometer to measure GFP for the InterLab, please fill out the following Google Form to the best of your ability: | <b>Important!</b> If you have used a flow cytometer to measure GFP for the InterLab, please fill out the following Google Form to the best of your ability: | ||
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Latest revision as of 16:45, 21 June 2016
Measurement with Flow Cytometry
This is a general guide for setting up your cells for flow cytometry readings in absolute units. These steps and questions are meant to provide a general protocol and we ask that you follow them to the best of your ability.
In order to carry out this measurement protocol, you will need:
- A flow cytometer with a channel configured for measurement of fluorescein (FITC) or GFP. This channel will typically have a 488nm laser and a 530/30 filter, but the details may vary.
- Fluorescent calibration beads that have been calibrated for Molecules of Equivalent FLuorescein (MEFL) or Molecules of Equivalent GFP (MEGFP). If available, we recommend using SpheroTech Rainbow Calibration Particles RCP-30-5A, as that is what the accompanying spreadsheet has been designed for.
All teams using a flow cytometer to measure GFP must finish the following items to fulfill the InterLab:
- Email your completed Excel file to measurement (AT) igem (DOT) org [note: the empty template file is provided below]
- Fill in the Flow Cytometer Form provided at the bottom of this page
- Create an InterLab study page on your team wiki (please use this format: https://2016.igem.org/Team:YourTeamName/InterLab)
Download the Excel Template file here: TeamName_iGEM2016_Flow_Cytometry_Worksheet.xls
You can see a filled-in example of the worksheet here
Important! If you have used a flow cytometer to measure GFP for the InterLab, please fill out the following Google Form to the best of your ability:
