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:
- Download the Excel File below (TeamName_InterLab_2017_Flow_Measurements.xls) and fill it out with your data. Rename the file [TeamName] to reflect your team's name.
- Email your completed Excel files to measurement AT igem DOT org
- Fill in the two Flow Cytometer Forms (Form 1 and Form 2) provided at the bottom of this page
- Edit the InterLab study page on your team wiki (edit this page in your wikis: https://2017.igem.org/Team:[TeamName]/InterLab)
Flow Cytometer Forms
Download the Excel Template file here: TeamName_iGEM2017_Flow_Cytometry_Worksheet.xls
Download the Dilution Calculation Sheet here: 2017 Interlab Dilution Calculation Sheet.xlsx