Wednesday, July 22, 2020

Plate reader & flow cytometry tutorials

Over the last two weeks, I've given two measurement tutorials in the iGEM Summer Webinar Series, one focused on plate readers and the other focused on flow cytometry. Both are posted in a repository on GitHub, along with example data and code to help people get started with effective calibration and interpretation of these instruments.
  • The first tutorial, "Quantifying fluorescence and cell count with plate readers," starts with a general introduction to fluorescence and OD, including a comparison of plate readers and other types of instruments, factors affecting fluorescence, and how to pick colors based on excitation and emission spectra. The second block focuses on calibration of measurements for fluorescence and OD, and on debugging such measurements. Finally, the session ends with a discussion of how to interpret and debug calibrated plate reader data.
  • The second tutorial, "Quantifying fluorescence and cell phenotypes with flow cytometry," starts with an introduction to flow cytometry, including how these instruments operate and the types of data that they produce. The second block focuses on calibration of measurements for fluorescence and cell size, and on debugging such measurements. Finally, the session ends with a discussion of how to interpret and debug calibrated flow cytometry data.
Under the hood of a flow cytometer, showing its optical path.
I hope that you will find these useful and redistribute them to others who may find the same!