Best Practices Document for how to set up a cytometry run

[mehersam]

1.) Create the document 2.) Should be in the 'FlowCytometryDocumentation' directory 3.) Link it from gh-pages

Information to include:

Every experiment should include at least the following controls: SpheroTech URCP-38-2K beads or RCP-30-5A beads. Any lot will do, but please document the lot, since the calibration values differ significantly from batch to batch. Negative control One strong constituent positive control for each individual color for each experiment. One cotransfection of all colors in equivalent strong positive controls.

Here are the best practices for data collection: When gathering data, gather a set of bead data at least once per day, and also each time that you change the flow cytometer settings (e.g., channel voltage). Gather at least 30,000 events in your bead sample. There should be a clear set of "peaks" in the FITC channel. Document the laser/filter combination of your flow cytometer's channels. The preferred channels are: MEFL [“green” - FITC ERF]: Defined channel: 488nm laser, 530nm/40nm filter Alternate standard: 488nm laser, 530nm/30nm filter MEPTR [“red” - PE-Texas-Red ERF]: Defined channel: 488nm laser, 613nm/20nm flter Alternate standard: 561nm laser, 610nm/20nm filter MEBV421 [“blue” - Brilliant Violet ERF]: Defined channel: 405nm laser, 450nm/50nm filter Make sure you gather area data. Height or width can be included if desired, but don’t give the desired measurement. Make sure you are recording FSC-A and SSC-A, and that your hardware gating is fairly permissive.

[coverney]

If we just copy what Meher wrote, I can easily add this to our TASBE website.