How To Analyze NHP Lineage Flow¶
Created by Ravishankar Madhu¶
Last updated: 2023-07-10
Description¶
This protocol outlines the basics of how to analyze your flow data, specifically for the RM007a - NHP Lineage Panel. It won't explain how to use the software but will give guidelines on the gating strategy.
Things to Note before starting¶
Good Workflow¶
Keeping your data organized is extremely important, especially when working with longitudinal studies after a transplant that can span the course of 2 years. Here are some tips to ensure you keep your data organized - Always export both the experiment and .FCS files from the Flow Cytometers downstairs to the Kiem Lab Server. Stefan has a very robust file library for the Flow Cytometers, use it for the export. - When it is time to analyze files, have a folder in your Lab Member Folder on the Server where you can store the Workspace Files and copies of all the .FCS files you are analyzing. - Process each experiment in a different Workspace File, and do different gating strategies in different workspace files. - Export the images you receive from the analysis as PowerPoint files and the tables you make as .csv files
Procedure¶
![[Pasted image 20230524144646.png]]
This is generally the accepted Lineage breakdown.
Next Steps¶
You will want to generate graphs in PRISM detailing the lineage composition (Gran, Mono, B Cell, NK Cell, and T Cell) for each timepoint you take.