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Step 1: Use the Script Editor index sorting example from the Daily Dongle Blog (or see Addendum below regarding the method in Version 9). Running the initial script to create 96 populations Heatmap analysis of index sorting file.įigure 2. I'd really love for someone to tell me there is a way easier way to do this in FlowJo.įigure 1. What follows below is A method (not THE method) I stumbled upon to go from an index sort file from FACSDiVa to Figure 1 below.
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The goal of index sorting, in this case, is to retain the original FITC intensity information for each well after the sort. The resulting plate will be a mix of FITC low and FITC high clones. And here's the fun part.Īnalyzing index sorting data in FlowJo. I have an application where a user is index sorting based on a range of FITC intensities. There is a decent interface for looking at your index sort plate information within FACSDiVa, but if you're use to doing all your analysis is FlowJo, you probably want to bring that data over at some point. You can export both (or all) of these as FCS files. You should have a Pre-sort FCS file of your entire population, and an Index sort "tube" for each plate you ran.
Flowjo 10 vs flowjo 9 how to#
So, let's assume you figure out how to successfully perform an index sort in FACSDiVa. The second resource is a document presented at GLIIFCA 2014 by Matt Cochran (University of Rochester), in which he outlines some of his tips and tricks for working with index sorting in FACSDiVa 8. I only came upon this after our BD service engineer sent me a copy of it.
Flowjo 10 vs flowjo 9 manual#
The first, oddly enough, is BD's very own Index Sorting Manual (<- fixed bad link), which comes as an addendum to the FACSDiVa software manual and may not even be installed on your computer or available for download from BD's website. There are two resources that are quite helpful in figuring this part out. You may think index sorting is no more than checking a box in FACSDiVa, but there are enough one-off situations that arise that it really warrants a separate FAQ. Of course, I'm sure there are more elegant ways of doing this in other programs or even in FlowJo, but I needed this info yesterday, so I'm documenting it here for future reference.įiguring out index sorting in FACSDiVa. I've done the leg work already, so read on to get the info. They are out there, it's just a pain to jump around to various sites trying to compile all the information together. After scouring the web for resources on both index sorting and analyzing index sorting data outside of FACSDiVa, I decided to compile all the resources in one place. Getting used to a brand new set of DiVa issues and quirks has been difficult, but we soldiered on nonetheless.
Flowjo 10 vs flowjo 9 windows 7#
We recently upgraded our FACSAria to FACSDiVa 8 running on Windows 7 primarily for the ability to do index sorting.