Cytometry and Antibody Technology

New “Online” Tools from Coulter… Almost Useful!

by | Oct 13, 2009 | Archives | 0 comments

This archived post was originally written by Ryan Duggan when he was the Technical Director. Ryan has since moved to a position outside of the university. 

If you hop on over to the flow cytometry centric Beckman Coulter portal, aptly named coulterflow.com, you’ll see a link to some online tools. Tip #1: These aren’t actually online tools. They are downloadable, standalone programs which pull information from the coulter web site as needed. Tip #2: They’re pretty crude at this point. I can definitely see the utility they may bring in the future, but as of today, they need a lot of work. So, what are they? One is a fluorescence spectrum viewer, called Spectrios (a little corny, but hey, they’re rebranding themselves), and the other is an Experimental Design Tool called Experimental Design Tool, but I’ve renamed it to ExDT. Spectrios does pretty much what you’d expect it to, but again, it doesn’t seem ready for prime-time yet. The good thing about Spectrios is that it’s a standalone program, so you don’t need an active internet connection to use it.

That being said, it’s not nearly as good or as informative as the Invitrogen Spectra Viewer or the BD Spectrum Viewer (both online tools, though). Spectrios gives you the basics; Absorption curves, emission curves, laser lines, and filters. It does not calculate theoretical compensation nor allow you to add custom laser lines/filters. It is also lacking many fluorochromes from the list, but surely that will evolve over time. Spectrios and ExDT are so new and unpolished that they even forgot to take off the ***Concept – Beckman Coulter Confidential*** text in the header bar, and as is probably evident by now, these tools are still in Beta (v.0.9.0.7657).

Now here’s where things get exciting. ExDT is a tool that will (once it’s finished) greatly change the way you design your flow experiments. Built into the design tool are stock instrument configurations (laser lines, filters, # of detectors, etc…) and the entire Beckman Coulter catalog of Antibody/Fluor combinations. So, there are basically 4 steps to walk through (Screen shots below). Step 1 is a basic description of your experiment… Name, date, cell type, descriptions, etc. Step 2 allows you to select from the Beckman Coulter online catalog which markers you want to use. Now here’s the fun part. As you go through and select which markers you want to use, the panel at the bottom shows you all the available Fluors for the markers, and tells you how many possible combinations can be put together to make your panel. Also, if you wanted to force one of your markers (say CD4) to definitely be PC7, for example, that will automatically narrow your possible combinations greatly. So, if you know that the CD4 PC7 is a really good antibody, and you definitely will use, just Pin the antibody to that fluor, and now you may go from 30 possible combinations to 6. Step 3 then, simply allows you to flip through the possible combination and choose your panel. As you flip through the combinations, you get a “Spectrios-like” window showing you the emission curves so you can determine which panel you like based on available filters, or reducing compensation. Does it rank the combinations? Not sure if it does now, but that’d be cool. Does it allow you to upload your own catalog of antibodies? Not sure, but can you say awesome! Does it allow you to input your own instrument configuration with laser lines and filters, and then pick antibody/fluor combos to maximize sensitivity and minimize overlap? No, but if it did, I may faint with excitement. Finally, Step 4 allow you to set up your run list; Single stain controls, FMO controls, Sample tubes, etc… Oh yeah, you can conveniently click the “Add to cart” button just in case you don’t have those antibodies on hand and you need to purchase them from “you-know-who.” I call that Marketing Genius! Once you’ve made your run list, you can print it out and send it to your local Flow Cytometry Guru to give his/her blessing, and away you go. This tool has so much potential, I really wish it was working to its fullest right now. I have all these ideas swirling around my head on how I could use this yesterday. I guess we just play the waiting game now.

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