Flow Cytometry Home

Plant Sample Prep for Nuclei Analysis

We don't have too many Botanists coming through the flow lab doors, but occasionally they'll bring in some electric green looking slurry of stuff and want to analyze DNA content or Mitochondrial Membrane Potential. It's really no different setting these samples up on the instrument than any other animal/human sample. But, when they start to ask me questions on how to optimize their sample prep, I'm of little help. From what I understand, the process is much like dissociating any tissue. You use some mechanical (slicing and dicing with a razor) and enzymatic processes to try and isolate the cells from the leaves. Beyond that basic understanding of what's going on, I have little to offer, which is why I appreciate this troubleshooting guide so much. A fella by the name of Paul Kron, from the Department of Integrative Biology at the University of Guelph (Guelph, Ontario, Canada) sent me his top 10 troubleshooting tips for achieving good quality histograms when analyzing DNA content (click on title for more...)

Submitted: 13 November 2009, 9:12 am

Do you Kaluza?

Seems like there's quite a bit of news coming out of Miami these past few months, and that is a good thing. Beckman Coulter (coulterflow.com) has released it's offline analysis tool, Kaluza, whose major feature seems to be speed. I've been playing around with Kaluza for about a month now and so I can share a few thoughts. You're probably well aware of the fact that we at the University of Chicago are a FlowJo shop. Nearly all of our users analyze with FlowJo (others use acquisition programs like CellQuest or FACSDiVa to do analysis). We've looked into other packages in the past (namely FCS Express and VenturiOne) but none were as exhaustive in capabilities as FlowJo, so we felt we were getting the most bang for buck in that case. What this basically me (click on title for more...)

Submitted: 9 November 2009, 6:53 am

New "Online" Tools from Coulter... Almost Useful!

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.
Imaging Indifference
I can't really get excited about imaging data. I don't know why, they're always really pretty pictures, but today I think I figured it out. Here's how a talk including imaging data typically goes. Slide 1: here is cell line x stained with Nuclear Dye A, and here is the same cell line stained with Membrane Dye B. Slide 2: now let's look at those two images super-imposed. (Ooos and Ahhhs radiate from the crowd). What do you do with that? How does this translate into some sort of metric than can be quantified and then objectively compared to a different situation or cell line. Flow Cytometry, on the other hand, is very quantitative, easily comparable, and believe it or not, objective. I understand the value of microscopy images, but I always feel like it should be validated and quantitated using some analogous technology like Flow Cytometry. Sorry Imaging folks... You may be asking yourself, well, if you're indifferent to imaging, why are you getting an ImageStream? Aha! Her (click on title for more...)

Submitted: 3 October 2009, 3:18 pm

Pre-GLIIFCA Core Manager Meeting

Well, another successful Core Manager Meeting just wrapped up here in PIttsburgh, PA. Topics presented ranged from Core Facility management, and cost recovery, to training issues, SOPs, and writing effective SIGs. Many of the talks recapped events from the NCRR/NIH Workshop in mid July. The overall meeting was entitled "Efficiency and Quality in the Management of Core Facilities: Best Practices for Running Your Core as a Business." I can honestly say, this Core Managers' Workshop provided some of the most useful information I've seen in years. It was great to hear many other core facilities around the country (and world...since we did have one attendee from Denmark) run into the same issues we do at the University of Chicago. We got some great ideas that we'll reevaluate once we get back home and see if we can implement a few of those things
Flow Cell

The Flow Cytometry Facility at the University of Chicago provides instrumentation, education, and expertise for the University's Faculty. The mission of the facility is three-fold:

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  • Provide instrument and applcation training and education

We encourage you to explore our services outlined in this web site.

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