. . . the data always gets prettier at closing time . . . But seriously, what we're talking about here is holding ourselves to a higher standard - - and what's wrong with that? We've had a month's worth of discussion over good/bad publications, but nobody's looking at the cause . . . simply that there are confused operators/users in the field - - and many don't even know they're confused. Blame is not what we're about, but rather, what can we do to eliminate as much of the confusion as possible? Certainly training is key, courses are a plus, but ultimately we need a commitment from those of us who operate instruments to understand our technology to the maximum extent of our ability. Back in the day, when everyone had an "open" bench, understanding came from hands-on manipulation. With the development of turnkey analyzers, we lost that basic connection to the bench's parts, and with it the understanding of how those parts are important to the process. I've complained about the use of the arbitrary "FL" designations in the past as well, not because the questions are trivial, but rather that it represents a basic lack committment to understanding. When someone asks a question about why propidium iodide, when read in "FL2," bleeds into "FL3," it's obvious they don't really know what FL2 and FL3 are. If that's true, how can they be expected to use the flow cytometer correctly? Ultimately, I feel that part of the good/bad data problem lays somewhat with our failure to be harsh when it's warranted. In general, one rises to the level of the competition . . . and greater demand calls for greater effort. MAK. -- Mark A. KuKuruga, Managing Director University of Michigan Flow Core 7416 CCGC 0946 (734) 647-3216, fax (734) 936-7376 kukuru@umich.edu >>> Andrew Beernink <ABeernink@novasite.com> 12/14/01 04:15PM >>> True. But can anybody explain why I get better results after consuming beer Brand X or martini Brand Y? -----Original Message----- From: Mark Kukuruga [mailto:kukuru@med.umich.edu] Sent: Thursday, December 13, 2001 8:00 AM To: cyto-inbox Subject: Re: FACS question Paul . . . thanks for this response. I've tried this, and it works well. I've had some trouble finding a good source for reagent B though. MAK. -- Mark A. KuKuruga, Managing Director University of Michigan Flow Core 7416 CCGC 0946 (734) 647-3216, fax (734) 936-7376 kukuru@umich.edu >>> "J.Paul Robinson" <jpr@flowcyt.cyto.purdue.edu> 12/11/01 09:20PM >>> Colleagues: I am going to answer this question in a manner that I hope you all understand (or not)......(think of the bad data issue) You can solve the problem by adding reagent A to reagent B, plotting FL3 Vs FL2. If you add reagent C, then a single histogram of FL5 should do the trick. I think you could also try probe A and probe B, both of which should have the right spectra...Oh, turn the power of laser 1 up a bit, and you should get some valuable data..... sorry, I couldn't resist it... please stop using terms which are totally undefined.... Paul Robinson On 10 Dec 2001, at 11:58, Laura H odges wrote: I'd like to post a question on your website: Can anyone suggest another P-gp accumulation probe other than rhodamine 123 which has too broad of an emission spectrum for my use. I am trying to find a P-gp probe that can be used with other fluorescent markers in channels FL2, FL3, and FL4 in a multicolor assay. __________________________________________________ Do You Yahoo!? Send your FREE holiday greetings online! http://greetings.yahoo.com J.Paul Robinson, PhD PH:(765)4940757 Professor of Immunopharmacology Professor of Biomedical Engineering Purdue University FAX:(765)4940517 EMAIL:jpr@flowcyt.cyto.purdue.edu WEB: http://www.cyto.purdue.edu
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