[RASMB] XLA NEW Jumping wavelength problem
John Philo
jphilo at mailway.com
Thu Dec 13 21:04:00 PST 2001
XLA NEW Jumping wavelength problemKate,
There are two potential causes for this problem: software and hardware.
In velocity runs, it is true that normally the delay calibration causes the
monochromator to move prior to the first scan, but the software is not
supposed to instruct the monochromator to move during subsequent scans at
the same wavelength. However, at least in some versions of the firmware in
the instrument that hasn't been true, and it keeps telling the instrument to
'seek' the desired wavelength prior to each scan, causing the wavelength to
fluctuate as a consequence of this hunting. I haven't heard of such problems
with recent firmware, though. FYI I was told earlier this year that Beckman
was going to change the firmware and have it use the measurement wavelength
for the delay setting, but I don't know whether this has been implemented
yet.
My instrument sometimes exhibits a phenomenon similar to yours, but it
appears to be a hardware issue. In my case, usually there is a single shift
in wavelength at some point during the run.
What I believe happens is that there is slow 'creep' of the position of the
post that moves the monochromator. On my instrument (a very old one) the
theory was that the o-ring that forms the vacuum seal on that post had
gotten sticky. We did replace that and that may have reduced the rate of
incidence of this problem, but it still recurs occasionally. Possibly
temperature fluctuations could cause some motion too? Anyway, I do see this
sometimes (not always showing as fluctuations in the wavelength recorded in
the header, but sometimes).
So you might have your service engineer at least change and lubricate that
o-ring, and perhaps check out the whole motor-pot assembly.
John Philo
-----Original Message-----
From: rasmb-admin at rasmb-email.bbri.org
[mailto:rasmb-admin at rasmb-email.bbri.org]On Behalf Of Katharina Spiegel
Sent: Thursday, December 13, 2001 11:27 AM
To: rasmb at server1.bbri.org
Subject: [RASMB] XLA NEW Jumping wavelength problem
Dear RASMB Colleagues,
I am encountering a phenomenon with our XLA which hopefully one of you has
seen already and can help me out with practical advice.
We started to see a change in wavelength during velocity scanning despite
setting the instrument to continuous scanning of one single wavelength only.
Admittedly, "starting to see" means actually that we "started to look", so I
don't know exactly for how long we have had this problem. Please view the
attached scan records. Our service engineer remains undisturbed, since he
considers these changes "within specs", but pressed harder for an
explanation/solution he inquired internally and came back with the following
explanation:
Since the delay calibration was not switched off the instrument performs
it prior to each scan thereby routinely moving the monochrometer to 500nm.
Switching back and forth between 500nm and our scan wavelength of 220nm is
then causing the variation. He suggested to speed up the rotor to the
desired speed and run the delay calibration once, switch it then off (check
NO DELAY CALIBRATION under options) and then "start the run" Obviously, this
approach needed to be altered to 1) fill rotor with mock filled cell and
counterbalance 2) spin it up to run speed and acquire one scan with delay
calibration 3) stop XLA and reload rotor with sample cells 4) program run
conditions with delay calibration switched off 5) start velocity run.
Short of time I did it this way for my most recent velocity run, and the
problem nevertheless remained. I take this as further evidence that there is
a instrument problem -one which my service engineer apparently has not yet
seen or at least is reluctant to acknowledge as a problem. Therefore, I am
looking for something to tip him off as to what he should be looking for.
I truly hope some of you have some ideas/suggestions what to do about
this.
Kind regards,
Kate
Cell 1:
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cell 3:
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00088.RA3:R 3 04.0 60000 0023243 9.1320E11 220 1
00089.RA3:R 3 04.0 60000 0023494 9.2310E11 220 1
00090.RA3:R 3 04.0 60000 0023749 9.3320E11 220 1
00091.RA3:R 3 04.0 60000 0024006 9.4340E11 220 1
00092.RA3:R 3 04.0 60000 0024263 9.5350E11 220 1
00093.RA3:R 3 04.0 60000 0024520 9.6360E11 220 1
--
*****************************************************************
Katharina Spiegel, Ph.D. Research Assistant Professor
Off: 847-467-6533 Manager, Keck Biophysics Facility
Lab: 847-467-6534 Northwestern University, Dept. BMBCB
Fax: 847-467-6489 2153 N. Sheridan Road
Email: k-spiegel at northwestern.edu Evanston, IL 60208 USA
http://www.biochem.northwestern.edu/Keck/keckmain.html
*****************************************************************
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