[FDS] Calibration strip

[Virgil Schirf]

Glen,

Thank you for the information. I re-degassed the fluorescein and 
refilled the strip, re-focused the signal, and the signal showed the 
plateau as you described. I was able to see the sample and it was being 
read right in the center of the chamber, so I believe the radial 
calibration is correct. But I still receive the 'calibration strip 
outside of size limits error.' Forcing the magnet angle after 
re-focusing doesn't correct the problem.  It seems to me that if you can 
view the calibration strip in the fluorescent focusing mode through the 
hardware/software, the magnet angle should be within limits, although 
I'm sure that's too simplistic.

Thanks again,

Virgil

Glen Ramsay wrote:

> Greetings Virgil:
>
> This happens occasionally.  The calibration strip is so tiny that it 
> is easy to have a bubble.  To complicate the situation the meniscus 
> changes shape depending on the speed, so the system may work at some 
> speeds but not others.  Ideally the strip is bubble free.  You must 
> degass the dye before loading the calibration center piece, otherwise 
> quenching will significantly weaken the signal.
>
> Another factor is the focusing.  If the system isn't focused the 
> signal is smeared and weakened. Refocus if you haven't already tried 
> this.  Click on the /Vertical Scan /button and the system will 
> automatically scan the focus.  If a well defined peak appears then 
> click on the /Save Focus /button.  Close the panel and use the 
> /Hardware /menu, /Force Magnet Angle... /option to see if the problem 
> is fixed.
>
> Adjusting the radial position can also help.  If the beam is touching 
> the bottom of the calibration strip, or the meniscus, then the signal 
> will be weakened or misshapen.  In the Focus Panel you will see a 
> Radial Position slide bar.  The default is 5.85 cm.  Making small 
> adjustments (0.1 mm) will allow you to see if there is a better 
> position.  The real time signal must show a clear plateau with no dips 
> or peaks, or rounded shoulders. 
>
> Occasionally the limits need to be adjusted, but since your system has 
> already worked I would think that they are OK.
>
> Glen
>
>
> At 05:21 PM 10/18/2006, you wrote:
>
>> Good afternoon,
>>
>> After several successful runs using the fluorescence optics, I am now
>> getting a calibration strip outside of size limits error. I've taken
>> apart the calibration cell, cleaned and refilled the strip several
>> times, making sure there are no air bubbles, etc; however, I continue
>> receiving the same error. Does anyone else have any suggestions for what
>> might cause the problem? I might add that I did 'see' the strip when in
>> fluorescent focusing, so I know that it's there.
>>
>> Thanks in advance,
>>
>> Virgil
>> -- 
>>
>> Virgil Schirf, Manager CAUMA & CLS
>> Department of Biochemistry, Rm 4.523U
>> The University of Texas Health Science Center
>> 7703 Floyd Curl Dr.
>> San Antonio TX 78229-3900
>> Voicemail: 210 567-6590
>> Fax: 210 567-6595
>> Email: schirfv at biochem.uthscsa.edu
>>
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>
>    Glen Ramsay, Ph.D.
>    Chief Scientist
>    Aviv Biomedical
>    750 Vassar Avenue
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>    www.avivbiomedical.com
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-- 

Virgil Schirf, Manager CAUMA & CLS
Department of Biochemistry, Rm 4.523U
The University of Texas Health Science Center
7703 Floyd Curl Dr.
San Antonio TX 78229-3900
Voicemail: 210 567-6590
Fax: 210 567-6595
Email: schirfv at biochem.uthscsa.edu