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First of all, thanks to everybody who responded to my request for tips and
tricks on changing the dye. I changed the dye with the help of the field
engineer from Coherent, and it was much less of a headache than I thought
it would be. It took only a couple of hours, including 30 minutes to
dissolve the R6G in methanol. Just make sure you have all the materials and
tools you need, take your time and be careful. We didn't spill a single
drop!
I have one last question about some of the procedures I received. A couple
of people mentioned that I should measure the absorbance (at 514 nm) of the
dye when optimizing for dye laser output. I would appreciate any discussion
of what we are looking for when we do this. Also, what kind of blank to use
in the spectrophotometer: ethylene glycol, water or what? Is it necessary
to make any dilutions in order to get a reading on scale? Is the linear
part of Beer's Law important here? I'm guessing that I am looking for an
increase in absorbance of the dye up to a certain maximum and then a
decrease in absorbance to tell me that I am beginning to add too much dye.
Is this correct? I hate to keep re-inventing the wheel when there are so
many people out there who have already done a very good job of it for me.
:-)
Best regards,
Steve
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| | | | | | Steven Micko, Senior Flow Cytometry Specialist
| | | E | | | Cell Separator Dept, Emory University Hospital
| | | U | | | 1364 Clifton Rd, N.E., Atlanta, GA 30322
| | | H | | | voice 404-712-4357, FAX 404-712-4332
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\__||___||__/ steven_micko@email.eushc.org
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