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The left right adjust knob on the old autoclone adjusts the bias on the
droplet charge neutral. The voltage across the plates always remains
constant. For this pot to work, your 753 sort control module (lower left
fron with the counters and drop drive adjustments) must have been modified
by Coulter. There should be a connector with 3 wires (gren,orange,white)
that plugs into the lower right corner in the back of your autoclone drawer.
If this is working properly you should be able to swing the center stream
about 5 degrees from left to right.
Getting the cells through the first hole is easy. We use the same system to
sort into the bottom of conical 96 well plates. To check alignment we made a
96well plate pattern using powerpoint and printed it out on a transparancy.
Then glued this to a plate. You can then sort on to the transparancy and
easily check alignment with an epi microscope. We can easily see individual
sperm labelled with 3342 using this method.
I presented a poster several years ago showing how you can modify a
cicero/sutoclone system so that you can program any number of cells for any
position on the plate. Let me know if you're interested in more detail on this.
>
>Hi folks,
>
>A few weeks (months?!) ago I asked about sorting single sperm for PCR,
>and recieved quite a few encouraging responses. Th interested party has
>obtained the dye (we're using Ho348) and tested their PCR after staining,
>and they're quite satisfied and ready to sort.
>
>Now the ball is in my court....
>We have a 753 with Cicero. I dug out our auto-clone unit, got the 753
>sorting beautifully with the normal sort drawer, and then installed
>the auto-clone. Now I'm stuck! The next step is calibration and
>testing, to adjust the instrument so that the sorted test stream
>(actually the "unsorted" stream) goes through a hole in a plate in the
>sort drawer. The multiwell plate is underneath this hole, and any droplet
>that is going to land in the well must go through this hole. Problem is, my
>stream won't go through the hole! The waste (what I would call the
>center stream and the left stream) is deflected to the left and lands in
>the waste catcher, but the stream I want (the right stream, which should
>be falling straight down) is too far to the right. The manual
>indicates that I should adjust the right-left and front-back knobs on the
>autoclone until the stream goes through the hole, but the
>right-left knob has no noticeable effect. Does anyone know what it
>does? It looks like a potentiometer, and I'm speculating that it
>modulates some bias in the plate charges to shift all streams
>right or left. I thought about yanking it and substituting one w/ a
>larger range of resistances, but there should be better way. I tried
>decreasing deflection gain, and that does move it a bit to the left, but
>not enough, and I'm limited by the fact that at too low a setting the
>waste streams start splattering off the edge of the waste catcher.
>
>Does anyone have any ideas? I won't be able to work w/ the instrument
>again for another week, but we really need to get this working.
>Suspecting that deflection in one direction may be higher than the
>other, I'm going to try switching the plate polarity and cable
>setups so that right is left. Does anyone have any experience w/ this
>beast, or know of any other tricks?
>
>-------------------------------------------------------------
>Steve G. Hilliard Join the fight--help
>Cell Analysis Facility stamp out smartass
>University of Georgia sig files!
>-------------------------------------------------------------
>
>
>
Gary Durack
University of Illinois Biotechnology Center
voice (217) 244-0559 fax (217) 244-0466
durack@uiuc.edu
http://www.life.uiuc.edu/biotech
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web