Sperm sorting etc

Geoffrey Osborne (Geoff.Osborne@anu.edu.au)
Mon, 20 Jun 94 11:50:12 EST

Hi Frank,
Thanks for the loan of the PMT. I've just put it in the post to you.
It was unused as, as it turned out we made a new detector for forward
fluorescence detector housing. On this same subject I've been trying to
develop a protocol to enable us to analyse and sort sperm just using a
single UV laser, without the necessity for a forward fluorescence detector.
My approach to this has been trying to utilize the pulse processing
capablities of the 'Star plus and thus get information about the orientation
of the sperm. In the "standard" type of sperm setup, only one detector, call
it Fl1, is used for collecting UV fluorescence signal for orientation, while
the forward fluorescence detector which also collects UV signal is the
criteria that is essentially used as the basis for sorting.
My question to you is this. If you have an object (sperm head)the
shape of say a elongated flat dinner plate, passing through the laser beam
length ways, would the area, or width of the signal provide information
about the rotation of the object about its longitudinal axis? My feeling is
that provided the object is less than beam width, then no information can be
gained. Despite this, by plotting area versus width of the FL1 signal we can
achieve a signal which resembles a "normal" FL1 vs. FWD FL plot.
Got any bright ideas? I've about had it with the whole thing. One
approach I haven't yet tried is taking area and width on both signals and
seeing if there is difference, perhaps like a post acquisition ratio thing.
Anyway any feedback would be appreciated.
Probably should CC this to the flow mailing list.

Thanks
Geoff
=====================================================================
Geoffrey Osborne | ____ __ o Ahh!
Flow Cytometry (FACS LAB) | __ `\ <,_
John Curtin School of Medical Research, | __ (*)/ (*)
Australian National University, | ==============|
CANBERRA, AUSTRALIA. | |--|
Email: Geoff.Osborne@anu.edu.au | |--|...
======================================================================

Home Page Table of Contents Sponsors Web Sites
CD ROM Vol 2 was produced by 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:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu