CHICKEN SPERM AND BIS-BENZAMIDE
hilliard@zookeeper.zoo.uga.edu
Fri, 13 May 94 17:54:41 EST
I'm afraid this may get lost in the pending stats discussion (keep it
going!) but I do have a vexing technical question: does anyone have any
experience with detection of bis-benzamide stained sperm? There is a lab
here that's trying to measure the number of bisbenzamide labelled sperm
that are contained in sperm host glands after art. insemination (chickens
in this case). I can find the sperm exciting with UV and collecting
thru a 464BP, and they form a tight group on scatter (log SSC vs FALS),
but when we run oviducal cells mixed w/ sperm (in vitro) or oviducal
samples from inseminated hens we get almost nothing in that scatter region
or with the proper level of fluorescence. The number of sperm is
infinitesimal compared to other cells.
I guess this is essentially a rare event problem--does anyone have any
ideas? The embarassing part is that they can see these sperm clearly on a
fluorescence scope, but flow seemed to be a more objective way of
comparing all of the sample rather than one frame of a slide. Now they're
considering counts of confocal images, but would much prefer a faster flow
approach. Thanks in advance for any info you can send,
Steve Hilliard
Cell Analysis Facility, Univ. of GA : "Be good and you will
internet: hilliard@zookeeper.zoo.uga.edu : be lonesome"
voice: (706)542-9474 :
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