Wayne, I believe the problem lies in the cells size to nozzle size ratio. It should be at a minimum about 1:3 ratio with a 1:4 or 5 even better. When the embryo reaches the bottom of the sort stream, it causes the droplet to breakoff sooner then it should and therefore the saline droplet that has the cell in it is only partially charged. Another possibility is that the total charge on the droplet is lower because the saline volume of the droplet is lower due to the large embryo volume. If either of the above happens then the deflection of the partially charged droplet is suboptimal. The droplet never deflects far enough from the center stream to reach the collection tube. There is an old paper (1977) about particle size effect on the droplet breakoff written by Richard Stovel. Look up Journal of Histochemistry and Cytochemistry Vol. 25 No.7 pp813-820 Good luck. Glenn Glenn Paradis MIT Flow Cytometry Core Facility E18-463 50 Ames Street Cambridge, MA 02139 Voice: (617) 253-6454 Fax: (617) 253-3714 Email: gap@mit.edu
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