Geoff, If you can get 5 cells in each puddle on the lid every time, your single cells are probably going into the well every time too. To confirm this by inverted microscope, don't forget to wait for your cells to settle to the bottom; often they seem to sit at the edge of the meniscus for a while before taking the plunge. Round bottom wells are good for locating the cells (see Boezeman et al Cytometry 28(4):305-10 1997). If you prefer to see the single cells on a slide, the fast evaporation of the droplet will give you problems but see Alberti et al Cytometry 5(6):644-7 1984 and Kanz et al Cytometry 7(5):491-4 1986. Frank. Frank Battye, \ / < The Cytometry Laboratory Dora Kaminaris, \__/ <<<<< The Walter & Eliza Hall Institute Viki Lapatis,----------!!<<<<<<<< Victoria 3050, AUSTRALIA Jennie Parker, /!!\ <<<<< ph: 61_3_9345 2540 fax: 61_3_9347 0852 Catherine Tarlinton o !! \ < in:: "facs_copy@wehi.edu.au"
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