Fl1

Figure 3.3. Each vertical strip in the top panel is from an individual well of a 96-well plate loaded with fluorescent beads. Two rows, 24 wells, of the plate are shown. The bottom panel represents a one-dimensional histogram for a single well. Data was collected on a Cytomation MoFlo FACS equipped with a Moskito autosampler.

mation MoFlo FCM equipped with a Mosikito autosampler, which is capable of sampling a 96-well plate in min. Each vertical stripe in the upper panel of Fig. 3.3 shows the side scatter for a single well and the lower panel is the FL1 histogram for a single sample. High-throughput compound screening by flow cytometry provides both a novel set of issues regarding assay cost, compound usage, dista handling, and data interpretation and novel cell pharmacology assays. All of these issues are under consideration in the development of the next generation of HTflow instrumentation.

Introduction of compounds to cells in flew provides another dimension to standard flow cytometry. The HTPS system described earlier has been adapted for flow cytometry aind increases the information content and expands the repertoire of assays, including high content toxicology data, available to the phar-macologylmedicinal chemistry drug development team. Early physiological responses such as changes in pH, intracellular calcium, and membrane potential can be monitored, and this can be coupled to cell-type specific markers, allowing assays to be performed on mixed cell populations. The HT-flow system has been used to measure cellular responses to test compounds at rates of 3-4 compounds per minute as well as determine the relationship between receptor occupancy and cell response (35).

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