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Counting Non-Aqueous Samples in the TopCount Microplate Scintillation and Luminescence Counter

ermined using 3H and 14C hexadecane for both the 96- and 24-well formats. To test the effects of sample load, varying volumes of samples in an organic solvent from an enzyme inhibition assay were counted in 96-well solvent resistant PicoPlates, and the results were compared to those obtained by conventional LSC. A TSH immunoradiometric assay was counted, and the results were compared to those obtained by gamma counting.

Experimental Methods

Counting efficiency was tested by adding 3H hexadecane to three wells of a 96-well PicoPlate containing 250 μL of MicroScint-O. For the 24-well PicoPlate, 3H was added to three wells containing 1.0 mL of cocktail. The same number of wells containing only cocktail were used for background determination. Similar experiments were carried out for 14C. The plates were sealed with Packards TopSealTM-P, a solvent resistant cover film, using the Packard MicroMate 496 Microplate Heat Sealer, and then mixed on a microplate shaker for 15 minutes. The samples were counted for five minutes each, under conditions optimized for liquid counting, on a TopCount with the VariPlateTM option which allows counting of either 96- or 24-well plates. To test the effects of sample load, 3H labeled samples in heptane solvent from an enzyme inhibition assay were counted using four different sample volumes. Aliquots of 25, 50, 100, and 150 μL in quadruplicate were pipetted into a solvent resistant 96-well Pico- Plate with a multiple-tip pipetter. Total volumes were brought to 300 μL with MicroScint-O. The plates were sealed and counted as described above. An additional 50 μL aliquot was counted for five minutes in a Packard Tri-Carb 2250CA liquid scintillation analyzer, using 5 mL Opti-Fluor organic cocktail in 7 mL glass vials, and the results were correlated with those from TopCount
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