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

. Dose curves for the enzyme inhibitor and a control sample value obtained with TopCount were compared with those from the Packard Tri-Carb 2250CA.

A sandwich immunoradiometric assay (IRMA) for TSH was performed in a solvent resistant PicoPlate or in microplate strips (Dynex Technologies) coated with anti-TSH MAb as the first antibody. An 125Ilabeled anti-TSH MAb was used to quantitate TSH bound to the solid support by the first antibody. The PicoPlate was counted in the TopCount with 250 μL MicroScint-O, and individual wells of the microplate strip were counted in a Packard Cobra gamma counter for comparison.

Results and Discussion

The average efficiencies and backgrounds obtained with TopCount with a small sample load are presented in Table 1. A small sample (10 μL) was used to provide the optimum cocktail performance. Counting efficiencies for 3H are comparable to those which can be obtained by conventional liquid scintillation counting.

The CPM results for equal volumes of 3H labeled samples in heptane counted with TopCount and a conventional LSC are compared in Figure 1. The data show that the TopCount correlates very well with the LSC (R2= 0.99). The slope of the line indicates that for this sample load TopCount CPM is approximately 40% of the CPM obtained by conventional LSC. This lower efficiency is a result of the much higher sample load; only 250 μL of cocktail was used in TopCount versus 5 mL in the LSC.

Table 2 shows a comparison of the % inhibition values for a control sample obtained from TopCount with the value obtained by LSC. Assays 1-4 were counted with TopCount using four different sample volumes, and assay five was counted with the Tri- Carb 2250CA. These control values are additional confirmation that TopCount provides consistent results regardless
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