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Direct Counting of Millipore MultiScreen Filtration Plates.

A2M060E2), which contains a Durapore PVDF 0.45 mm membrane. A standard competition curve was prepared, following the instructions provided with the reagents. After incubation, the ligand-receptor complex was separated from free radioligand on the MultiScreen vacuum manifold (Millipore Cat. No. MAVM09601). The plate was then dried and snapped into the adapter (Packard # PPN 6005178). 25 mL of MicroScint-O, a lipophilic LS cocktail designed for TopCount, was dispensed into each well using a multichannel pipet. After covering with TopSeal-S protective sealing film, the plate was loaded and counted directly in TopCount.

A second plate was set up in parallel. The individual membranes on this plate were punched, using the MultiScreen Punch Kit (Millipore Cat. No. MAPK8960C), into 12 X 75 mm gamma tubes and counted on a Packard Instrument Company Cobra gamma counter. Figure 1 shows the correlation of counting data for the plates counted on the two instruments, and Figure 2 shows the resulting standard competition curves.

Total ligand tubes were also prepared and counted in the Cobra. After conversion of all counting results to DPM, a Scatchard analysis was performed (Figure 3). The Kd and Bmax determined with the two instruments are compared to reference values in Table 1.

These results show that cellular receptor binding assays can be performed in the MultiScreen filtration plate and counted directly in TopCount. Counting efficiency averaged 31.4%, with crosstalk not exceeding 0.25%. Final results are equivalent to those obtained by gamma counting.

Cell Proliferation Assay .
For this assay, we also chose the MultiScreen HV plate (Millipore Cat. No. SA2M060E2). The plate was sterilized and treated with 25% Rat Tail Collagen, Type 1 (Collaborative Biomedical Products, Bedford, Massachusetts, #40236) to impr
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