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Counting Cytostar-T scintillating microplates on the MicroBeta JET

200 μM kynurenic acid. The 0.1 μM domoate/ 200 μM kynurenic acid was added to the wells as one solution and the agonist induced response measured by counting Cytostar-T microplates for 20 secs/well over 10 cycles.

RESULTS

Figures 1 and 2 demonstrate the advantage of using the MicroBeta JET to add the agonist to the wells of Cytostar-T scintillating microplates. Using the MicroBeta JET, the induced response can be followed immediately after addition of the agonist. Adding the agonist manually results in a lag time where the initial response to the agonist cannot be measured, giving a significant difference in counts obtained for domoate in the first counting cycle. The lag time can be overcome using the MicroBeta JET to perform additions of the agonist to the wells of the Cytostar-T scintillating microplates.

The immediate effect of adding the antagonist kynurenic acid is shown in Figure 3. Suppression of the agonist induced response was immediately seen on addition of the domoate/kynurenic acid solution to the wells of Cytostar-T scintillating microplates. This was shown as a decrease in signal which was dependent on the concentrations of agonist and antagonist used in the assay.

Manual addition of kynurenic acid would give a similar delay in the initial response, as shown in Figure 2, resulting in a significant proportion of the antagonist effect of kynurenic acid being lost in that time.

CONCLUSION

Addition of reagents to the wells of the Cytostar-T microplate using the MicroBeta JET allows the response generated by addition of both agonist and antagonist to be measured immediately. Previously, on addition of these reagents manually, it was not feasible to obtain several measurements within a 5 minute time period, which resulted in a lag time where it was not p
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