| | Applications | | Archive ! (02/2005) |
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New Applications Featuring TRF
Delfia and ratiometric Lance Assays have been tested with highest sensitivity as shown in Figure 1! |  Systems Integration
Assay Development
New Detection Mode
Optical Design
Applications
Technical Data |
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| Figure 1: Delfia sensitivity of the plate::vision reader in 384 well plates, detection limit < 0.1 pM Eu | |
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| New Applications Featuring FP
Dr. Thilo Enderle from Roche in Basel, Switzerland, used the plate::vision reader for the detection of the NEN [FP]2TM assay kit.
Figure 2 shows the result for a dilution series of the cAMP standard. The NEN [FP]2TM fluorescence polarization cAMP assay kit is based on the displacement of fluorescence-labeled cAMP from an antibody by unlabeled cAMP. As the unlabeled cAMP substitutes the fluorescence-labeled cAMP, the value of the polarization decreases with increasing cAMP concentration.
In these experiments, the plate::vision reader enabled Dr. Enderle to reach excellent z'-factors of up to 0.95 and a large polarization change mP of 110 mP. The evaluation of the curve resulted in a EC50 of 27 nM with the literature value being 23 nM. | | |
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| Figure 2: EC50 curve of Fluorescence Polarization cAMP [FP]2 kit (NEN), Costar 384 glass bottom, 50 µl measured with the plate::vision reader by Thilo Enderle (Roche Basel) | | |
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| Comparison of Black and Clear Plates
The 96-well quasi confocal illumination of plate::vision also provides a more economic advantage: Due to essentially “zero crosstalk” between adjacent wells - which stands for best data quality - clear microplates can be used instead of the typical black view plates which are considerably more expensive. This feature is confirmed by the measurement of the absolute crosstalk, the linearity and the sensitivity for both types of plates (see Figure 3). | | |
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| Figure 3: Dilution Series of Fluorescein | | |
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