( included 384-well plates). The calculat...)measurement of green fluorescent proteins and a red fluorescent protein in live cells in the Analyst GT multimode reader

included 384-well plates). The calculation for the LLD was: 3SD_Blank /slope, where SD_Blank is the standard deviation of blank wells (media without cells) and the slope is the slope of the curve at the bottom end. (For the 96-well and 384-well plates respectively, the 10,000 cells/well and 2500 cells/well data points were used.) The RFU signal and standard deviation of wells containing non-transfected cells were similar to those containing media only.

The ZsGreen cell line had similar limits of detection in both experiments, whereas the other two differed as much as threefold between the experiments. We attribute these differences to the fact that their rates of growth, and therefore the number of passages, differed between cell lines and between the experiments. (With each successive passage, the cells become dimmer.) For example, in the first experiment, the DsRed cell line had 2-4 more passages than the other cell lines because it was growing more quickly. For better reproducibility, it would be better to design the experiments to keep the same passage number.

CONCLUSION
The Analyst GT multimode reader offers superior performance for measuring fluorescent proteins in intact adherent cells. Bottom reading gives better results, though top-read results are also quite good. The Analyst GT reader achieves its excellent performance with SmartOptics, a unique system that allows you to select and configure all components of the optical system to give you the best results for your assay. SmartOptics includes a speedy xenon flash lamp, as well as a powerful xenon continuous lamp so the correct light source can be used based upon assay mode. Also utilized in Analyst are high-quality fluorescence filters and fiber optics. Analyst has precise 3-axis positioning so that light is focused into individual wells and efficiently collected for maximum signal-to-background and sign
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