Fluorescence imaging plate reader screening

The method may also be used the other way round with GOx as the enzyme label, glucose as substrate, and [Eu(Tc)] as the fluorescent indicator. In that type of assay, the presence of enzyme-labeled antibodies is indicated by an increase in the observed fluorescence intensity. It should be noted that commercially available fluorescence microplate readers, preferable equipped with time resolution, are also suited for the screening of all the microwell plate-based assays presented in this chapter. Nevertheless, the imaging process is much faster, accomplished in the order of one second, and enables ratiometric measurements. 

Zeiss have developed a high-throughput reader system (the Plate Vision multimode reader) for microwell plates that is capable of fast time-resolved fluorescence (Fast-TRF) imaging with nanosecond resolution [ 189] (Fig. 28), which has found applications in pharmaceutical screening. This microplate imager is suitable for time-resolved RET assays. Kinase assays, for instance, can be performed with antibodies labeled with a Ru(batho)2bipy complex or... 

Fig. 9.5 Simultaneous identification of binders of two antigens on a single filter, (a) scheme of the screen with two antigens each labeled with a different fluorophore. (b). Similar numbers of colonies from two independent selections (done on each antigen separately) were plated on the Supor master filter (the left half in (b)). The antibodies that diffused down into the cellulose acetate filter were developed with the fluorescently labeled antigens mixed together at 10(lg/ml each. The filters were washed and analyzed using a Typhoon 8600 reader (Molecular Dynamics). The right-hand half in (b) is a mirror image of the left filter with bright spots were green and dimmer spots marked by arrows were red...

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