Posters

The prevalence and clinical implications, of mechanism-based CYP450 inhibition has placed greater emphasis on the early detection of compounds with this potential. We have developed and validated a high throughput reversible CYP450 inhibition assay using human liver microsomes and industry recommended probe substrates.

With the increasing application of high-throughput assays for the determination of the in vitro ADME properties of compounds in lead identification and optimization, there is a growing need for efficient and cost-effective methods for interpreting theresulting data to enable well-informed selection to be made for compound progression.

Traditional methods for High Content Analysis (HCA) use technologies such as flow cytometry and microscope-based imaging systems. Laser-scanning microplate cytometry has many advantages over these, and is more amenable for use in High Content Screening (HCS).

The activation of GPCRs is known to lead to the dynamic translocation of multiple signaling molecules or molecular assemblies during its signaling cycle, and in many cases cytoskelatal reorganization. Such a movement and/or reorganization results in dynamic redistribution of cellular contents, equivalent to dynamic mass redistribution (DMR), which can be monitored online in living cells using Corning® Epic™ system – a label-free and non-invasive biosensor system.

In this study, we have used an Acumen Explorer equipped with a violet 405nm laser in conjunction with a selection of shorter-wavelength amine-reactive fluorophores (Invitrogen, Molecular Probes) to demonstrate a greater utility for blue fluorescent probes within high content assays.

Here we present a web-based bioinformatics solution for the management and analysis of all areas of cell-based screen experimentation: the Laboratory Information Management System of the Technology Development Studio (TDS-LIMS). TDS-LIMS is a software package consisting of the Library Checker, Library Browser, Scheduling Kit, Screen Browser and Screen Publisher.

Here we present a rapid method for determining ERK and JNK activation, demonstrated using time course, concentration-dependence data and the effect of the MEK1/2 inhibitor UO126 on ERK activation obtained using the Acumen Explorer™ laser scanning cytometer analysing 384 well plates.

For improved screening capability, we have developed a cell cycle analysis method using an Acumen Explorer fluorescence microplate cytometer, capable of reading an entire 384 well microplate in about 10 minutes. The method can perform such analyses on fixed cells in situ, markedly simplifying sample preparation.

Here, laser scanning microplate cytometry has been used to provide an automated high content readout of the effects of cytostatic agents on colony formation. This approach determines colony number through the application of a spherical volume algorithm. This permits the differentiation of cytostatic effects where the number of colonies and size remains constant and cytotoxic effects where the size and number may be reduced.