HTS-Compatible Assays to Assess Drug Effects in 3D
Most biochemical assays were developed for use on monolayer models and need modifications depending on the tissue type due to high secretion of extracellular matrix. Moreover, when using complex models at an early stage, the costs and read-out speed have to be taken into account.
Lytic assays, such as the CellTiter Glo (Promega), have shown compatibility with the microtissue format to assess tissue viability. However, to get the most out of a microtissue-based assay, additional nonlytic read-outs can be added to discriminate effects on distinct cell populations in the model. To this end, InSphero has developed a specific assay plate to culture microtissues at a defined position.
The plate is compatible with the Tecan Infinite M200 PRO, which captures the signal of a whole well without the need for multispot measurements. Together with the use of highly sensitive reporter systems, this system has become an efficient and automation-compatible tool for assessing drug effects in microtissue models.
The Infinite M200 PRO is a versatile multimode reader, based on a sophisticated double-monochromator optic; it offers high sensitivity for all fluorescence- and luminescence-based measurements. In a previous study, the optical properties of the system were shown to perfectly match the necessary requirements for sensitive detection of fluorescent signals originating from 3D microtissues.
In addition to the unique design of its fluorescence optics, the Infinite M200 PRO is equipped with a separate and independent luminescence optic, developed to detect luminescent signal with the highest possible sensitivity, which allows convenient analysis of luminescence-based reporter signals, like those of the NanoLuc luciferase.
The smart design of Tecan’s i-control™ software enables multiplex measurements like those used in this study. Tecan’s robotic workstation, the Freedom EVO®, is capable of handling 3D microtissues and the Infinite M200 PRO multimode reader can be integrated into the system to optimize the process for high-throughput analysis.