While it is increasingly recognized that three-dimensional (3D) cell culture models recapitulate drug responses of human cancers with more fidelity than monolayer cultures, a lack of quantitative analysis methods limit their implementation for reliable and routine assessment of emerging therapies. Here, we introduce an approach based on computational analysis of fluorescence image data to provide high-content readouts of dose-dependent cytotoxicity, growth inhibition, treatment-induced architectural changes and size-dependent response in 3D tumour models. We demonstrate this approach in adherent 3D ovarian and pancreatic multiwell extracellular matrix tumour overlays subjected to a panel of clinically relevant cytotoxic modalities and appropriately designed controls for reliable quantification of fluorescence signal. This streamlined methodology reads out the high density of information embedded in 3D culture systems, while maintaining a level of speed and efficiency traditionally achieved with global colorimetric reporters in order to facilitate broader implementation of 3D tumour models in therapeutic screening.

The ability to custom-tailor surface plasmon resonances of metallic nanorods via their geometry has made nanorods a prominent platform for the design of novel nanomaterials with wide-ranging potential applications. Characterization of nanorod geometries within their native solution environments is essential for targeting nanorod properties to the specific application and for improved monitoring of nanorod synthesis.Here we use a custom-designed depolarized dynamic light scattering setup for measuring nanorod translational and rotational diffusion in situ. Using either straight cylinders or prolate ellipsoids as model geometries, we developed an approach to convert diffusion measurements directly into predictions of nanorod lengths and aspect ratios. Neither of these two geometrical models was able to properly reproduce real nanorod diffusion. Yet, these discrepancies mostly precluded reliable predictions of nanorod aspect ratios while derived nanorod lengths were within 10−20% of their values determined separately from transmission electron microscopy.