Soft X-ray tomography and structured illumination fluorescence microscopy as integrated high throughput pipelines for drug discovery, production and post-market surveillance
To meet safety standards, drugs must be safe, specific, and reproducible with unquestionable emphasis on public safety. Conventionally, at the exploratory and pre-clinical stages, methods employed for drug development are based on physicochemical parameters (for example proteomics, NMR, spectroscopy, calorimetry and ELISA). These traditional drug profiling approaches lack high-resolution 3D data on native-state biological carriers or their effects on host cells.
To better understand the action of pharmaceuticals in cellulo and develop a translational tool for vaccine development and cross-evaluation, we have developed a high-throughput correlative 3D imaging platform and pipeline for biomaterials and cells under near-physiological conditions at beamline B24, Diamond Light Source (Figure 1). Here, we provide a tool for industrial and academic partners to further validate drug suitability during research and development. This is achieved through a robust sample reparation protocol (including snap freezing to preserve native structures at physiologically relevant conformations) which was developed using model cell lines to ensure consistent imaging within cell populations.
Deliverable measurements include cell size and gross morphology, polarization of cytoskeleton and endosomal factors, change in size and distribution of organelles, nuclear membrane remodeling and exosome production. Effects can be captured and assessed using a fully commissioned correlative cryo-imaging platform and their potency as indicators of drug-induced side-effects can be scored.
We deliver a package of refined protocols ranging from sample preparation and handling to data collection and processing along with a preliminary catalogue of cellular features that we can assess to delineate the suitability of pharmaceuticals prior to further studies (Okolo et al., 2021. Journal of Microscopy. https://doi.org/10.1111/jmi.13054).