Open-Top Light Sheet Microscope
The Open-Top Light-Sheet (OTLS) microscope is tailored to the needs of pathology practice and will be available to use within NDORMS in Q2 2023. The system creates pseudo-H&Es with small-molecule fast-acting nuclear (TO-PRO3) and cytoplasmic (Eosin) fluorescent dyes to aid with prognosis.
The instrument will mainly be involved in the investigation of features of lethality and non-lethality in prostate cancer tissue at diagnosis, using light sheet three-dimensional (3D) microscopy with ProtecT diagnostic samples (Prostate Testing for Cancer and Treatment – Prostate Cancer UK).
The OTLS design achieves an impressive balance of performance and versatility in a single system for a growing number of cleared-tissue imaging techniques in which high-resolution characterization of localized subregions is desired.
The OTLS system, developed by Jonathan Liu et al., for the University of Oxford, will be installed with the non-orthogonal dual-objective (NODO), allowing for high resolution of large tissue samples/organs.
Using the NODO imaging path, the system can be tuned to have a lateral resolution of between 400 - 800 nm and an axial resolution of ~2.5 microns. The objectives are positioned below the specimen, which provides an unobstructed open top that enables 3D imaging over a large 12 × 7.5 × 1 cm (xyz) imaging volume (see image left).
The imaging volume is limited in z by the objectives and in xy by the mechanical limits of the motorized stage. The objectives are sealed into a monolithic imaging chamber through direct immersion or the use of a solid-immersion meniscus lens (SIMlens), which provides multi-immersion capabilities spanning the refractive index range of all current clearing protocols.
The system has been used with ECi, PEGASOS, SHIELD, Ce3D, iDISCO, ClearSee/DEEP-Clear and CUBIC clearing techniques. This allows for flexible clearing protocols depending on the type of tissue sample or animal model.
The system is expected to be installed and operational by Q2 2023. Please enquire about system access/use with the DPOC facility team.
Image: The hybrid microscope consists of 3 objectives located below the specimen. The illumination objective is combined with two imaging objectives for orthogonal dual-objective (ODO) and non-orthogonal dual-objective (NODO) imaging.