CrestOptics SpA
Luca Clario, Francesco Bacchi
On Stand Workshop - Stand 8
Spatial -omics technologies enable a deeper understanding of cellular organizations and interactions within a tissue of interest. These assays can identify specific compartments or regions in a tissue with differential transcript or protein abundance, delineate their interactions, and complement other methods in defining cellular phenotypes. Imaging-based spatial transcriptomics mainly uses epifluorescence microscopy, which has shown remarkable results for the identification of multiple targets in situ. Nonetheless, the number of genes that can be reliably visualized is limited by the diffraction of light and by the concentration of fluorescence targets in the single FOV, also referred to as the optical crowding. Besides, spatial biology is gradually moving towards the transcriptomic profiling of thicker samples and up to whole organs and embryos where good optical sectioning capacity is required. Therefore, alternative methods to standard wide field imaging are rapidly sought for robust and reliable detection of each individual transcript. In this workshop, CrestOptics Spinning Disk Confocal and SIM (Structured Illumination Microscopy) technologies are proposed in a combined experimental setup to increase spatial resolution (minimum size of molecular units profiled), coverage (breadth of tissue covered), scale and throughput (number of samples and profiling speed), and multiplexing capacity (breadth of molecular entities profiled simultaneously). With Spinning Disk Confocal tile scan and Z-stacks of entire tissue sections are accomplished at high speed. SIM can help to untangle high-density areas and resolve numerous spots in close proximity at subcellular levels. Taken together, Spinning Disk Confocal and SIM have the capacity to improve spot detection and overall data quality in spatial transcriptomics with respect to standard wide field microscopy plus deconvolution approach.