Multiplex systems image the same sample beyond the traditional 3-5 labels. This has the advantage of profiling precious tissue with many markers, which can then be analysed at the single cell level, but also in relation to surrounding cells and tissue. PhenoCycler-Fusion 2.0 The Akoya PhenoCycler-Fusion 2.0 is a multiplexing slide scanner that can scan up to two slides at a time. Multiplexing works by hybridising a sample with a large number of barcoded antibodies (more than the traditional four or five) at once. Barcodes are hybridised three at a time and the whole tissue is scanned. Using a microfluidic robot, the PhenoCcycler cycles through steps of washing, new set of barcode hybridisation, and scanning. As a result, you get the same sample labelled with many antibodies (can go up to 100 on a single sample, with limitations). Results can be analysed with VisiopPharm, for which we hold a licence, or with the open-source software QuPath. Contact us before planning an experiment with the PhenoCcycler as the antibody sets and the dye-associated barcodes require appropriate pairing and optimising. We have successfully stained samples that were RNA-profiled on 10x Genomic’s Xenium, run by our colleagues of the Single Cell & Spatial Biology facility. Special applications and features: Multiple sets of antibodies (up to 100) on the same tissue sample One or two slide whole sample scanning Spectral unmixing of noisy samples (requires single dye controls as well as no dye control, unless using Opal dyes) Possibility to combine with in situ RNA profiling Location: IRR Imaging Hub, contact IRR.Imaging@ed.ac.uk for access. This article was published on 2024-08-15
