The technology, termed DNA sequence-topology assembly for multiplexed profiling (DNA STAMP), is a 3D barcoding platform for multiplexed profiling of subcellular protein expression and distribution. It comprises three functional steps – cellular targeting, 3D barcode generation and multiplexed readout of expression and distribution. For cellular targeting, antibodies conjugated with unique short DNA strands (Ab-short DNA) are used to label specific cellular proteins. Next, 3D barcodes are generated in situ from the bound antibodies, through nanostructure-assisted ligation of DNA tetrahedron probes with localisation identifiers. Once ligated, the 3D barcodes unfold and dissociate to liberate a pool of diverse, linear STAMP barcodes. Each STAMP barcode thus reflects the target marker’s identity, quantity as well as its subcellular distribution pattern.
The device integrates three major functional components:
i) a serpentine mixer for efficient cellular targeting with antibodies,
ii) an embedded, porous membrane for rare cell enrichment and 3D barcoding of targeted cells, and
iii) multiple chambers for STAMP barcode collection, amplification and analysis.
The microfluidic platform could be loaded onto a custom-designed thermal cycling system for amplification of STAMP barcodes. All fluidic flow was controlled through torque-activated valves to streamline the assay operation.
- Disease diagnostics
- Rare cell analysis
The global anatomic pathology market is estimated to grow at a compound annual growth rate of 6% from 2019 to US$24 billion in 2026. The growth is mainly attributed by: