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Advanced ACE technology boosts signal amplification for improved single-cell protein detection


Researchers have been using flow cytometry since the 1950s to characterize immune cells in research studies and blood samples. Traditional flow cytometry uses fluorescent probes linked to antibodies to detect proteins on cells. However, the method is limited in detecting multiple proteins. In 2009, mass cytometry was introduced, allowing the quantification of 50 proteins in single cells, providing a more detailed analysis of cellular identities and states.

Now, a collaboration led by the Wyss Institute at Harvard University has developed a method to enhance the sensitivity of mass cytometry and image mass cytometry (IMC) using DNA nanotechnology. By applying a new signal amplification technology called “Amplification by Cyclic Extension” (ACE) to DNA barcodes linked to antibodies, researchers were able to detect more than 30 different proteins with high sensitivity.

The new method allowed for the quantification of rare proteins, investigation of tissue changes, and study of protein networks in immune cells. ACE also facilitated the visualization of cell types and tissue compartments in histological sections, revealing changes related to kidney disease pathology.

By increasing the number of metal isotopes linked to antibodies using ACE, researchers achieved a significant boost in sensitivity for mass cytometry. This advancement enables high sensitivity, high multiplexing, and high throughput analysis of single cells, providing a deeper understanding of biological processes.

The study also utilized ACE to analyze epithelial-mesenchymal transitions in cancer cells, T-cell signaling responses, and injury-induced T-cell paralysis. The team’s findings, published in Nature Biotechnology, highlight the potential of ACE-enhanced mass cytometry for studying single-cell variations and intricate protein networks in various biological contexts.

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