25 Jun Functional proteome profiling of single cells using a microchip assay
Technological advancements in sequencing technologies, as well as high-throughput methods such as droplet microfluidics, have led to more widespread use of transcriptomics in single-cell research. However, not all single-cell omics are equally advanced. For instance, although proteins can be of utmost importance in disease diagnostics, single-cell proteomics is not yet well developed. For this week’s research highlight, we have selected a recent article published in Nature Communications aiming at improving the proteomics landscape. Using microfabrication methods and a PDMS-based microchip, the research team developed a single-cell cyclic multiplexed in situ tagging (CycMIST) platform and could successfully detect 182 proteins.
“The high protein content is achieved by the multi-round labeling and multi-cycle decoding process based on the CycMIST platform. Each single cell in a microchip microwell can be stained for 4 rounds by a cocktail of UV-cleavable DNA barcoded antibodies, and for each staining the DNA oligos can be released by UV irradiation and captured by a MIST array.“, the authors explained.
“Our microchip is amenable to integration with an instrument that can handle liquids and scan the entire array. Development of advanced data acquisition will be needed to standardize the process to be like flow cytometry, which has the hardware gating system and associated software to improve data quality and lower detection noise. A higher throughput and higher multiplexity will be achievable after further development to make the CycMIST as a tool complementary to single-cell sequencing.“, the authors concluded.
Figures are reproduced were reproduced from Yang, L., Ball, A., Liu, J. et al. Cyclic microchip assay for measurement of hundreds of functional proteins in single neurons. Nat Commun 13, 3548 (2022). https://doi.org/10.1038/s41467-022-31336-x under Creative Commons Attribution 4.0 International License.
Read the original article: Cyclic microchip assay for measurement of hundreds of functional proteins in single neurons