Mass‐tag barcoding for multiplexed analysis of human synaptosomes and other anuclear events. Issue 9 (5th April 2021)

• Record Type: Journal Article
• Authors: Gajera, Chandresh R.; Fernandez, Rosemary; Montine, Kathleen S.; Fox, Edward J.; Mrdjen, Dunja; Postupna, Nadia O.; Keene, Christopher Dirk; Bendall, Sean C.; Montine, Thomas J.
• Is Part Of: Cytometry. Volume 99:Issue 9(2021)
• Page Start: 939
• Page End: 945
• Publication Date: 2021-04-05
• Subjects: barcoding -- CyTOF -- mass cytometry -- platelet -- red blood cell -- synaptosome -- SynTOF; Flow cytometry -- Periodicals; Imaging systems in biology -- Periodicals; Imaging systems in medicine -- Periodicals; Diagnostic imaging -- Periodicals; 571.605
• DOI: 10.1002/cyto.a.24340
• Languages: English
• ISSNs: 1552-4922

Abstract

Abstract: Mass‐tag cell barcoding has increased the throughput, multiplexing, and robustness of multiple cytometry approaches. Previously, we adapted mass cytometry for cells to analyze synaptosome preparations (mass synaptometry or SynTOF), extending mass cytometry to these smaller, anuclear particles. To improve throughput and individual event resolution, we report here the application of palladium‐based barcoding in human synaptosomes. Up to 20 individual samples, each with a unique combinatorial barcode, were pooled for labeling with an antibody cocktail. Our synaptosome protocol used six palladium‐based barcoding reagents, and in combination with sequential gating increased the identification of presynaptic events approximately fourfold. These same parameters also efficiently resolved two other anuclear particles: human red blood cells and platelets. The addition of palladium‐based mass‐tag barcoding to our approach improves mass cytometry of synaptic particles.