Reagents for Mass Cytometry

Abstract

Mass cytometry (cytometry by time-of-flight detection [CyTOF]) is a bioanalytical technique that enables the identification and quantification of diverse features of cellular systems with single-cell resolution. In suspension mass cytometry, cells are stained with stable heavy-atom isotope-tagged reagents, and then the cells are nebulized into an inductively coupled plasma time-of-flight mass spectrometry (ICP-TOF-MS) instrument. In imaging mass cytometry, a pulsed laser is used to ablate ca. 1 μm2 spots of a tissue section. The plume is then transferred to the CyTOF, generating an image of biomarker expression. Similar measurements are possible with multiplexed ion bean imaging (MIBI). The unit mass resolution of the ICP-TOF-MS detector allows for multiparametric analysis of (in principle) up to 130 different parameters. Currently available reagents, however, allow simultaneous measurement of up to 50 biomarkers. As new reagents are developed, the scope of information that can be obtained by mass cytometry continues to increase, particularly due to the development of new small molecule reagents which enable monitoring of active biochemistry at the cellular level. This review summarizes the history and current state of mass cytometry reagent development and elaborates on areas where there is a need for new reagents. Additionally, this review provides guidelines on how new reagents should be tested and how the data should be presented to make them most meaningful to the mass cytometry user community.

Hybrid Macromolecular Constructs as a Platform for Spectral Nanoprobes for Advanced Cellular Barcoding in Flow Cytometry

“…[ 42 ] Cellular barcoding using antibodies labeled to distinct metals was successfully applied by Mei for mass cytometry measurements using anti‐CD45 antibodies. [ 7 ] In mass cytometry, the number of detection channels is (in principle) up to 130 but limited by the available metal tags [ 43 ] to 50, whereas conventional cytometry with 3‐laser excitation uses 8–12 channels (clinical) [ 44 ] to 15 channels (research) and full spectrum flow cytometry extends the number of resolvable fluorochromes further to 18 parameters. [ 45 ] Barcoding six or more samples with distinct antibody‐label conjugates means using up to six channels (systems of 5‐choose‐2 or 6‐choose‐3) [ 7,15 ] which is well tolerated in mass cytometry but impractical in conventional and full‐spectrum flow cytometry.…”

Selenium-based metabolic oligosaccharide engineering strategy for quantitative glycan detection

“…Methodological adaption from chemoenzymatic glycan labeling 68 or Se-click reaction 69 will integrate the enrichment and quantification of glycoconjugates, which doubtlessly provide room for methodological improvement. In addition, if the SeMOE analogs were synthesized as Se-isotopically enriched derivatives, these probes are readily distinguished by ICP-MS and may find broader applications in temporal studies 70 . These concepts are being pursued in our laboratory.…”

Design Parameters for a Mass Cytometry Detectable HaloTag Ligand

“…In future designs of MCP reagents, minimizing potential nonspecific binding should be considered paramount, as if this can be reduced, longer labeling times with higher polymer concentrations become feasible. Reducing nonspecific binding continues to be a significant challenge in the design of MCPs, and further research using novel antibiofouling strategies is warranted …”

“…Reducing nonspecific binding continues to be a significant challenge in the design of MCPs, and further research using novel antibiofouling strategies is warranted. 45 ■ EXPERIMENTAL PROCEDURES Materials and Methods. All procedures and synthesis not included below can be found in the Supporting Information along with supplemental figures, spectra, and the derivation of eq S12.…”