Tellurium‐based mass cytometry barcode for live and fixed cells

Abstract

Mass cytometry is a revolutionary technology that allows for the simultaneous quantification of >40 different biomarkers with cellular resolution. The biomarkers are detected using metal-labeled antibodies as well as small-molecule probes of cell size, viability, and biochemical status. Barcoding is an important component of sample preparation because it reduces processing time, eliminates sample-to-sample variation, discriminates cell doublets, reduces the amount of antibody needed, and conserves sample. We developed a thiol-reactive tellurium-based barcode, TeMal. TeMal is nontoxic at working concentrations, compatible with metal-labeled antibodies, and can readily be applied to live or fixed cells, making it advantageous and complementary compared to existing barcoding reagents. We have demonstrated the utility of TeMal by barcoding microscale samples in situ to facilitate analysis of cells from an automated cell culture system using mass cytometry.

Cell-type-specific signaling networks in heterocellular organoids

“…3c ). Using this knowledge, we developed a custom 20-plex ( 6 -choose- 3 , doublet-filtering 25 , 26 ) thiol-reactive barcoding strategy using monoisotopic tellurium maleimide (TeMal) ( 124 Te, 126 Te, 128 Te, 130 Te) 27 and Cisplatin ( 196 Pt, 198 Pt) 28 that can bypass ECM proteins and bind directly to fixed organoids in situ ( Fig. 3d , Supplementary Fig.…”

“…Fixed organoids were washed ×2 in PBS and stained in situ with 10 nM Cisplatin ( 196 Pt, 198 Pt) 28 (a kind gift from Dr. Olga Ornatsky, Fluidigm) and 1—3 μM TeMal ( 124 Te, 126 Te, 128 Te, 130 Te) 27 barcodes for 1 hr on a rocker (barcoding matrices in Supplementary Tables 3 and 4 ). (Note: organoids can also be barcoded overnight at 4 °C.)…”

Rare earth elements (REE) in biology and medicine

“…Besides Ln, mass cytometry also employs reagents based on other elements with a molecular weight of 75-209 Da e.g. palladium and platinum isotopes (Mei et al 2015;Schulz and Mei 2019), heavy metal nanoparticles (Schulz et al 2017;Pichaandi et al 2019), ruthenium and osmium (Catena et al 2016;Budzinski et al 2019), tellurium (Willis et al 2018), and bismuth (Han et al 2017). The labelled antibodies are used to stain single-cell suspensions in order to stably bind to their antigens on the outer surface of the cellular membranes, or, after appropriate fixation and permeabilization of the cells, in the cytoplasm and the nucleus.…”

An Iodinated DAPI‐Based Reagent for Mass Cytometry

“…To incorporate cellular phenotype reporters and cell‐type barcoding in parallel with antibody panels, small molecules bearing heavy elements were pursued for MC and IMC™ applications. This led to the development of numerous scaffolds, such as iridium and rhodium organometallic intercalators for cell enumeration, [29] bifunctional palladium and indium chelates for cell barcoding, [30,31] cisplatin for live/dead cell staining, [32] osmium and ruthenium tetroxide salts for cell barcoding and cell size discrimination, [33] bulky alkyl‐derivatives of chelators for staining bacteria [34] and eukaryotic organelles, [58] 5‐iodo‐2‐deoxyuridine (IdU) for cell cycle analysis, [35] and a collection of organotelluriums for a variety of applications including live cell barcoding in heterogenous suspensions [36] and organoids, [37] amplification of low abundance antigens in IMC™; [38] and monitoring biological states such as hypoxia, [39,40] senescence associated β‐galactosidase activity, [41] and protein synthesis turnover [42] . With the updated collection of MC and IMC™ reagents, 40–50 parameters can now be measured simultaneously from individual cells.…”