Single-cell fucosylation breakdown: Switching fucose to europium

Abstract

Summary Fucosylation and its fucosidic linkage-specific motifs are believed to be essential to understand their distinct roles in cellular behavior, but their quantitative information has not yet been fully disclosed due to the requirements of ultra-sensitivity and selectivity. Herein, we report an approach that converts fucose (Fuc) to stable europium (Eu) isotopic mass signal on hard ionization inductively coupled plasma mass spectrometry (ICP-MS). Metabolically assembled azido -fucose on the cell surface allows us to tag them with an alkyne -customized Eu-crafted bacteriophage MS2 capsid nanoparticle for Eu signal multiplication, resulting in an ever lowest detection limit of 4.2 z mol Fuc. Quantitative breakdown of the linkage-specific fucosylation motifs in situ preserved on single cancerous HepG2 and paracancerous HL7702 cells can thus be realized on a single-cell ICP-MS platform, specifying their roles during the cancering process. This approach was further applied to the discrimination of normal hepatocellular cells and highly, moderately, and poorly differentiated hepatoma cells collected from real hepatocellular carcinoma tissues.

Selenium-based metabolic oligosaccharide engineering strategy for quantitative glycan detection

“…11 ). Previous research based on Europium mass tag and bioorthogonal reaction gave the quantitative density of cell-surface newly-synthesized fucosylation at 5~100 × 10 -18 mol/cell 14 . Based on ICP-MS, we measured the Se signal at 100~2000 × 10 -18 mol/cell in various cancer cell lines, which was comparable in the orders of magnitude.…”

“…The Varki group has demonstrated this concept via radiolabeling approaches, but suffers from cost-ineffectiveness and radioactive safety risks 12 . Additionally, maneuvering MOE and installing the incorporated bioorthogonal functional groups with alternative signal output modalities, such as 1,2-diamino-4,5-methylenedioxybenzene (DMB) 13 , Europium (Eu) isotopic mass tag 14 and surface-enhanced Raman spectroscopy (SERS) probe, have allowed for the detection and quantification of monosaccharide derivates 15 . Nevertheless, the stoichiometric attachment of complementary probes to glycoconjugates is inevitably hampered by their accessibility, and the non-specific interactions between added probes and cellular components (e.g., cell surface) additionally increased the difficulty of quantitative readouts.…”

AI and the next medical revolution: deep learning’s uncharted healthcare promise

“…Liu et al [95] applied CNNs to model the spatial correlation structure of histone modifications and predict expression. Yang et al [96] developed a deep adversarial network for predicting methylation states from multimodal descriptors, including DNA sequence motifs and 3D chromatin structure.…”

ICP-MS-Based Methodology In Metallomics: Towards Single Particle Analysis, Single Cell Analysis, And Spatial Metallomics

“…73 Using a chemical labeling strategy, the fucose contents in single cells were determined via europium using scICP-MS, achieving a detection limit of 4.2 zmol fucose. 74 Single bacteria metabolically labeled by a lanthanide-encoding were counted and recognized with scICP-MS, providing a new way to identify bacteria and study variability at a single bacterium level. 75 Mass cytometry, a specific scICP-MS using a TOF analyzer, was originally designed to analyze single cells labeled with lanthanide isotopes.…”