Single‐Cell Analysis: Toward High‐Dimensional Single‐Cell Analysis of Graphene Oxide Biological Impact: Tracking on Immune Cells by Single‐Cell Mass Cytometry (Small 21/2020)

Abstract

The cover, inspired by a famous album cover by Pink Floyd, represents the transformation of the “dark” undetected signal side of graphene into a visible signal, thanks to a specific functionalization with ZnS‐doped AgInS2 nanocrystals, allowing its tracking on the complex mix of human peripheral blood mononuclear cells at the single cell level, as presented in article number 2000123, by Davide Bedognetti, Alberto Bianco, Lucia Gemma Delogu, and co‐workers. The authors thank Fabio Del Regno for the cover design. Image credit: Davide Bedognetti, Lucia Gemma Delogu, and Fabio Del Regno.

Immune Profiling and Multiplexed Label‐Free Detection of 2D MXenes by Mass Cytometry and High‐Dimensional Imaging

“…Computational tSNE analysis was performed as previously reported. [28,29] The tSNE plots (Figure 2a), heat maps (Figure 2b-d) and bar graphs (Figure 2e,f) indicated that all 2D materials were naturally visible at the single-cell level and interacted with a vast number of immune cell subpopulations.…”

“…We chose the sub-cytotoxic concentration of 50 µg mL −1 and the 24 h time-point on the basis of the results of the viability assays (Figure S3, Supporting Information) and to ensure noticeable changes in immune cell functionality in terms of cytokine release and gene expression modulation, according to our previous experience with graphene-based materials. [28,29] Using a panel of 61 metal-tagged antibodies (Table S1, Supporting Information), we identified 15 distinct immune cell types according to the expression profiles of several cluster of differentiation (CD) markers present on the cell surfaces (gating strategy in Figure S4, Supporting Information). Computational tSNE analysis was performed as previously reported.…”

“…[28] Because carbon is not trackable with mass cytometry, its detection has been enabled through functionalization with AgInS 2 nanocrystals. [29] However, the material functionalization itself may affect the toxicological profile, [30,31] and the cell interaction and uptake capabilities; [32,33] it may additionally affect the intrinsic features of 2D materials that are exploitable for therapeutic/bioimaging purposes, [8] thus possibly impairing their biomedical effectiveness.…”

V4C3 MXene Immune Profiling and Modulation of T Cell‐Dendritic Cell Function and Interaction

“…Computational T-distributed Stochastic Neighbor Embedding (tSNE) analysis was then performed. [66,87] Figure 5A reports all the human primary immune subpopulations detected. Figure 5B shows cell viability analysis reported as t-SNE plots and heatmaps for all the selected subpopulations.…”

“…Gating Strategy and Data Analysis and Visualization: The CyTOF data analysis was carried out accordingly to the methods described by Orecchioni M et al [66,87] and Bendall et al, [103] by using a gating strategy to exclude doublets, cell debris, and dead cells using Cell-ID Intercalator-Ir and LD. Specific PBMC subsets and subpopulations were assessed as reported in Figure S5 (Supporting Information).…”