CyTOF supports efficient detection of immune cell subsets from small samples

Abstract

Analysis of immune cell states is paramount to our understanding of the pathogenesis of a broad range of human diseases. Immunologists rely on fluorescence cytometry for cellular analysis, and while detection of 8 markers is now well established, the overlap of fluorescent signals limits efficiency. Mass cytometry or CyTOF (Cytometry by Time-Of-Flight) is a new technology for multiparameter single cell analysis that overcomes many limitations of fluorescence-based flow cytometry and can routinely detect as many as 40 markers per sample. This technology provides tremendous detail for cellular analysis of multiple cell populations simultaneously and is a powerful technique for translational investigations. Here we present reproducible detection of immune cell subsets starting with as few as 10,000 cells. Our study provides methods to employ CyTOF for small samples, which is especially relevant for investigation of limited patient biopsies in translational and clinical research.

Keywords: CyTOF; Leukocytes; Mass cytometry; Multidimensional single cell analysis; Translational immunology; Translational research.

Figure 1. Detection of cell lineages in different starting numbers of PBMCs

PBMCs were labeled with markers of immune cell lineages and detected by CyTOF. Representative data from 1 × 10⁶ PBMCs shown by SPADE tree outline of cell lineages. The node size represents the number of cells from 10⁶ PBMCs and shading intensity represents CD3 expression level (A). The means ± sem for absolute cell numbers (B) and percentages of each cell lineage in PBMCs (C) detected from indicated starting numbers of PBMCs are represented; n = 4 independent donors.

Figure 2. Multidimensional analysis of immune cells from skin

Cells harvested from healthy skin were labeled with markers for immune cell lineages and detected by CyTOF. Cell subset representation uses analysis platform SPADE (Qiu et al., 2011); n = 3 independent donors.