Mass Cytometry Applications for
Cancer Research
Pub Note PN 13–02_150505
Contents
Cell cycle analysis of the human hematopoietic system by mass cytometry
4 Mapping phenotypic heterogeneity and progression of cancer cells by mass cytometry
5 High-resolution profiling of the baseline diversity in human NK cells
6 Bibliography
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Mass Cytometry Applications for Cancer Research
The CyTOF® Mass Cytometer uniquely isolates signal from over 30 probes into
single channels with minimal signal overlap, thereby enabling system-wide singlecell proteomic studies. Typical probes used are target-specific antibodies
conjugated to rare earth metals and targets studied include cell surface markers,
cytokines, signaling proteins and nucleic acids.
This document highlights applications uniquely enabled by mass cytometry in
cancer research.
More information is available on the Fluidigm website: www.fluidigm.com
Mass Cytometry Applications for Cancer Research
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Cell cycle analysis of the human hematopoietic
system by mass cytometry
Reference: Behbehani, G.K., et.al. Cytometry A 81:552-566.
Uniquely CyTOF
 Simultaneous analysis of 36 parameters per sample in human bone marrow
including:
 25 cell surface markers
 8 intracellular functional markers
 Proliferation, DNA content and viability
 Measurement of cell cycle distribution for 34 bone marrow subsets in a single
sample
Mass cytometric analysis of cell cycle distribution in human bone marrow
erythroid cells. Healthy human bone marrow pulsed with 2-iodo-5-deoxyuridine
(IdU) was analyzed using SPADE, an algorithm that clusters cell populations into a
minimum spanning tree according to phenotypic similarities revealed through
usage of 25 cell surface markers. Node color is scaled to the median expression
intensity of CD45 (left) or the indicated markers used to identify erythroid subsets
(right). The erythroid branch of the bone marrow is indicated by the large
rectangle, with subsets shown in the four smaller rectangles. Cluster distribution in
the G1, S, or G2 cell cycle phases is shown by red circles sized according to
relative cellular frequency. Focusing on the erythroid lineage, G1 and S cells were
confined to the erythroblast stage.
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Mass Cytometry Applications for Cancer Research
Simultaneous analysis of 25 cell surface molecules, 8 proteins designating G- and
M-phases and IdU identifying S-phase cells, enabled system-wide mapping of cell
cycle states across hematopoietic differentiation. The information-rich data
provided by mass cytometry paves the way for in-depth investigation of
therapeutic responses at the cell cycle level.
Mapping phenotypic heterogeneity and
progression of cancer cells by mass cytometry
Reference: Amir el, A.D., et.al. Nature biotechnology 31:545-552.
Uniquely CyTOF
 Simultaneous analysis of 33 parameters per sample in healthy and leukemic
human bone marrow including
 31 cell surface markers
 DNA content and viability
 In-depth phenotyping and visualization of cancer cell progression from
diagnosis to relapse
Phenotypic progression of acute myeloid leukemia (AML) cancer cells before
chemotherapy and after disease relapse. Two bone marrow samples from an
AML patient, one collected at diagnosis before chemotherapy, the other collected
at disease relapse, were stained for 31 cell-surface markers then analyzed on a
CyTOF mass cytometer. Data sets were merged and subjected to a nonlinear
unsupervised dimensionality reduction algorithm (viSNE) to collapse high
dimensional single-cell information into a two-dimensional map. The resulting
contour maps (left) highlight cellular portions contributed by the ‘diagnosis’ sample
in purple (upper left) and by the ‘relapse’ sample in red (lower left). Pseudocolored single-cell maps (right) were used to depict mean expression intensities
for the indicated phenotypic markers. The viSNE map reveals pervasive cellular
Flt3 expression unique to the diagnosis sample prior to chemotherapy, and
emerging abnormal immature CD34+ populations displaying hallmarks of lineagecommitted cells at disease relapse.
Mass Cytometry Applications for Cancer Research
5
CyTOF mass cytometry analysis provided detailed characterization of cancer cell
progression from diagnosis to disease relapse on a cell by cell basis. Early and
accurate detection of phenotypically aberrant populations is paramount to
prognosis and correctly assigning treatment decisions. Combining high resolution
examination of cellular heterogeneity and detection of minimal residual disease,
mass cytometry emerges as a powerful method for cancer research.
High-resolution profiling of the baseline diversity in
human NK cells
Reference: Horowitz, A., et al. SciTransl Med 5, 208ra145.
Uniquely CyTOF
 Simultaneous measurement of 38 parameters per cell in blood samples from
monozygotic twins and unrelated human donors including:
 28 NK cell receptors
 8 cell surface phenotyping markers
 DNA content and cell viability
 Use of mass cytometry to reveal a baseline repertoire of over 100,000
phenotypically distinct NK cells shaped by genetic and environmental
determinants in humans
High-dimensional mass cytometry profiling of the NK cell repertoire uncovers a
unique diversity of inhibitory and activating receptor expression patterns.
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Mass Cytometry Applications for Cancer Research
Peripheral blood NK cells from monozygotic twins 5a and 5b (Fig. A) or two
healthy unrelated individuals 003 and 007 (Fig. B) were analyzed by SPADE
through clustering NK cell subsets into a minimum spanning tree according to the
expression of 28 cell surface receptors. Node color is scaled to the median
expression intensity of the activating NKG2D and KIR2SD4 receptors, or the
inhibitory NKG2A and KIR2DL1 receptors. Nodes are sized according to the
relative cell abundance within the entire population of NK cells.
High-resolution phenotyping by mass cytometry uncovered a remarkable breadth
and diversity in human NK cells with over 100,000 phenotypically distinguishable
subsets. By interrogating the diversity of this repertoire in unrelated healthy
individuals and monozygotic twins, inhibitory functions of NK cells were found to
be tightly regulated by host genetics. In striking contrast, the expression and the
combinatorial arrangement of activating receptors is shaped by environmental
cues, thereby ensuring a highly diverse repertoire of NK cells in the face of
infectious challenges and cancer.
Mass Cytometry Applications for Cancer Research
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Bibliography
Han, L., et al. Single-cell mass cytometry reveals intracellular survival/proliferative
signaling in FLT3-ITD-mutated AML stem/progenitor cells. Cytometry A 2015.
Hansmann, L., et al. Mass cytometry analysis shows that a novel memory
phenotype B cell is expanded in multiple myeloma. Cancer Immunol Res 2015.
Chang, Q., et al. Single-cell measurement of the uptake, intratumoral distribution
and cell cycle effects of cisplatin using mass cytometry. Int J Cancer 136 (5): 12021209, 2014.
Di Palma, S., et al. Unraveling cell populations in tumors by single-cell mass
cytometry. Curr Opin Biotechnol 2014.
Fienberg, H.G., et al. Mass Cytometry to Decipher the Mechanism of Nongenetic
Drug Resistance in Cancer. Curr Top Microbiol Immunol 2014.
Giesen, C., et al. Highly multiplexed imaging of tumor tissues with subcellular
resolution by mass cytometry. Nat Methods 2014.
Sachs, Z., et al. NRASG12V oncogene facilitates self-renewal in a murine model of
acute myelogenous leukemia. Blood 2014.
Strauss-Albee, D.M., et al. Coordinated Regulation of NK Receptor Expression in
the Maturing Human Immune System. J Immunol 2014.
Amir el, A.D., et al. viSNE enables visualization of high dimensional single-cell data
and reveals phenotypic heterogeneity of leukemia. Nat Biotechnol 31 (6): 545-552,
2013.
Horowitz, A., et al. Genetic and environmental determinants of human NK cell
diversity revealed by mass cytometry. SciTransl Med 5 (208): 208ra145, 2013.
Tanner, S.D., et al. An introduction to mass cytometry: fundamentals and
applications. Cancer Immunol Immunother 62 (5): 955-965, 2013.
Behbehani, G.K., et al. Single-cell mass cytometry adapted to measurements of
the cell cycle. Cytometry A 81 (7): 552-566, 2012.
Ornatsky, O.I., et al. Study of cell antigens and intracellular DNA by identification of
element-containing labels and metallointercalators using inductively coupled
plasma mass spectrometry. Anal Chem 80 (7): 2539-2547, 2008.
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Mass Cytometry Applications for Cancer Research
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