Mass cytometry (MC) measures metal isotope signals from
single
cells and bead samples. Since large numbers of isotopes can be employed
as labels, mass cytometry is a powerful analytical technique for multiparameter
cytometric assays. The calibration protocol in MC is a critical algorithm,
which employs metal-encoded microbeads as an internal standard to
correct the data for instrumental signal drift. The current generation
of commercially available beads carries four lanthanide elements (cerium,
europium, holmium, and lutetium). However, this is not sufficient
to calibrate the full span of detection channels, ranging from yttrium
(89 amu) to bismuth (209 amu), which are now available. To address
this issue we prepared polystyrene microbeads encoded with seven elements
(yttrium, indium, and bismuth in addition to the four lanthanides)
by multistage dispersion polymerization for MC calibration and normalization.
The bead synthesis conditions were optimized to obtain microbeads
that were uniform in size and generated strong MC signal intensities
at similar levels for the eight encoded isotopes. Metal ion leaching
from the beads under storage and application conditions was also examined.
We demonstrated that the precision of normalized MC signals in the
MC detection channels was improved by employing seven-element-encoded
microbeads as a standard.