Mon Aug 06, 2018 3:47 pm
2018-Willis et al-Cytometry A
"Tellurium-based mass cytometry barcode for live and fixed cells"
Willis LM, Park H, Watson MWL, Majonis D, Watson JL, Nitz M
Cytometry A. 2018
http://dx.doi.org/10.1002/cyto.a.23495
- no dataset accession number given
- nonpolar Te-based molecule
- See Fig 4 for live-cell BC
- Jurkat IC50 13uM, staining at 2uM or less
"The results in Figure 4C,D indicate that the 130 isotopologue has a low level of leaching to other cells. This issue may be due to an undetected low level chemical impurity in the 130TeMal sample. The 130Te leaching appears stronger than it is due to the mass sensitivity curve of the CyTOF‐2TM instrument. A solution standard of natural tellurium was analyzed in solution mode on the CyTOF‐2TM, and after taking into account the natural isotopic abundance, we found that the 126, 128, and 130 channels were 12, 41, and 71% more sensitive when compared to the 124 channel. When we subtract the reported isotopic impurities (of the starting metals) but do not account for mass sensitivity of the instrument, the TeMal‐stained cells in Figure 4B have ∼5.5, 4.5, and 2% impurity of 130Te (for the 124Te, 126Te, and 128Te‐stained cells, respectively); however, when mass sensitivity is accounted for, the impurities are ∼3.5, 3.5, and 1.5%, respectively. "
"We also demonstrated that TeMal is compatible with the Callisto system for automated microscale cell culture. Callisto is a fully environmentally controlled system facilitates combinatorial dosing of adherent cells cultured in 32 microfluidic chambers, each being individually treated and addressable. The current barcoding kit reagents are incompatible with Callisto because the barcoding reagent reacts with the protein pre‐coated on the microfluidic channels and the Matrigel used to support the iPS cells. Similar difficulties would be seen with other proposed cell labeling strategies such as OsO4 and RuO4 18. TeMal is sulfhydryl‐reactive and therefore does not interact with pre‐coated protein and reaches microchambers at known concentration. TeMal is clearly compatible with the Callisto system and may facilitate multi‐parameter proteomic analysis of precious primary tissue samples."
Willis LM, Park H, Watson MWL, Majonis D, Watson JL, Nitz M
Cytometry A. 2018
http://dx.doi.org/10.1002/cyto.a.23495
- no dataset accession number given
- nonpolar Te-based molecule
- See Fig 4 for live-cell BC
- Jurkat IC50 13uM, staining at 2uM or less
"The results in Figure 4C,D indicate that the 130 isotopologue has a low level of leaching to other cells. This issue may be due to an undetected low level chemical impurity in the 130TeMal sample. The 130Te leaching appears stronger than it is due to the mass sensitivity curve of the CyTOF‐2TM instrument. A solution standard of natural tellurium was analyzed in solution mode on the CyTOF‐2TM, and after taking into account the natural isotopic abundance, we found that the 126, 128, and 130 channels were 12, 41, and 71% more sensitive when compared to the 124 channel. When we subtract the reported isotopic impurities (of the starting metals) but do not account for mass sensitivity of the instrument, the TeMal‐stained cells in Figure 4B have ∼5.5, 4.5, and 2% impurity of 130Te (for the 124Te, 126Te, and 128Te‐stained cells, respectively); however, when mass sensitivity is accounted for, the impurities are ∼3.5, 3.5, and 1.5%, respectively. "
"We also demonstrated that TeMal is compatible with the Callisto system for automated microscale cell culture. Callisto is a fully environmentally controlled system facilitates combinatorial dosing of adherent cells cultured in 32 microfluidic chambers, each being individually treated and addressable. The current barcoding kit reagents are incompatible with Callisto because the barcoding reagent reacts with the protein pre‐coated on the microfluidic channels and the Matrigel used to support the iPS cells. Similar difficulties would be seen with other proposed cell labeling strategies such as OsO4 and RuO4 18. TeMal is sulfhydryl‐reactive and therefore does not interact with pre‐coated protein and reaches microchambers at known concentration. TeMal is clearly compatible with the Callisto system and may facilitate multi‐parameter proteomic analysis of precious primary tissue samples."