Mass Cytometry Sample Preparation

On the day of sample acquisition, the cells were diluted in 1 ml FACS buffer and washed. The cells were then washed twice in MilliQ water, and then re-suspended at 1 × 10⁶/ml in 1:10 EQ calibration beads (Fluidigm). A minimum number of 500,000 events were then acquired on a mass cytometer (Helios CyTOF, Fluidigm) using CyTOF software (Fluidigm). Following acquisition, all flow cytometry standard (.fcs) files were normalised to bead signal levels using CyTOF software (see [41 (link)] for more details).

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Russo M.A., Fiore N.T., van Vreden C., Bailey D., Santarelli D.M., McGuire H.M., Fazekas de St Groth B, & Austin P.J. (2019). Expansion and activation of distinct central memory T lymphocyte subsets in complex regional pain syndrome. Journal of Neuroinflammation, 16, 63.

Publication 2019

EQ calibration beads Helios CyTOF CyTOF software

Buffer Cells Flow cytometry

Corresponding Organization :

Other organizations : Hunter Genetics, University of Sydney

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Variable analysis

independent variables

Dilution of cells in 1 ml FACS buffer
Washing the cells twice in MilliQ water
Re-suspending the cells at 1 × 10^6/ml in 1:10 EQ calibration beads (Fluidigm)

dependent variables

Events acquired on a mass cytometer (Helios CyTOF, Fluidigm) using CyTOF software

control variables

Minimum number of 500,000 events acquired

controls

EQ calibration beads (Fluidigm) used for normalization of flow cytometry standard (.fcs) files

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