Development of 1-tube EpiTOF panel

Leipold, Michael;Obermoser, Gerlinde;Maecker, Holden

Description

This CyTOF/mass cytometry dataset summarizes the development of a 1-tube EpiTOF panel.  This compiles Methylation and Acetylation markers into a single panel, rather than 2 panels as in FR-FCM-Z8FS and FR-FCM-Z8GB on FlowRepository.Other differences relative to FR datasets:Addition of more surface markersRevision of gating pathwayReduction of Panel 1 and Panel 2 into a single intracellular panel, due to partial redundancy of certain markers, as determined by the Khatri and Utz labs (https://doi.org/10.1101/2022.01.21.477300 and https://doi.org/10.1016/j.isci.2022.105756)Frozen surface cocktail - more consistent staining between plates/batchesIntracellular cocktail:  roughly half the markers frozen (more consistent staining between plates/batches), roughly half needed to be added as liquid reagents (did not perform well when also frozen)FBS/DMSO -80C freezing of fully stained (including Ir) samples allowed more samples or plates per staining Batch, increasing throughputTrial 11 and 11b finish the development work.  Trial 14, 15, and 15B are the final cocktail prep testing for antibody stocks for a Gates Foundation project.All donors included in testing files are healthy donor PBMCs, processed from LRS chambers from the Stanford Blood Center.  The files were acquired on a Helios mass cytometer (Standard BioTools), normalized using the CyTOF software and the included EQ4 beads (then concatenated if necessary), then debarcoded using the CyTOF software (20plex Pd barcoding kit as in the included staining protocol).  Only the Normalized (concatenated) Debarcoded files are included in this dataset. Grant ID:  INV-008378Funding Source:  Bill and Melinda Gates Foundation We also acknowledge the Purvesh Khatri Lab and PJ Utz Lab (both Stanford University) for initial development.

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Metrics

Dataset Index

0.5

FAIR Score

73%

Citations

0

Mentions

0

Metrics Over Time

Publication Details

DOI

Publisher

Zenodo

License

Creative Commons Attribution 4.0 International

Assigned Domain

Subfield

Rheumatology

Field

Medicine

Domain

Health Sciences

Confidence Score

34%

Source

Scholar Data Model

Keywords

CyTOFmass cytometry

Normalization Factors

FT

51.92

CTw

1.00

MTw

1.00