AutoSpectral: 40-colour ID7000 dataset

Published: 29 October 2025| Version 2 | DOI: 10.17632/2kvc98hc65.2
Contributors:
Oliver Burton, Leoni Buecken, Leana De Vuyst,
,
,

Description

Healthy human PBMC and buffy coat samples, cryopreserved and stained with a 40-colour flow cytometry panel. Acquired on a 5-laser Sony ID7000 spectral instrument. Unmixed using WLSM on the Sony software or using AutoSpectral. WLSM: Sony software unmixed using weighted least squares WLS: AutoSpectral unmixed using weighted least squares scAF: AutoSpectral unmixed using weighted least squares and per-cell (single cell) autofluorescence extraction perCellFluor: AutoSpectral unmixed using weighted least squares, per-cell autofluorescence extraction and per-cell fluorophore spectral optimization. Slow = exact recomputation of the unmixing matrix per variant, Medium = rank-one updating, Fast = approximation using the difference in spectrum between variants and the abundance of the endmember (fluorophore).

Files

Steps to reproduce

Contact Stephanie Humblet-Baron for the staining protocol. For unmixing, see instructions on the GitHub repository for AutoSpectral.

Institutions

  • University of Cambridge
  • Katholieke Universiteit Leuven

Categories

Flow Cytometry, Spectral Unmixing

Funders

Licence