Frontiers in Immunology (Jul 2021)

Adjacent Cell Marker Lateral Spillover Compensation and Reinforcement for Multiplexed Images

  • Yunhao Bai,
  • Yunhao Bai,
  • Bokai Zhu,
  • Bokai Zhu,
  • Xavier Rovira-Clave,
  • Han Chen,
  • Maxim Markovic,
  • Chi Ngai Chan,
  • Tung-Hung Su,
  • Tung-Hung Su,
  • David R. McIlwain,
  • Jacob D. Estes,
  • Leeat Keren,
  • Leeat Keren,
  • Garry P. Nolan,
  • Sizun Jiang,
  • Sizun Jiang

DOI
https://doi.org/10.3389/fimmu.2021.652631
Journal volume & issue
Vol. 12

Abstract

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Multiplex imaging technologies are now routinely capable of measuring more than 40 antibody-labeled parameters in single cells. However, lateral spillage of signals in densely packed tissues presents an obstacle to the assignment of high-dimensional spatial features to individual cells for accurate cell-type annotation. We devised a method to correct for lateral spillage of cell surface markers between adjacent cells termed REinforcement Dynamic Spillover EliminAtion (REDSEA). The use of REDSEA decreased contaminating signals from neighboring cells. It improved the recovery of marker signals across both isotopic (i.e., Multiplexed Ion Beam Imaging) and immunofluorescent (i.e., Cyclic Immunofluorescence) multiplexed images resulting in a marked improvement in cell-type classification.

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