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Preparation of Whole Bone Marrow for Mass Cytometry Analysis of Neutrophil-lineage Cells.

Authors
  • Zhu, Yanfang Peipei1
  • Padgett, Lindsey2
  • Dinh, Huy Q2
  • Marcovecchio, Paola2
  • Wu, Runpei2
  • Hinz, Denise3
  • Kim, Cheryl3
  • Hedrick, Catherine C4
  • 1 Division of Inflammation Biology, La Jolla Institute for Immunology; [email protected]
  • 2 Division of Inflammation Biology, La Jolla Institute for Immunology.
  • 3 Flow Cytometry Core Facility, La Jolla Institute for Immunology.
  • 4 Division of Inflammation Biology, La Jolla Institute for Immunology; [email protected]
Type
Published Article
Journal
Journal of Visualized Experiments
Publisher
MyJoVE Corporation
Publication Date
Jun 19, 2019
Issue
148
Identifiers
DOI: 10.3791/59617
PMID: 31282876
Source
Medline
Language
English
License
Unknown

Abstract

In this article, we present a protocol that is optimized to preserve neutrophil-lineage cells in fresh BM for whole BM CyTOF analysis. We utilized a myeloid-biased 39-antibody CyTOF panel to evaluate the hematopoietic system with a focus on the neutrophil-lineage cells by using this protocol. The CyTOF result was analyzed with an open-resource dimensional reduction algorithm, viSNE, and the data was presented to demonstrate the outcome of this protocol. We have discovered new neutrophil-lineage cell populations based on this protocol. This protocol of fresh whole BM preparation may be used for 1), CyTOF analysis to discover unidentified cell populations from whole BM, 2), investigating whole BM defects for patients with blood disorders such as leukemia, 3), assisting optimization of fluorescence-activated flow cytometry protocols that utilize fresh whole BM.

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