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Updated: Sep 14, 2025

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

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

Published on: June 19, 2019

Optimization of Cytometry by Time-of-Flight Staining for Peripheral Blood and Bone Marrow Samples.

Yosuke Susuki¹, Issei Shinohara¹, Masatoshi Murayama¹

¹Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, California, USA.

Tissue Engineering. Part C, Methods

|July 21, 2025

Summary

This study optimizes Cytometry by time-of-flight (CyTOF) staining for immune profiling. The refined protocol improves signal quality for key markers in blood and bone marrow samples, advancing translational research.

Keywords:

CyTOF bone marrow aspiration concentrate osteonecrosis of the femoral head peripheral blood mononuclear cells

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Last Updated: Sep 14, 2025

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Database-guided Flow-cytometry for Evaluation of Bone Marrow Myeloid Cell Maturation

Database-guided Flow-cytometry for Evaluation of Bone Marrow Myeloid Cell Maturation

Published on: November 3, 2018

Area of Science:

Immunology
Biotechnology
Translational Research

Background:

Cytometry by time-of-flight (CyTOF) is crucial for immune profiling in translational research.
Existing CyTOF protocols face challenges like signal variability, nonspecific binding, and antibody incompatibility, impacting data quality.

Purpose of the Study:

To present an optimized CyTOF staining protocol for human peripheral blood mononuclear cells and bone marrow samples.
To address common challenges in CyTOF data acquisition and improve signal consistency and clarity.

Main Methods:

Refined antibody conjugation using polymer X8.
Optimized saponin use and fixation protocols for sample preparation.
Application to human peripheral blood mononuclear cells and bone marrow aspiration concentrate.

Main Results:

Preliminary data show improved staining for key immune markers, including CD14, CD16, and CD19.
Enhanced signal consistency and clarity were observed in the processed samples.
The optimized protocol addresses challenges of signal variability and nonspecific binding.

Conclusions:

The optimized CyTOF staining protocol enhances data quality for immune profiling.
This advancement is particularly relevant for orthopaedic research and understanding immune responses in diseases like osteonecrosis of the femoral head.