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Related Experiment Video

Updated: Jun 13, 2025

Quantification and Size-profiling of Extracellular Vesicles Using Tunable Resistive Pulse Sensing

Quantification and Size-profiling of Extracellular Vesicles Using Tunable Resistive Pulse Sensing

Published on: October 19, 2014

Protocol to isolate and quantify large aging neutrophil-derived vesicles.

Alan Y Hsu¹, Qingxiang Huang², Sizhou Feng²

¹Department of Pathology, PhD Program in Immunology, Harvard Medical School, Mass General Brigham, Dana-Farber/Harvard Cancer Center, Boston, MA 02115, USA.

|June 12, 2025

Summary

This summary is machine-generated.

This study details a protocol for isolating large aging neutrophil-derived vesicles (LAND-Vs) from mouse and human samples. The method uses centrifugation and fluorescence-activated cell sorting (FACS) for purification.

Keywords:

Cell Biology Cell isolation Flow Cytometry Immunology

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Last Updated: Jun 13, 2025

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Published on: October 19, 2014

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Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT

Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT

Published on: November 27, 2019

Area of Science:

Cell Biology
Immunology
Extracellular Vesicles

Background:

Neutrophils are key immune cells involved in inflammation and host defense.
Aging neutrophils release large aging neutrophil-derived vesicles (LAND-Vs), which are significant in biological processes.
Understanding LAND-Vs requires effective isolation and purification methods.

Purpose of the Study:

To present a detailed protocol for the identification and purification of murine large aging neutrophil-derived vesicles (LAND-Vs).
To describe the isolation and quantification of human LAND-Vs.
To provide a reproducible method for researchers studying neutrophil extracellular vesicles.

Main Methods:

Isolation of murine LAND-Vs from in vitro neutrophil cultures, bone marrow, blood, and bronchoalveolar lavage fluid (BALF).
Purification of LAND-Vs using differential centrifugation and fluorescence-activated cell sorting (FACS).
Adaptation of the protocol for isolation and quantification of human LAND-Vs from neutrophils and blood.

Main Results:

Successful identification and purification of murine LAND-Vs using the described protocol.
Demonstration of LAND-Vs isolation from various ex vivo sources.
Established methodology for human LAND-Vs isolation and quantification.

Conclusions:

The presented protocol enables efficient isolation and purification of murine and human LAND-Vs.
This method facilitates further research into the biological roles of LAND-Vs.
The protocol is applicable to both in vitro and ex vivo samples, enhancing its utility.