A Microfluidic Chip-Based Method for Rapid Neutrophil Chemotaxis Analysis Using Whole Blood

In This Article

Abstract

Source: Yang, K., et al. An All-on-chip Method for Rapid Neutrophil Chemotaxis Analysis Directly from a Drop of Blood. J. Vis. Exp. (124), (2017)

This video demonstrates neutrophil chemotaxis assay using an all-on-chip method. The method allows direct isolation of neutrophils from the whole blood by immunomagnetic negative selection followed by chemotaxis assay on a microfluidic chip.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.

1. Microfluidic device preparation.

Microfluidic Cell Migration Assay Preparation
1. Prepare 50 µg/mL fibronectin solution by diluting 50 µL of stock fibronectin solution (1 mg/mL) to 950 µL Dulbecco's phosphate-buffered saline (DPBS) inside a biosafety cabinet.
2. Prepare the migration medium by mixing 9 mL of Roswell Park Memorial Institute medium (RPMI-1640) and 1 mL of RPMI-1640 with 4% bovine serum albumin (BSA).
3. Remove....

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Results

Microfluidic chip diagram; chemotaxis test; cell isolation; magnetic neutrophil selection process.
Figure 1: Illustration of the all-on-chip method for neutrophil chemotaxis analysis. (A) Illustration of the microfluidic device. The device includes two layers. The first layer (4 µm high) defines the cell docking barrier channel to.......