Method Article

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

July 8th, 2025

In This Article

Abstract

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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

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

1. Microfluidic device preparation.

  1. 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

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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.......

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Disclosures

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No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Device fabrication
Mask alignerABMN/A
SpinnerSolitec5000
HotplateVWR11301-022
Plasma cleanerHarrick PlasmaPDC-001
Vacuum dessicatorFisher Scientific08-594-15A
SU-8 2000 thinnerMicrochemSU-8 2000
SU-8 2025 photoresistMicrochemSU-8 2025
(tridecafluoro-1,1,2,2- tetrahydrooctyl) trichlorosilaneGelest78560-45-9
Si waferSilicon, IncLG2065
Glass slidesFisher Scientific12-544-4
Cutting padN/AN/ACustom-made
Punchers
On-chip cell isolation and chemotaxis assay
RPMI 1640Fisher ScientificSH3025502
DPBSFisher ScientificSH3002802
Bovine serum albumin (BSA)Sigma-AldrichSH3057402
FibronectinVWRCACB356008
fMLPSigma-AldrichF3506-10MG
Magnetic disksIndigo Instruments44202-15 mm in diameter, 1 mm thick
FITC-DextranSigma-AldrichFD10S
RhodamineSigma-AldrichR4127-5G
Giemsa stain solutionRowley Biochemical IncG-472-1-8OZ
EasySep Direct Human Neutrophil Isolation KitSTEMCELL Technologies Inc19666
DithiothreitolSigma-AldrichD0632
Nikon Ti-U inverted fluorescent microscopeNikonTi-U
Microscope environmental chamber.InVivo ScientificN/A
CCD cameraNikonDS-Fi1

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Tags

Immunomagnetic SelectionChemoattractant GradientFluorescence MicroscopyCell DockingFibronectin CoatingMigration MediumTemperature Control

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