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

Published: January 31, 2024

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 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. Prepare the mig…

Representative Results

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 trap the cells beside the gradient channel. The second layer (60 µm high) defines the gradient generating channel, the port and channel fo…

Disclosures

The authors have nothing to disclose.

Materials

Device fabrication
Mask aligner ABM N/A
Spinner Solitec 5000
Hotplate VWR 11301-022
Plasma cleaner Harrick Plasma PDC-001
Vacuum dessicator Fisher Scientific 08-594-15A
SU-8 2000 thinner Microchem SU-8 2000
SU-8 2025 photoresist Microchem SU-8 2025
(tridecafluoro-1,1,2,2- tetrahydrooctyl) trichlorosilane Gelest 78560-45-9
Si wafer Silicon, Inc LG2065
Glass slides Fisher Scientific 12-544-4
Cutting pad N/A N/A Custom-made
Punchers
On-chip cell isolation and chemotaxis assay
RPMI 1640 Fisher Scientific SH3025502
DPBS Fisher Scientific SH3002802
Bovine serum albumin (BSA) Sigma-Aldrich SH3057402
Fibronectin VWR CACB356008
fMLP Sigma-Aldrich F3506-10MG
Magnetic disks Indigo Instruments 44202-1 5 mm in diameter, 1 mm thick
FITC-Dextran Sigma-Aldrich FD10S
Rhodamine Sigma-Aldrich R4127-5G
Giemsa stain solution Rowley Biochemical Inc G-472-1-8OZ
EasySep Direct Human Neutrophil Isolation Kit STEMCELL Technologies Inc 19666
Dithiothreitol Sigma-Aldrich D0632
Nikon Ti-U inverted fluorescent microscope Nikon Ti-U
Microscope environmental chamber. InVivo Scientific N/A
CCD camera Nikon DS-Fi1

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Cite This Article
A Microfluidic Chip-Based Method for Rapid Neutrophil Chemotaxis Analysis Using Whole Blood. J. Vis. Exp. (Pending Publication), e21903, doi: (2024).

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