Label-Free Neutrophil Separation from Tracheal Secretion Using Spiral Microfluidics Channel

Published: August 31, 2023

Abstract

Source: Ryu, H., et al., Label-free Neutrophil Enrichment from Patient-derived Airway Secretion Using Closed-loop Inertial Microfluidics. J. Vis. Exp. (2018).

This video demonstrates separating neutrophils from tracheal secretions using a microfluidic device with spiral channels. The device achieves efficient separation based on cell size by utilizing varying thicknesses and trapezoidal-shaped channels. The interplay between forces results in large neutrophils accumulating near the inner wall while erythrocytes and mucin aggregates remain near the outer wall, ensuring effective neutrophil separation.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Device Fabrication and Soft Lithography NOTE: Standard soft lithography techniques were used to create the polydimethylsiloxane (PDMS) microchannel. Mix the PDMS precursor in a 10:1 ratio of ba…

Representative Results

Figure 1: Spiral microfluidic device and experimental settings. Photograph images of (A) spiral microfluidics and (B) the device assembled with the fluidic adaptor. (C) Schematic diagram of closed-loop separation using spiral inertial microfluidics. By recirculating the particle/cell concentrated stream (IW outlet) back into the original sample reservoir, immune-related cells were concentrated in a small volume while the ba…

Açıklamalar

The authors have nothing to disclose.

Materials

PDMS precursor Dow corning 184 SIL ELAST KIT 3.9KG 10:1 ratio of base and curing agent
VWR gravity convection oven VWR 414005-128 PDMS precursor to be cured in 90 deg.
100mm petri dish VWR 89000-324 Fabrication of PDMS Supporting layer
Harris Uni-core puncher Sigma-aldrich WHAWB100076 2mm diameter or other depending on the tubing size
Air plasma machine Femto Science Cute Surface plasma treatment for PDMS device to bottom base.
2" x 3" glass slide TED PELLA, INC. 2195 To support PDMS device
Masterflex spooled platinum-cured silicone tubing, L/S 14 Cole-Parmer EW-96410-14 Tubing for microfluidics and peristlatic pump
1/16 inch Luer connector, male Harvard apparatus PC2 72-1443 Connector for fluid guide
50mL Falcon tube Corning 21008-940 Sample collection & preparation
Phosphate-Buffered Saline, 1X Without Calcium and Magnesium Corning 45000-446  Buffer solution to dilute sample
Halyard Closed suction Catheter, Elbow, 14F/ channel 4.67mm HALYARD HEALTH 22113 Tracheal seceation suction catheter
0.9% Sterile Normal saline, 10mL pre-filled syringe BD PosiFlush NHRIC: 8290-306547 For tracheal seceation collection from the patients
SecurTainer™ III Specimen Containers, 20mL Simport 1176R36 Sterile sputum (airway secretion) collection container
Syringe with Luer-Lok Tip, 10mL BD BD309604 To pipette homogenize the mucus sample and reach the bottom of sample tube
BD  Blunt Fill Needle, with BD Luer-Lok  Tip BD To pipette homogenize the mucus sample and reach the bottom of sample tube
40µm nylon cell strainer  Falcon 21008-949 To remove large chunk or blood clots, which can block the microfluidics access hole or the channel.
Peristaltic pump (Masterflex L/S Digital Drive) Cole-Parmer HV-07522-30  Operation of microfluidics

Etiketler

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Bu Makaleden Alıntı Yapın
Label-Free Neutrophil Separation from Tracheal Secretion Using Spiral Microfluidics Channel. J. Vis. Exp. (Pending Publication), e21573, doi: (2023).

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