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…
Disclosures
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 |
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Cite This Article
Label-Free Neutrophil Separation from Tracheal Secretion Using Spiral Microfluidics Channel. J. Vis. Exp. (Pending Publication), e21573, doi: (2023).