Microfluidic Chips Controlled with Elastomeric Microvalve Arrays

Summary

We demonstrate protocols for manufacturing and automating elastomeric polydimethylsiloxane (PDMS)-based microvalve arrays that need no extra energy to close and feature photolithographically defined precise volumes. A parallel subnanoliter-volume mixer and an integrated microfluidic perfusion system are presented.

Abstract

Miniaturized microfluidic systems provide simple and effective solutions for low-cost point-of-care diagnostics and high-throughput biomedical assays. Robust flow control and precise fluidic volumes are two critical requirements for these applications. We have developed microfluidic chips featuring elastomeric polydimethylsiloxane (PDMS) microvalve arrays that: 1) need no extra energy source to close the fluidic path, hence the loaded device is highly portable; and 2) allow for microfabricating deep (up to 1 mm) channels with vertical sidewalls and resulting in very precise features.

The PDMS microvalves-based devices consist of three layers: a fluidic layer containing fluidic paths and microchambers of various sizes, a control layer containing the microchannels necessary to actuate the fluidic path with microvalves, and a middle thin PDMS membrane that is bound to the control layer. Fluidic layer and control layers are made by replica molding of PDMS from SU-8 photoresist masters, and the thin PDMS membrane is made by spinning PDMS at specified heights. The control layer is bonded to the thin PDMS membrane after oxygen activation of both, and then assembled with the fluidic layer. The microvalves are closed at rest and can be opened by applying negative pressure (e.g., house vacuum). Microvalve closure and opening are automated via solenoid valves controlled by computer software.

Here, we demonstrate two microvalve-based microfluidic chips for two different applications. The first chip allows for storing and mixing precise sub-nanoliter volumes of aqueous solutions at various mixing ratios. The second chip allows for computer-controlled perfusion of microfluidic cell cultures.

The devices are easy to fabricate and simple to control. Due to the biocompatibility of PDMS, these microchips could have broad applications in miniaturized diagnostic assays as well as basic cell biology studies.

Protocol

Microfluidic device design using CorelDraw or AutoCAD software

Principle of PDMS microvalves-based devices: The devices consist of three layers: a fluidic layer containing microchambers of various sizes, a "control layer" containing the microchannels necessary to actuate the fluidic path with microvalves, and a middle thin PDMS membrane that is bound to the control layer. At rest, due to the compliance and hydrophobicity of PDMS, the membrane seals (reversibly) against its seat, therefore the chambers remain isolated from each other without energy input. Valves can be opened by applying negative pressure (e.g., house vacuum),....

Discussion

Main advantages of our microvalve design:

1. No extra energy source is required to close the fluidic path, hence the loaded device is highly portable; and
2. The device can be built by PDMS replicas from photolithographically-patterned SU-8 molds, allowing for microfabricating deep (up to 1 mm) channels with vertical sidewalls (i.e. the height of the features can be specified independently of their width) and resulting in very precise features.

Advantages of the par.......

Materials

Material Name	Type	Company	Catalogue Number	Comment
Name	Company	Catalog Number	Comments
Clean silicon wafers	Supplies	Silicon Sense Inc.	3P0110TEST	3-inch diameter, P/Boron
"Master" wafers containing SU-8 patterns	Supplies			Fabricated in house using standard photolithography procedures
Desiccators (2)	Equipment	VWR	24987-048	One for silanization, one for PDMS de-bubbling.
Balance	Equipment	OHAUS Corp.	SC6010
Oven	Equipment	Sheldon Mfg. Inc.	1330GM
MiniVortexer	Equipment	VWR	58816-121
Spinner	Equipment	Headway Research Inc.	PWM32
Plasma etcher	Equipment	Plasmatic Systems Inc.	Plasma Preen II-973
Hot Plate	Equipment	Torre Pines Scientific	HP30A
Stereoscope	Microscope	Nikon	TMZ1500
CCD camera	Equipment	Diagnostic Instruments	SPOT RT
Solenoid valves	Equipment	Lee Company	LHDA0511111H
Data acquisition board	Hardware	National Instruments	PCI 6025E, CB-50LP
LabView	Software	National Instruments	Version 8.0
Tridecafluoro-1,1,2,2,-tetrahydrooctyl)-1-trichlorosilane	Reagent	United Chemical Technologies	T2492	Silanization must be done in a chemical fume hood.
PDMS prepolymer and crosslinker	Reagent	Dow-Corning	Sylgard 184
Hexane	Reagent	EMD	HX0295-6
Color Dyes	Reagent	Spectrum Chemical Mfg. Corp.	FD&C 110, 135, 150	Blue #1, Yellow #5, Red #3.
3 ml disposable transfer pipets	Supplies	Fisher Scientific	13-711-20
Kimwipes	Supplies	Kimberly-Clark	34155
Weighing boats	Supplies	VWR	12577-027
Tongue depressor	Supplies	Fisher Scientific	11-700-555
P100 dishes	Supplies	Fisher Scientific	08-772E
Silicone tubing (1.14 mm inner diameter (I.D.))	Supplies	Cole-Palmer Instrument Co.	07625-30
Tygon tubing (O.D. 1/16 in; I.D. 1/32 in)	Supplies	Cole-Palmer Instrument Co.	06418-02
Duco Cement	Supplies	Devcon	6245
Razor blade	Tools	VWR	55411-050
Needles	Tools	Fisher Scientific	0053482 (25 Gauge)
#5 Forceps	Tools	Fine Science Tools	11251-20
50 ml centrifuge tube	Supplies	Fisher Scientific	05-526B
Seal wrap film	Supplies	AEP Industries Inc.	0153877
1.5 ml microcentrifuge tubes	Supplies	Fisher Scientific	05-406-16
15 ml centrifuge tubes	Supplies	BD Falcon	352097
Purple nitrile power-free gloves	Supplies	VWR	40101-348
1.2 mm Harris biopsy punch	Tools	Ted Pella, Inc.	15074