Name: fabrication of refractive-index-matched devices for biomedical microfluidics
Uploaded: 2018-09-10T14:45:00
Description: Watch this Scientific Journal Video about Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics at JoVE.com

This work was supported by the University of Utah office of the Vice President for Research, as well as by funds in conjunction with grant P30 CA042014 awarded to the Huntsman Cancer Institute and to the CRR Program at the Huntsman Cancer Institute.

1. Fabrication of the Polydimethylsiloxane Negative
<ol>
	<li>Preparation of polydimethylsiloxane
	<ol>
		<li>Measure 18 g of PDMS silicone elastomer and 1.8 g of the curing reagent. Pour the curing reagent into a measuring boat containing the elastomer.</li>
		<li>Mix the elastomer and the curing reagent vigorously for 1 min and put the mixture into a vacuum chamber for 30 min.</li>
		<li>Remove the PDMS from the vacuum, pour 15 g onto the negative using a cookie cutter (radius = 3.8 cm) to keep the P...

<ol>
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MY133-V2000 can be used as an alternative to traditional soft lithography fabrication materials such as PDMS. Previous work has shown that materials with a high index of refraction, such as PDMS, introduce significant artifacts near the channel walls due to the mismatching indices of refraction between the fabrication material and the aqueous solution inside the channel13. MY133-V2000 enables matching the refractive index of the microfluidic device to the aqueous solutions commonly used in biomedi...

The development of microfluidic technology has enabled a wide range of new biomedical techniques that leverage the unique physics of microscopic-scale flows1,2. This includes the diagnostic techniques built on microfluidic platforms that quantify clinically relevant biomarkers, including cell stiffness3, surface markers4, and growth5. By manipulating single cells, microfluidic devices can also be used to measure biomarker heterogeneity, for example as an indicator of malignancy6. The ability to combine microfluidic applications with microscopy has further increased the utility of these platforms by allowing for devices that measure multiple biomarkers simultaneously7.
QPM is a microscopy technique that measures the phase shift as light passes through and interacts with the matter inside transparent samples. The mass of individual cells can be calculated from QPM measurements, by using the known relationship between the refractive index and the biomass density8,9. Previous work has shown that QPM is capable of measuring clinically relevant parameters such as cell growth10,11 and cell mechanical properties via disorder strength12. When combined with microfluidics, QPM can potentially be used to measure cell behavior in a highly controlled environment in vitro. One of the primary challenges facing combining QPM with microfluidics is the high refractive index of most polymers used to construct microfluidic channels via soft lithography13.
An important challenge facing the combination of microfluidics with various microscopy techniques is the mismatch between the refractive index of the device material relative to the refractive index of water14,15. One method to address this is through the use of a low refractive index polymer such as CYTOP16 or MY133-V200013. The latter is a fluorinated ultraviolet (UV)-curable acrylate polymer that has a refractive index similar to water (n = 1.33) and that is compatible with soft lithography techniques, allowing for a smooth integration into many established microfluidic device fabrication workflows. This makes MY133-V2000 not only suitable for microfluidic device fabrication, but also allows it to be readily combined with QPM and other microscopy approaches, to measure cell behavior both at the colony and on a single-cell scale. MY133-V2000 eliminates artifacts due to phase unwrapping by producing little, if any, phase shift as light passes through the water-MY133 interface.
Although eliminating the mismatch in refractive index, one major challenge associated with the devices fabricated from fluorinated polymers, such as MY133-V2000, is the low adherence to other materials such as glass or PDMS. The present work demonstrates the fabrication of an MY133-V2000 microfluidic device using soft lithography. Using O2 plasma to treat the surface of both the channel and the PDMS substrate combined with a custom-fabricated acrylic holder ensures that the device adheres to the substrate, creating a sealed channel. This device is suitable for cell culture and QPM to measure the mass of cells in the channel, which has important applications for measuring the growth of live cells and the intracellular transport of cell biomass, both of which have clinical relevance in diagnostic medicine and drug discovery.

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fabrication of refractive-index-matched devices for biomedical microfluidics

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices can be implemented as part of a robust platform capable of making simultaneous complementary measurements. The primary challenge created by the combination of these two techniques is the mismatch in refractive index between the materials traditionally used to make microfluidic devices and the aqueous solutions typically used in biomedicine. This mismatch can create optical artifacts near the channel or device edges. One solution is to reduce the refractive index of the material used to fabricate the device by using a fluorinated polymer such as MY133-V2000 whose refractive index is similar to that of water (n = 1.33). Here, the construction of a microfluidic device made out of MY133-V2000 using soft lithography techniques is demonstrated, using O2 plasma in conjunction with an acrylic holder to increase the adhesion between the MY133-V2000 fabricated device and the polydimethylsiloxane (PDMS) substrate. The device is then tested by incubating it filled with cell culture media for 24 h to demonstrate the ability of the device to maintain cell culture conditions during the course of a typical imaging experiment. Finally, quantitative phase microscopy (QPM) is used to measure the distribution of mass within the live adherent cells in the microchannel. This way, the increased precision, enabled by fabricating the device from a low index of refraction polymer such as MY133-V2000 in lieu of traditional soft lithography materials such as PDMS, is demonstrated. Overall, this approach for fabricating microfluidic devices can be readily integrated into existing soft lithography workflows in order to reduce optical artifacts and increase measurement precision.

This protocol describes the fabrication of MY133-V2000, a fluorinated polymer with a low refractive index matching that of water. A key feature of this protocol is how to overcome the lack of adhesion that is characteristic of fluorinated polymers by using oxygen plasma and by fabricating the device within an acrylic holder to provide the extra mechanical force required to seal the channel against the PDMS substrate (Figure 1). The low refractive index of the...

Watch this Scientific Journal Video about Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics at JoVE.com

Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics

This protocol describes the fabrication of microfluidic devices from MY133-V2000 to eliminate artifacts that often arise in microchannels due to the mismatching refractive indices between microchannel structures and an aqueous solution. This protocol uses an acrylic holder to compress the encapsulated device, improving adhesion both chemically and mechanically.

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices ...

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Owing to its relativistic low-energy charge carriers, the interaction between graphene and various impurities leads to a wealth of new physics and degrees ...

Fabrication and Operation of Acoustofluidic Devices Supporting Bulk Acoustic Standing Waves for Sheathless Focusing of Particles

Acoustophoresis refers to the displacement of suspended objects in response to directional forces from sound energy. Given that the suspended objects must ...

Fabrication of Fully Solution Processed Inorganic Nanocrystal Photovoltaic Devices

We demonstrate a method for the preparation of fully solution processed inorganic solar cells from a spin and spray coating deposition of nanocrystal ...

Solvent Bonding for Fabrication of PMMA and COP Microfluidic Devices

Thermoplastic microfluidic devices offer many advantages over those made from silicone elastomers, but bonding procedures must be developed for each ...

Scanning Light Scattering Profiler (SLPS) Based Methodology to Quantitatively Evaluate Forward and Backward Light Scattering from Intraocular Lenses

Scanning Light Scattering Profiler SLPS Based Methodology to Quantitatively Evaluate Forward and Backward Light Scattering from Intraocular Lenses

The scanning light scattering profiler (SLSP) methodology has been developed for the full-angle quantitative evaluation of forward and backward light ...

Flow-assisted Dielectrophoresis: A Low Cost Method for the Fabrication of High Performance Solution-processable Nanowire Devices

Flow-assisted dielectrophoresis (DEP) is an efficient self-assembly method for the controllable and reproducible positioning, alignment, and selection of ...

Atmospheric Pressure Fabrication of Large-Sized Single-Layer Rectangular SnSe Flakes

Tin selenide (SnSe) belongs to the family of layered metal chalcogenide materials with a buckled structure like phosphorene, and has shown potential for ...