Microfluidic Thermal Cycler with Electrowetting

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Presentation transcript:

Microfluidic Thermal Cycler with Electrowetting Quintin T. Davis With Dr. Stanislaw Legowski

Intent of Project Goal: Investigate droplet microfluidics as a method for manipulating 10 microliter scale volumes utilizing electrowetting for the completion of thermal cycling protocols. Application: The Polymerase Chain Reaction

Background Overview Background: Polymerase Chain Reactions Electrowetting: The relationship between surface tension and electric field Application to lab protocols

Background PCR: method of amplifying DNA segments by repeated melting, reannealing and copying. Ubiquitous in medicine and biology Typical heating / cooling rates: 3 / 2 °C / s-1

PCR Denaturation: 94C Reannealling: ~(Tm – 4C ) Extension 72C Taq polymerase processes ~300 nt*s-1

Electrowetting First described by Gabriel Lippmann in 1875 [2]. Now being investigated for automation and downscaling of lab protocols.

Project Overview Requires experiment to determine unknowns: surface hydrophobicity, droplet actuation and heating. Project components: Voltage source Switching array Temperature sensing Heating

Surface wettability Wettability is the tendency of a liquid to spread out over a surface. Contact angles are a method of measured the wettability of a surface, e.g:

Contact angle is related to electric field by the Young-Lippmann eqn. Young equation: Lippmann-Young equation relates contact angle to electric potential, dielectric strength and thickness: (Mugele and Baret 2005)

Solder mask thickness from (Paik 2005)

Voltage Source Voltage depends on transformer turns (from data sheets)

Switching STS1HNK60, N-channel MOSFET with VDSMax = 600V, logic-level input with Threshold 2.25 < VGS < 3.0 74HC154 Active-low demultiplexer (VOH >= 0.95 * VDD) Used 3 input pins for 23 = 8 switch control. Add references Print out relevant datasheets and papers Take picture of SMBus communication and add slide

Radiation cooling obviates the need for active cooling J = T4A Stefan-Boltzmann equation

Non-Contact Infrared Thermometry uses SMBus Serial Communication Protocol Derivative of I2C uses one data line (SDA) and one clock line. MLX90614 IR thermometers are individually addressable by unique 7-bit addresses.

SMBus Communication cont’d

Temperature sensing and heating platform An large scale model for a microfluidic thermal reaction chamber

Thermal cycling test results Boiling point at 7200 ft: ~92.7 °C

Electrowetting

Electrowetting Cont’d

Conclusions Inconsistent droplet actuation at ~300 V. Teflon is an undesirable coating for biological laboratory applications due to cost, licensing, lack of robustness and a lack of adherence to many surfaces, including the solder mask used in this project. For low-volume throughput applications such as a typical MOLB lab or rural medical diagnostics, electrowetting represents an uneccessary jump in complexity for PCR.

Acknowledgements Dr. Stanislaw Legowski Dr. Naomi L. Ward Thanks to: Dr. Shawna McBride, Michelle Turner, Dr. Cameron Wright, Dr. Jon Pikal, Dr. Steve Barret This work funded by Wyoming NASA Space Grant Consortium, NASA Grant #NNG05G165H

[1]Paik P, Pamula V, Pollack M, Chakrabarty K [1]Paik P, Pamula V, Pollack M, Chakrabarty K. Coplanar Digital Microfluidics Using Standard Printed Circuit Board Processes. 2005. Conf. on Miniaturized Systems for Chem. And Life Sciences. 9. [2]Mugele F, Baret J-C. Electrowetting: from Basics to Applications. 2005. J. Phys. Condens. Matter. 17. R705-R774. [3]Kim H, Dixit S, Green C, Faris G. Nanodroplet real-time PCR system with laser assisted heating. 2009. Optics Express. 17(1). 218-228. [4]Neuzil P, Zhang C, Pipper J, Oh S, Zhuo L. Ultra fast miniaturized real-time PCR: 40 cycles in less than six minutes. 2006. Nucleic Acids Res. 34(11). e77-e86.