Ultralocalized thermal reactions in subnanoliter droplets-in-air

Miniaturized laboratory-on-chip systems promise rapid, sensitive, and multiplexed detection of biological samples for medical diagnostics, drug discovery, and high-throughput screening. Within miniaturized laboratory-on-chips, static and dynamic droplets of fluids in different immiscible media have been used as individual vessels to perform biochemical reactions and confine the products. Approaches to perform localized heating of these individual subnanoliter droplets can allow for new applications that require parallel, time-, and space-multiplex reactions on a single integrated circuit. Our method positions droplets on an array of individual silicon microwave heaters on chip to precisely control the temperature of droplets-in-air, allowing us to perform biochemical reactions, including DNA melting and detection of single base mismatches. We also demonstrate that ssDNA probe molecules can be placed on heaters in solution, dried, and then rehydrated by ssDNA target molecules in droplets for hybridization and detection. This platform enables many applications in droplets including hybridization of low copy number DNA molecules, lysing of single cells, interrogation of ligand–receptor interactions, and rapid temperature cycling for amplification of DNA molecules.

[1]  S Tomić,et al.  Dielectric relaxation of DNA aqueous solutions. , 2007, Physical review. E, Statistical, nonlinear, and soft matter physics.

[2]  Kenji Yasuda,et al.  Development of 1480 nm Photothermal High-Speed Real-Time Polymerase Chain Reaction System for Rapid Nucleotide Recognition , 2008 .

[3]  Scott L Diamond,et al.  Printing chemical libraries on microarrays for fluid phase nanoliter reactions , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Gwo-Bin Lee,et al.  Integrated microfluidic systems for cell lysis, mixing/pumping and DNA amplification , 2005 .

[5]  Helen Song,et al.  Reactions in droplets in microfluidic channels. , 2006, Angewandte Chemie.

[6]  J P Landers,et al.  Polymerase chain reaction in polymeric microchips: DNA amplification in less than 240 seconds. , 2001, Analytical biochemistry.

[7]  Rashid Bashir,et al.  Nanoscale thickness double-gated field effect silicon sensors for sensitive pH detection in fluid , 2008 .

[8]  P. Sorger,et al.  Electronic detection of DNA by its intrinsic molecular charge , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Oguz H. Elibol,et al.  Localized heating on silicon field effect transistors: device fabrication and temperature measurements in fluid. , 2009, Lab on a chip.

[10]  J. Wölcke,et al.  Miniaturized HTS technologies - uHTS. , 2001, Drug discovery today.

[11]  Darwin R. Reyes,et al.  Microwave dielectric heating of fluids in an integrated microfluidic device , 2007 .

[12]  Bernard P. Puc,et al.  An integrated semiconductor device enabling non-optical genome sequencing , 2011, Nature.

[13]  B. Müller,et al.  Drug Liposome Partitioning as a Tool for the Prediction of Human Passive Intestinal Absorption , 1999, Pharmaceutical Research.

[14]  T. Litovitz,et al.  On the mechanism of dielectric relaxation in aqueous DNA solutions , 1991, Biopolymers.

[15]  Oguz H. Elibol,et al.  High-k dielectric Al2O3 nanowire and nanoplate field effect sensors for improved pH sensing , 2011, Biomedical microdevices.

[16]  T. Ha,et al.  Probing single-stranded DNA conformational flexibility using fluorescence spectroscopy. , 2004, Biophysical journal.

[17]  M. Bittner,et al.  Expression profiling using cDNA microarrays , 1999, Nature Genetics.

[18]  Luigi Colombo,et al.  Effect of nitrogen on band alignment in HfSiON gate dielectrics , 2005 .

[19]  M Graf,et al.  Micro hot plate-based sensor array system for the detection of environmentally relevant gases. , 2006, Analytical chemistry.

[20]  D. Stuerga.  Microwave‐Material Interactions and Dielectric Properties, Key Ingredients for Mastery of Chemical Microwave Processes , 2008 .

[21]  L. R. van den Doel,et al.  Monitoring enzymatic reactions in nanolitre wells , 2003, Journal of microscopy.

[22]  Rashid Bashir,et al.  Ultra-localized single cell electroporation using silicon nanowires. , 2013, Lab on a chip.

[23]  Inkyu Park,et al.  Selective surface functionalization of silicon nanowires via nanoscale joule heating. , 2007, Nano letters.

[24]  David Issadore,et al.  Microwave dielectric heating of drops in microfluidic devices. , 2009, Lab on a chip.

[25]  Rashid Bashir,et al.  Electrical detection of dsDNA and polymerase chain reaction amplification , 2011, Biomedical microdevices.

[26]  Hiroyuki Fujita,et al.  Microfabricated arrays of femtoliter chambers allow single molecule enzymology , 2005, Nature Biotechnology.

[27]  Siarhei Vishniakou,et al.  Petri dish PCR: laser-heated reactions in nanoliter droplet arrays. , 2009, Lab on a chip.

[28]  Axel Scherer,et al.  Thermal management in microfluidics using micro-Peltier junctions , 2005 .

[29]  C. Gabriel,et al.  Dielectric behavior of DNA solution at radio and microwave frequencies (at 20 degrees C). , 1984, Biophysical journal.

[30]  A. Brookes,et al.  Dynamic allele-specific hybridization. A new method for scoring single nucleotide polymorphisms. , 1999, Nature biotechnology.

[31]  Magnus Jobs,et al.  Dynamic allele-specific hybridization , 1999, Nature Biotechnology.

[32]  Yuzuru Takamura,et al.  On-chip nanoliter-volume multiplex TaqMan polymerase chain reaction from a single copy based on counting fluorescence released microchambers. , 2004, Analytical chemistry.

[33]  Oguz H. Elibol,et al.  Silicon field effect transistors as dual-use sensor-heater hybrids. , 2011, Analytical chemistry.

[34]  Sheng-Ce Tao,et al.  A universal multiplex PCR strategy for 100-plex amplification using a hydrophobically patterned microarray. , 2011, Lab on a chip.

[35]  Oguz H. Elibol,et al.  Surface Immobilizable Chelator for Label-free Electrical Detection of Pyrophosphatew Chemcomm , 2022 .

[36]  Carl T Wittwer,et al.  High-resolution DNA melting analysis for simple and efficient molecular diagnostics. , 2007, Pharmacogenomics.

[37]  Y. Murakami,et al.  High-throughput PCR in silicon based microchamber array. , 2001, Biosensors & bioelectronics.

[38]  David J. Mooney,et al.  Label-free biomarker detection from whole blood , 2009, 2010 10th IEEE International Conference on Solid-State and Integrated Circuit Technology.

[39]  S. Yagihara,et al.  Dynamics and structure of water bound to DNA , 1988 .