Miniaturization in bioanalysis Microfluidics – Introduction/history Applications - mass spectrometry Patents Terry, S.C., Jerman, J.H., Angell, J.B. IEEE Transactions on Electron Devices, 1979, 26, 1880-86. A Gas Chromatographic Air Analyzer Fabricated on a Silicon Wafer Picture by Wyatt Vreeland, NIST There really is nothing new under the Sun Prototype analog pneumatic computer to operate in a nuclear attack NBS (now NIST) 1950’s https://www.ncbi.nlm.nih.gov/pubmed?term=Microfluidic%5BTitle%5D Incidence of the word “MICROFLUIDIC” in PubMed (as of August 2018) 1997 2000 2005 2010 2015 2018 Stephen Quake, Dept. Bioengineering, Stanford University, http://thebigone.stanford.edu/index.html Technology Products Consequences Microelectronics Microfluidics ? Microfluidics? Speed of analysis Space saving Cost cutting Mass production Control of electric current control of fluid flows 1. Expose 2. develop and etch mask photoresist protecting layer (Cr, Au) Substrate (glass, silicon, ...) 3. Remove the protection layer 4. Close the structure (thermal bonding) Photolithography Microfabrication technology Micromilling 10 mm Optical lithography 200 nm e/ion beam lithography Multiple exposure techniques 10 nm Etching (resist dependent) ~ nm Replication (mass production) 10’s nm Injection molding Hot embossing Casting • Substrates up to 100 x 100 mm2 • Structures down to 1 μm • Address grid down to 40 nm • Standard or UV laser source mPG 101 Tabletop Laser Pattern Generator System Integration Spatial flow focusing Sankha Shuvra Das, Binay Kumar Patawari, P.K. Patowari, S Halder 5th International & 26th AIMTDR 2014, December 12th–14th, 2014, IIT Guwahati, Assam, India 236-1 Computational Analysis for Mixing of Fluids Flowing through MicroChannels Diffusion limited mixing Capillary force filling www.gyros.com Exact volume metering on the nl level sample Defined volume (nl) Hydrophobic patch http://www.dolomite-microfluidics.com/ Droplet generation in nl-pl volumes Seth Fraden et al., J. AM. CHEM. SOC. 2007, 129, 8825-8835. Benefits and Issues Size - space saving Low reagent/sample consumption Smaller size – faster analysis Microchannel junctions without dead volume Parallel systems for high throughput Disposable parts - point-of-care devices BUT Scaling issues Fabrication limitations Surface chemistry Concentration limits of detection Phenomena unimportant on the macro scale may dominate Small volume problem 2.15 mm => 10 ml ~ 10-15 M 1 mm => 1 ml ~ 10-14 M 0.1 mm => 1 nl ~ 10-11 M 0.001 mm => 1 fl ~ 10-5 M Example: LOD = 1000 molecules MICROFABRICATED DEVICES * Sensors - accelerometers, glucose monitors, ... * Genomics - next gen sequencing * Proteomics - sample processing separation Yining Shi, Peter C. Simpson, James R. Scherer, David Wexler, Christine Skibola, Martyn T. Smith, and Richard A. Mathies. Anal. Chem. 1999, 71, 5354-5361 Radial Capillary Array Electrophoresis Microplate and Scanner for High-Performance Nucleic Acid Analysis. Electrospray Flat surface - droplet Capillary tip Etching, pulling, … Grinding Original capillary ESI tip (micro)fabrication ? ESI exit spray fluid BGE waste sample nebulizer gas 10 mm Zhang, B. Liu, H. Karger, B. L. Foret, F. Anal. Chem., 1999, 71, 3258-3264. CE Microdevice with a Pneumatic Nebulizer 0 2.0 4.0 Time (min) 0 20 40 60 80 100 RelativeAbundance m/z 400 1200 591.6 1181.7 Angiotensin I des-AspI 450.5 899.5 630.6 775.4 m/z 400 1200 m/z 400 1200 m/z 400 1200 Angiotensin III 1-7 Angiotensin I Bullfrog Angiotensin III 3-8 Base peak 441.1 880.8 m/z 400 1200 Angiotensinogen 1-14 Porcine 1 2 3 4 5 CE Microdevice with a Pneumatic Nebulizer 1. 2. 3. 4. 5. Zhang, B. Liu, H. Karger, B. L. Foret, F. Anal. Chem., 1999, 71, 3258-3264. 100 mm Integrated Au tips ESI tip fabrication www.advion.com DRIE in silicone Plasma etched in polyimide www.diagnoswiss.com www.phoenix-st.com www.agilent.com Injection molding in polypropylene Applications Commercialization Microfluidic Chip for Peptide Analysis with an Integrated HPLC Column, Sample Enrichment Column, and Nanoelectrospray Tip H. Yin, K. Killeen, R. Brennen, D. Sobek, M. Werlich, T. van de Goor Anal. Chem. 2005, 77, 527-533 Polyimide HPLC-chip, integrating an enrichment column, frits, a laser ablated ESI tip and trapazoidal separation column www.agilent.com Three LC columns – length 130 mm Each segment individually packed. BSA digest separated with a 30min gradient on a 2 column segmented chip, packed with 3.5μm particles Segmented column HPLC/chip www.agilent.com TRIZAIC nanoTile - Waters Enolase digest 70 fmol, 2 mm particles nanoACQUITY TRIZAIC nanoTile Green tape Al2O3–MgO–SiO2 glass particles mixed with organic binders and solvents to form glass ceramic Product Description 951 Green Tape is a low-temperature cofired ceramic tape. The 951 system comprises a complete cofireable family of Au and Ag metallizations, buried passives, and encapsulants. 951 is available in multiple thicknesses for use as an insulating layer in: •Multichip modules •Single chip packages •Ceramic printed wiring boards •RF modules http://www.dupont.com/mcm Cut Ceramic Material Fired Wafer Ceramic Microfluidic Fabrication Final Diced Part Microfabricated Monolith Columns for Liquid Chromatography Sculpting Supports for Liquid Chromatography Fred E. Regnier J. High Resol. Chromatogr. 2000, 23, (1) 19–26 A microfabricated micropillar liquid chromatographic chip Monolithically integrated with an electrospray ionization tip Lauri Sainiemi, Teemu Nissilä, Risto Kostiainen, Sami Franssila and Raimo A. Ketola Lab Chip, 2012, 12, 325 https://www.pharmafluidics.com/ http://www.chromatographyonline.com/evaluation-micro-pillar-array-columns-pac-combined-high-resolution-mass-spectrometry-lipidomics?pageID=5 Miniaturized (microfabricated) mass spectrometers? Applications of Miniaturized MS Instruments • trace explosive detection and airport security • space exploration • environmental monitoring • point-ofcare medical applications Miniature Mass Spectrometer Systems based on a Microengineered Quadrupole Filter A. Malcolm, S.Wright, R. R. A. Syms, N. Dash, M.-A. Schwab, A. Finlay Anal. Chem. 2010, 82, 1751–1758 Mass Analyzer ionchip® quadrupole mass spectrometer Mass Range m/z 50-800 with ionchip®150 Mass Accuracy ± m/z 0.3 in full scan Mass Resolution m/z 0.7±0.1 FWHM Sensitivity 10pg of reserpine in SIM mode S/N ratio of 10:1 (RMS) www.microsaic.com Advion expression Compact Mass Spectrometer www.advion.com http://908devices.com/ Microscale Ion Trap Internal glow discharge ionization Mass Range 15-450 amu Mass Resolution 1 amu Automated CE-MS on a glass chip What is a patent Invention disclosure Does it make sense to patent? Patent search Resources Patent? Patent! What Is a Patent? A patent for an invention is the grant of a property right to the inventor, issued by the United States Patent and Trademark Office. Generally, the term of a new patent is 20 years from the date on which the application for the patent was filed in the United States or, in special cases, from the date an earlier related application was filed, subject to the payment of maintenance fees. 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Does not fall under the laws of nature, natural phenomena or abstract ideas 2. Utility requirement - invention must be useful in association with machines, human-made products, compositions of matter or processing methods 3. Novelty the idea must not be presented to the public before the filing 4. Nonobviousness – it must be unrecognizable to a skilled person in the field of invention 5. Clarity of the description included in the application Patent je zákonná ochrana vynálezů zaručující vlastníkovi patentu výhradní právo k průmyslovému využití vynálezu. V České republice udělování patentů upravuje zákon 527/1990. Podle něj se patenty udělují na vynálezy, které jsou nové, jsou výsledkem vynálezecké činnosti a jsou průmyslově využitelné. Vynález se považuje za nový, jestliže není součástí stavu techniky. Stavem techniky je všechno, co bylo zveřejněno přede dnem přihlášení patentu, ať již v České republice nebo v zahraničí. 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Maximální možná délka patentové ochrany je 20 roků. http://cs.wikipedia.org/ United States Patent and Trademark Office www.uspto.gov European patent office www.epoline.org Úřad průmyslového vlastnictví www.upv.cz www.ctt.muni.cz Brno, No. 27 on the list of 52 places to visit in 2016 www.ce-ce.org CECE 2018 15th International Interdisciplinary Meeting on Bioanalysis Brno, October 15 - 17, 2018 Brno, among Top 10 Student Cities According to Students in 2018 CECE 2020 Siem Reap, CambodiaCECE 2020 Siem Reap, Cambodia ITP/CECE 2021 Brno, Czech RepublicITP/CECE 2021 Brno, Czech Republic