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. U.S. patent grants are effective only within the United States, U.S.
territories, and U.S. possessions. Under certain circumstances, patent term extensions or
adjustments may be available.
What is granted is not the right to make, use, offer for sale, sell or import, but the right to exclude
others from making, using, offering for sale, selling or importing the invention. Once a patent is
issued, the patentee must enforce the patent without aid of the USPTO.
There are three types of patents:
1) Utility patents may be granted to anyone who invents or discovers any new and useful
process, machine, article of manufacture, or composition of matter, or any
new and useful improvement thereof;
2) Design patents may be granted to anyone who invents a new, original, and ornamental
design for an article of manufacture; and
3) Plant patents may be granted to anyone who invents or discovers and
asexually reproduces any distinct and new variety of plant.
Patentable subject
1. 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.
Za vynálezy se naopak nepovažují zejména :
objevy, vědecké teorie a matematické metody,
pouhé vnější úpravy výrobků,
plány, pravidla a způsoby vykonávání duševní činnosti,
programy počítačů,
pouhé uvedení informace
Majitel patentu má výlučné právo vynález využívat (tj. výrobek vyrábět, uvádět do oběhu nebo
upotřebit postup), dále poskytnout souhlas k využívání vynálezu jiným osobám (např. licenční
smlouvou) a má právo převést patent na jinou osobu.
Proto, aby patent zůstal v platnosti, je nutno platit tzv. udržovací poplatky, a to v každém státu
zvlášť. 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
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