Tuesday, August 5, 2014

Princeton University – Plundering of Science I


Pursuing a balance with NATURE

_____________________________________________________________

Hydrology Breakthrough vs. USPTO Conspiracy


"Violation of science: bad apples and/or systems failure?"

Why Bad Apples Spoil the Barrel?


The Conspiracy – USPTO is running a broad Reinvention Scheme that let Lawyers writing scientific patents and allowing IP rights of issues they are not known in the art!


‘...  If you have a point to make about my treatment of hydrological concepts, I ask that you take the time to explain your specific points of disagreement.  I note that my work is better represented in my publications (available at http://www.stroockgroup.org/home/publicationsthan in patents, as the lawyers have been translated the latter into legalese that I do not understand.

Food for thought – If an inventor PhD from Harvard can have USPTO issued patents to protect his intellectual property rights but he does NOT understand it, how much the Patent Attorney and Patent Examiner knows about the issue granted protection by US Government. If it holds true, it means that Lawyers know about science more than scientists doAmazing! I just imagine Albert Einstein trying to get patents with his scientific papers .. . It means that those guys in the patenting affairs would handle Theory of Relativity more than him . . . Now I understand why Americans are the leaders of the world . . . awesome, this is simply magical. Mayday Mr. Snowden .  . .mayday mother nature!

My ‘scientific breakthrough’ deals deeply with Hydrogeology/Soil Physics/Hydrology. When I took classes at the Pennsylvania State University of such disciplines during my PhD in Soil Science I remember seeing no single Law student as classmate.

If my patent was being violated I had simple questions to pursue:

·       Was it a casual violation or a clear biased trend?
·       Had the examiner already cited my patent earlier?
·       Did the inventors and examiner have technical-scientific background in the issue?
·       Was the violator a wealthy party?
·       Was the examiner citing my patent to be sure he was granting new claims not claimed before?
·   Why my issued patent was being randomly cited for irrelevant patents and ignored when violated blatantly? 
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This is the tip of the iceberg as of what lies  bellow, clearly points out distorted conceptions and beliefs on human culture WHEN LAWS OF MAN IGNORE LAWS OF NATURE.


From: Moulis, Thomas <
Thomas.Moulis@uspto.gov>
 
Date: Fri, Aug 7, 2015 at 12:40 PM 
Subject: RE: Conspiracy and Brainwashing III – USPTO is preaching to their Patent Examiners that they do not need to be known in the art for judging and allowing IP rights when issuing patents! 
To: Elson Silva, PhD <tubarc@gmail.com>

You are a fool--- 

If you can’t understand legal or technical writing, you have no business blogging about it 

Wicking” is a term of art---fluid will travel in any direction via the fibers—regardless of gravity

-------------------

"Wick/wicking is in the patent classification system but not on HYDROLOGY textbooks."


LEGAL WICKING as the term of the art regardless of gravity confirms USPTO long standing bias ignoring hydrology on conductivity parameters of issued patents (11/30/23):


Thermal/Heat Conductivity                       176.074 pat.

Electric/Electrical Conductivity                139.254 pat.

Hydraulic Conductivity                                  1.329 pat.

Unsaturated Hydraulic Conductivity                  38 pat.

Wick/wicking                                                      66.415 pat.


The US Government states that LEGAL WICKING is not technical, being inert to gravity LAW, meaning that LEGAL OIL LAMPS and LEGAL CANDLES can work upsided down. This sort of deceiving is behind the Economic Melting Down of 2008 burning about 41 trillion dollars, also dumping 1,1 million American as the leader of the COVID-19 pandemic catastrophe that  took around 7 million lives world widely. In addition, obesity and sedentarism letting human beings miss brain capacity by becoming grumpier and dumber on neurogenesis effect.

Science is our understanding on nature functioning. Humans learn to respect nature early as babies on the first steps taming gravity for walking and running. Soon we understand the consequences of missteping and falling down. Therefore, all issued patents dealing with wick/wicking are CERTAINLY frauded because PATENT EXAMINERS ignored their homework from the beginning of their lives - GRAVITY

Sir Isaac Newton defined the Law of Universal Gravitation in 1687. He was inspired to formulate his theory of gravitation by watching the fall of an apple from a tree.

---------------------------------
De: Owen, Steven [mailto:steven.owen@uconn.edu]
Enviada em: quarta-feira, 5 de outubro de 2011 11:36
Para: Elson Silva, PhD
Assunto: RE: {SPAM?} Protecting Hydrology Science from REINVENTION

Mr. Silva, has anyone ever called you a nutcase

Are people out to get you? 

Are you having some trouble keeping up with your medications?


---------------------------------
Steven V. Owen
University Professor Emeritus
Educational Psychology

---------------------------------

Dr. Owen, my medicine is a bit bitter than that one swallowed by Albert Einstein by just stretching his tongue:

http://youtu.be/E3d-JRg28p8

https://youtu.be/czv2OiiC5wA


_____________________________________________________________


This email from an Emeritus Faculty of Educational Psychology provided valuable insights and feedback showing how deep the academic community is compromised on scientific affairs in the US.They were supposed to know that Darcy’s Law on Hydraulic Conductivity is not written in the US constitution, but endorsed by Mother Nature.


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Nobel Prize is IMPORTANT TO HUMAN KIND, as 50% is business, 40% politics, 5% bad science and 5% good science from an educated abstraction. Nobel Prize Nomination of Economic Science was introduced to pretend that Human Business could overlap nature functioning. THE US AS THE FIRST ECONOMIC POWER IN THE WORLD GRABBED TWO-THIRDS (411/621) OF NOBEL PRIZE NOMINATIONS FOR SCIENCE. Exploratory Analysis shows that the American scientific community has been violating Hydrology science in the Patenting System more than a century, leaving a gap huge enough for a new science Hydrotechnology.  It seems that working with the Chemistry of explosives is far more profitable than the Hydrology of self-watering flower pots. Mr. Alfred Nobel, Arms Dealer, Merchant of Death, and Father of Dynamite, got 355 patents and Albert Einstein got 50 issued patents to portray top scientists claiming intellectual property rights. Obesity, Economic Melt Dow, COVID-19 tragedies, and now Reinvention Policy by USPTO are important evidence of American negligence to science misbalance with Nature.

How much NATURE endorses the Economy and Politics? Sunlight and rain come to us FREE OF CHARGE, regardless of BOUNDARIES, making the Economy not a science, but a distorted human affair as basic Laws of offer and demand is being replaced by GREED and FEAR. Likewise,  recent wars in Ukraine and Israel show us that Politics can’t be science, but a wicked manipulation on human issues wasting innocent lives and spoiling the landscape for weaponry industry profit and disguised interest as Homo sapiens misses simple rationality.

In my neighborhood, I saw the Scientific Police taking pictures of swings I installed on trees for children in the Park during the COVID-19 pandemic lock down. Society try to employ the word SCIENCE for POWER, but there is a misunderstanding as scientific principles claim TRANSPARENCY and HONESTY. Nature is in charge of SCIENCE as there is no POLICE to enforce Nature LAWS. Even religion try to use Scientology for credibility. I like the simple conception that God = Nature. However, Nature writes no books, promises no lands, no life after death, no war or death in name of a divine. In around 4 billions of years of our planet, it seems that we got no aliens to affect our evolution. Most probable we are not leaving our home until the end in 4 to 6 billions of years. Human challenge is to keep nuclear weapons safe, cropping soils, mining our minerals and preserve our home in balance with nature functioning, making our blue planet good for all humans. 

What we see in the universe is just for light travelling.

It seems that few scientists do understand the meaning of their titles PhD as Philosophy Doctor coming from Philosophy of Science (Epistemology, Metaphysics, Logics, and History of Science).

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The Conspiracy
_________________________________________________________________

Princeton University  Plundering of Science

Is it possible to reinvent a ‘scientific breakthrough’ in Hydrology if you are a Physicist having no expertise in the issue?

Who cares?

I got my PhD at a famous place Penn State that let boys be raped in the locker rooms more than a decade. Bill Clinton then President was on my commencement saying we would be inventing new technologies never imagined before.  He did not mention about fixing government malfunctioning violating science in the core! While money was poring and fame was high Joe Pa and Graham Spanier did not budge to protect a sound functioning of society as it seems to be a major trend around . . . after all who cares about disadvantaged boys and even less for science?

USPTO Reinvention Policy

Inventors owning US patents only collect money if they can afford court fees suing violators reinventing their ideas over and over sometimes by flawed patents from lay inventors and wealthy parties.

What does it means?
It means that inventions granted American patents might collect rewards if the inventor can afford 2 to 5 million dollars for suing a continuous reckless patent infringement from wealthy parties colluding with US Officials.

American patents are becoming useless if the inventor cannot afford steep court fees on continuous litigations.

Who takes advantages by the Reinvention Policy?
US government and Law Firms profit high by reinvention policy spoiling the patenting system and harming inventors that cannot feed such swindle.

Who are the Patent Examiners and what is their technical-scientific background?
USPTO confirmed having no single Patent Examiner with background in Hydrology among around 6,000 examiners. So, Patent Examiners are simply not known in the art for judging the issue granted!

I told Mr. Obama early 2011 not to dishonor Hydrology this way . . .  http://hydrotechnology.blogspot.com.br/


Corporation Pillage on Inventions

EcolabFlawed Violations by wealthy corporations
Ecolab employs more than 26,000 people with more than $6 billion in global sales. I sent letters to Ecolab, to Kinney & Lange, P.A the patent law firm, and to USPTO about violating my patent rights with a flawed one that even does not work.
The fluid only drops after crossing the water table as portrayed in the arrow reference because the ‘wick’ needs to cross the boundaries between Unsaturated and Saturated Hydraulic Zones for water to drop. The aggravation is that after I notified all parties in written about the violation of IP rights and scientific flaws more patents applications were filed and issued US 7,285,255 and US 7,670,551 . This is just unacceptable. What kind of protection anybody can get by ignoring IP rights and scientific literature?



video can show it in 5 minutes what lay people missed in a patenting approval.

Hydrology Science is being shamefully ignored in the Patenting System with flawed patents that violate common knowledge from scientific literature and issued patents. It seems that sometimes the Patent Examiner (US pat. 7,285,255) violating science and issued patents is a director meaning that lay people handling Hydrology at USPTO is a critical bias going up in the chain of power by people that are overstepping Ethical grounds beyond their due expertise. This was not a simple mistake by a junior Patent Examiner since Ms. Gladys Corcoran had issued 202 patents so far.

REINVENTION POLICY BY USPTO: When a Patent Supervisor and Director (Ms Gladys Corcoran) issues a flawed patent US 7,285,255 hurting Hydrology and my issued patents it points the tip of the iceberg portraying a wicked reinvention policy by USPTO. 
Ms Gladys J. P. Corcoran and Mr. Sean E Conley Primary and Assistant Examiner four months earlier issued 7,244,398 (July 17, 2007) citing my issued patent 6,766,817 meaning that they were aware of the new conceptions in hydrodynamics disclaimed in the art if they were knowledgeable in the art of Hydrology. Consequently the flaw issued at US 7,285,255 (October 23, 2007) was not a simple consequence of careless professionals working off their expertise boundaries ignoring advanced Hydrology already on literature and updated by 6,766,817very likely un underlying consistent bias breaking the Law on Ethic boundaries.. 
Mr. Gregory Morse a Patent Commissioner answered my letter saying that there is a Law stating that the drawing dimensions does not need to represent the real dimensions of the device. I replied saying that I learned in High School about scale that all details are represented in the same proportional ratio and that the scientific flaw of (US pat. 7,285,255) takes places at any scale like parallel lines that never meet each other at any scale employed.
I got PhD in Soil Science/Spatial Applied Hydrology with a deep background in Remote Sensing and GIS and now I have my ‘scientific breakthrough’ being violated by lazy patents from people that their background is lower than High School standards.

‘It seems that Mr. Clinton wasted his time on my commencement.’

Roche employs about 67,000 people around the world and has about 44 billion dollars revenue annually. A group of researchers at Roche Diagnostics was filing dozens of patent applications trying to develop the spatial geometry of a channel on lancets to conduct blood for glucose test.

This left figure shows that Roche does not employ any Hydrologist or Soil Scientist to understand the deep functioning of fluids moving on porous systems, even less the rounding geometry. So, they learned with my patent, copied it and poorly reinvented it certainly bribing and corrupting US officials, as well as ignoring my pleas for HONESTY under the Law. The violation is so despicable that the Patent Examiner Mr. Max Hindenburg allowing the violation had already cited my patent earlier allowing 7,377,904 on May 27, 2008 and the technical background is so poorly handled that the Hydrology scientific community is going to become mesmerized by such nuisance

I told Roche and put more than 4,000 of their emails on my blog (http://roche-tubarc.blogspot.com ) that If they have this power to bribe US officials on money making, they should be smarter and bribe people at the Treasure Office by printing the bills. Stealing my invention poorly is just impossible as knowledge cannot be recreated even less by those ones not known in the art missing critical background and expertise. 


Carefusion Corporation employs around 16,650 people and has an annual budget of about 4 billion dollars revenue. It filed seven patents poorly claiming the same geometry of Tubarc.

US 20130255672 - TRANSPORTING LIQUID IN A RESPIRATORY COMPONENT 
Abstract
A respiratory component coupled with a breathing circuit, the respiratory component including at least one groove disposed upon a surface of the respiratory component, wherein the respiratory component is not a heater wire.
Claims
1. A respiratory component comprising: a liquid transporting device configured for transporting a liquid from a first region to a second region, wherein said liquid transporting device is disposed on a surface of said respiratory component and is not disposed on a heater wire.

2. The respiratory component of claim 1, wherein said liquid transporting device comprises: at least one groove disposed upon said surface of said respiratory component, wherein said at least one groove is configured for wicking up liquid from said first region and transporting said liquid to said second region. 

Illumina, Inc.They got the same geometry as Tubarc twice. As a scientist I am glad to have created something that is really good and important to human kind, but I was expecting a reward from it so I could invest in my research project. It means that the Intellectual Property system is failing to scientists and inventors on their dedication and hardship.



Abstract
A fluidic device for conveying liquid to a well of a microplate. The device includes a support structure configured to be mounted along the microplate. The device also includes a microfluidic tube coupled to the support structure. The tube has an inlet, an outlet, and an open-sided channel that extends longitudinally therebetween. The tube has a cross-section that includes an interior contour with a gap therein. The gap extends at least partially along a length of the tube. The tube is configured to convey liquid to the well of the microplate when the tube is held in a dispensing orientation.

Claims
1. A fluidic device for conveying liquid to a well of a microplate, the device comprising: a support structure configured to be mounted along the microplate; and a microfluidic tube coupled to the support structure, the tube having an inlet, an outlet, and an open-sided channel that extends longitudinally therebetween, the tube having a cross-section that includes an interior contour with a gap therein, the gap extending at least partially along a length of the tube, the tube being configured to convey liquid to the well of the microplate when the tube is held in a dispensing orientation. 


Academic Pillage on Hydrology Science



UCONN IIIIIIIVVVIVIIVIIIIXXXIXIIXIIIXIV

Academic Dishonoring of a scientific breakthrough

Dr. Amir Faghri and Dr. Zhen Guo working at the former Fuel Cell Center at the University of Connecticut believed that Hydrology from their Library can be ignored as well as my patent rights at USPTO when issuing US 7,625,649 ‘Vapor feed fuel cells with a passive thermal-fluids management system’ below. It was sent many letters requesting IDS of US 6,766,817 to many faculties and Intellectual Property management team at Univ. of Connecticut. Nobody there felt any urge to respect neither Classical Hydrology nor already issued patents on the subject. There is no doubt about breach of Ethics and breaking the Law by reinventing something already invented.

It seems that Mechanical Engineers insist on working with fluids having a private Hydrology on their own ignoring Classical Hydrology from old textbooks. US 6,766,817 not only designed the interplay between Hydraulic Zones allowing self-sustaining reversible flow, but also it gauged Unsaturated Hydraulic Conductivity as 2.18 mm/s, besides Tubarc as an enhance microporosity geometry for fluid conduction.

Looking at the drawings I have a deep feeling that it was a straight lousy copycat by a PhD faculty showing arrogance and how stupid he can be by ignoring a ‘scientific breakthrough’.

Is US 6,766,817 a ‘scientific discovery’?
I believe so because so far only 24 issued patents had mentioned Unsaturated Hydraulic Conductivity while wick/wicking from lay people was mentioned in 26.852 issued patents on Mar 24, 2011.


Collapsing Scientists – Renowned scientists are developing know-how on fluidic devices ignoring basic understanding on Hydrology principles and IP rights on issued patents by USPTO. The US pat. Appl.  20140191438 - Microfluidic Devices and Methods of Fabricationhas neither a single quotation on ‘hydrology’ nor any Hydrology reference complying with the main science of fluids.
hy•drol•o•gy - The scientific study of the properties, distribution, and effects of water on the earth's surface, in the soil and underlying rocks, and in the atmosphere.

How can any scientist address fluidic devices without knowing Hydraulic Conductivity (K) on Hydrodynamics? Hydraulic Zones? Hydraulic Gradient?

Hydraulic Conductivity (K) = volume / area / time 

Scientific Flaw


They are not aware of employing ‘Unsaturated Hydraulic Siphon”  to displace fluids by a hydraulic gradient on the interplay between Hydraulic Zone when fluids can move reversibly by unsaturated flow taking advantage of molecular connectivity of fluid dynamics.




SUNNY IIIIIIIV

Dr. Gorfinkel Mathematician/Physicist from SUNY (US 8,231,844) reinvented Tubarc (US 6,766,817having no single paragraph addressing Hydrology Science on her issued patent. I am curious on how much Hydrology is taught in Mathematics and Physics courses to give professionals required background to handle such issue.

Title: Method and device for manipulating liquids in microfluidic systems. Assignee: The Research Foundation Of State University Of New York (AlbanyNY)



Complex Engineering on hydrodynamics is confused on geometries between squares and circles!

Dr. Fedder, PhD, is proposing a squared geometry in order to control fluid dynamics employing ‘Capillary Action’which comes from the cylindrical structure conception – capillary’
How much brain power is required to realize that ‘capillary action’ is a cylindrical geometric conception to capture unsaturated hydraulic flow when the solid surface of porosity pulls fluids upward against gravity having a suction or negative hydraulic flow? Frankly speaking can any capillary be squared? It seems that Nature opted for rounded geometries since the beginning making the sun, earth, cells, veins, trunks, etc, all rounded and rounded. Dr. Fedder empty skull is also rounded!
BTW – Dr. Fedder did not allow his patent application be published and it was issued seven years afterwards!
Carnegie Mellow is just raping science and shamefully cheating on Academic Affairs in collusion with USPTO shameful reinvention policy. Dr. Gary K. Fedder the Director of the Institute for Complex Engineered Systems (ICES) got B.S. and M.S. from MIT in 1982 and 1984,obtaining in 1994 PhD at the University of California at Berkeley.

UC SANTA BARBARA IIIIII


US Pat. Application 20130327504 - TITANIUM-BASED THERMAL GROUND PLANE assigned to THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, Oakland, CA - December 12, 2013.
The word wick is not portrayed in any Hydrology textbooks because fluid moves in response to a Hydraulic gradient far more complex than oil lamps and candles requiring no flames at all. The patent application from University of California mentions the word wick 128 times while the word Hydrology is ignored shamefully by scientists pretending to know what they are doing.


Dr. Austin is a Professor of Physics trying to reinvent tubarc (US 6,766,817) ignoring Hydrology shamefully.
If for a lay person in Hydrology like a Physicist or a Mathematician channels or trenches are just containment structures, but for a Hydrogeologist it is a conceptional continuous pore connectivity to favor higher level of anisotropic unsaturated hydraulic flow providing higher reliability and enhanced control of hydrodynamic properties for molecular connectivity on porosity systems. Capillarity based on tube theory does not allow a making of a porosity for having one directional flow inside the tube. Tubarc (tube+arc) allows multidirectional flow with prevailing anisotropic longitudinal flow. It looks like simple and obvious, but Nature designed biological porosity with channels and perforations to be sure that fluids flow moving substances around as needed.
Tubarc = alignment of pores longitudinally bearing high geometrical pattern for fluid conduction harnessing Unsaturated Hydraulic Flow and producing an enhanced porosity structure with high control on void ratio and reversible fluid displacement in the interplay between Hydraulic Zones.
Sometimes we must refrain from saying how stupid somebody is simply for not bearing enough brain power from such comprehension. Nature endows brain to all human beings, but their deep functioning varies a lot. Perhaps intelligence goes far beyond IQ levels and can be perceived on expanding waistlines to portrait an understanding if brain resources are applied to self-maintenance on a modern different landscape we are molding day by day that challenge our deep physiology even more from the past we came from.

They are violating my IP rights simply for not having enough brain power to comprehend the principles and even less about honesty molding values for respect and dignity. Indeed intelligent people do not do such a thing, as they know deeply about coherence, harmony, logics, and values! I believe that a smart man would never dare to cheat Albert Einstein achievements.
Robert H. Austin received his B.A. in Physics from Hope College in Holland MI and his Ph.D. in Physics from the University of Illinois Champaign-Urbana in 1976. He was a post-doc at the Max Planck Institute for Biophysical Chemistry from 1976-1979 and has been at Princeton University in the Department from Physics from 1979 to the present, achieving the rank of Professor of Physics in 1989.

Position: Professor
Title: Professor of Physics.
Office: 122 Jadwin Hall 
Phone: 609-258-4353 


US Pat Appl 20140206555 - Nanochannel Arrays and Their Preparation and Use for High Throughput Macromolecular Analysis 
Abstract
Nanochannel arrays that enable high-throughput macromolecular analysis are disclosed. Also disclosed are methods of preparing nanochannel arrays and nanofluidic chips. Methods of analyzing macromolecules, such as entire strands of genomic DNA, are also disclosed, as well as systems for carrying out these methods.
Claims



1. A method of analyzing at least one macromolecule, comprising the steps of: transporting at least one macromolecule in a fluid sample into a nanochannel, such that the at least one macromolecule is in an elongated state within the nanochannel; maintaining the at least one macromolecule in an elongated state within a region of the nanochannel having a substantially constant width; detecting at least one signal from the at least one elongated macromolecule within the nanochannel; and correlating the detected signal to at least one property of the at least one macromolecule.



2. The method of claim 1, wherein the at least one macromolecule is transported into the nanochannel by applying an electric current.



3. The method of claim 1, wherein the trench depth of the nanochannel is less than about 200 nanometers.



4. The method of claim 1, wherein the step of maintaining the at least one macromolecule in an elongated state within the nanochannel is achieved via confinement of the at least one macromolecule within the nanochannel.



5. The method of claim 1, wherein the nanochannel comprises dimensions that are at most the same order of the persistence length of the macromolecule.



6. The method of claim 1, wherein the step of transporting the at least one macromolecule into the nanochannel such that the at least one macromolecule is in an elongated state occurs in a first region of the nanochannel and the step of detecting the at least one signal is performed in a second region of the nanochannel.



7. The method of claim 1, wherein the step of maintaining the at least one macromolecule in an elongated state within the nanochannel is achieved without the requirement of shear force acting on at least one macromolecule.


8. The method of claim 1, wherein the nanochannel is in the material of a surface.

9. The method of claim 1, wherein the nanochannel is surmounted by sealing material to render the nanochannel at least substantially enclosed.


16. The method of claim 1, further comprising the step of illuminating the at least one macromolecule within the nanochannel.

17. The method of claim 10, wherein the sealing material is optically transparent, and wherein the signal is transmitted through the optically transparent sealing material.

18. A method of maintaining at least one macromolecule in an elongated state, comprising the steps of: transporting at least one macromolecule in a fluid sample into a nanochannel, such that the at least one macromolecule is in an elongated state within the nanochannel; and maintaining the at least one macromolecule in an elongated state within a region of the nanochannel having a substantially constant width.

19. The method of claim 18, wherein the at least one macromolecule is transported into the nanochannel by applying an electric current.

20. The method of claim 18, wherein the trench depth of the nanochannel is less than about 200 nanometers.

21. The method of claim 18, wherein the step of maintaining the at least one macromolecule in an elongated state within the nanochannel is achieved via confinement of the at least one macromolecule within the nanochannel.

22. The method of claim 18, wherein the nanochannel comprises dimensions that are at most the same order of the persistence length of the macromolecule.

23. The method of claim 18, wherein the step of maintaining the at least one macromolecule in an elongated state within the nanochannel is achieved without the requirement of shear force acting on at least one macromolecule.

24. The method of claim 18, wherein the nanochannel is in the material of a surface.

25. The method of claim 18, wherein the nanochannel is surmounted by sealing material to render the nanochannel at least substantially enclosed.

 [0002] The present invention relates to a nanochannel array. The present invention also relates to a method of preparing nanochannel arrays. The present invention also relates to nanofludic chips containing nanochannel arrays. The present invention also relates to a system suitable for high throughout analysis of macromolecules. The present invention also relates to a method of analyzing at least one macromolecule by using a nanochannel array.

[0003] In the newly emerging field of bioanotechnology, extremely small nanofludic structures, such as channels, need to be fabricated and used as arrays for the manipulation and analysis of biomolecules such as DNA and proteins at single molecule resolution. In principle, the size of the cross sectional area of channels should be on the order of the cross sectional area of elongated biomolecules, i.e., on the order of 1 to 100 square nanometers, to provide elongated (e.g., linear, unfolded) biomolecules that can be individually isolated, yet analyzed simultaneously by the hundreds, thousands, or even millions. Likewise, it is also desirable that the length of the channels should be long enough to accommodate the longest of macro-molecules, such as an entire chromosome, which can be on the order of 10 centimeters long (e.g., chromosome 1 of the human genome having 250 million base pairs). The present inventors and others have recently been concerned about such problems and their possible solutions, as reported in: O. Bakajin, et al, Anal Chem, 73 (24), 6053 (2001), J. O. Tegenfeldt, et al, Phys. Rev. Lett. 86 (7), 1378 (2001), J. Han et al., Science 288, 1026 (2000), and S. R. Quake et al., Science 290 (5496), 1S36 (2000).

[0004] It is important to efficiently and reliably construct arrays of many thousands, or even millions of channels in an array for the simultaneous isolation and analysis of up to thousands or millions of individual macromolecules. Such large arrays of isolated macromolecules could, in principle, be analyzed with presently available two-dimensional area detectors, such as charge-coupled devices (CCDs). Together with automated data-processing collection and image analysts software, the simultaneous characterization of up to thousands or millions of micromolecules would be an extremely powerful tool for macromolecular analysis, such as population distribution analysis of macromolecular size, chemical composition, and DNA sequencing.

[0005] Because individual macromolecules could in principle be isolated and analyzed in a single channel, heterogeneity of a sample containing a multitude of macromolecules can be readily discerned. This would be particularly useful for identifying single nucleotide polymorphisms (SNP) on a single chromosome. In contrast, traditional population based assays require time-consuming DNA amplification methods to prepare multiple copies of a nucleic acid macromolecule to carry out SNP analysis. If available, a chromosomal analysis system incorporating nanochannel arrays could perform SNP analysis much more quickly than any method presently available.

[0006] Nanochannel arrays having the proper dimensions for carrying out the simultaneous isolation and analysis of a multitude of elongated macromolecules have been heretofore unavailable. Accordingly, there is an urgent need to provide nanochannel arrays having at least three key dimensional qualities: (1) the channels should have a sufficiently small dimension to elongate and isolate macromolecules; (2) the channels should have a sufficiently long dimension to permit the instantaneous observation of the entire elongated macromolecule; and (3) a high number of channels should be provided to permit the simultaneous observation of a high number of macromolecules. In addition, it would be desirable for the elongated and isolated macromolecules to remain indefinitely in such a state at ambient conditions even after the field which is used to transport the macromolecules into the channels (e.g., electric field) is turned-off. This feature would permit the macromolecules to be analyzed with techniques that require times longer than the residence time of the macromolecules under the influence of the field. This feature would also permit the analysis of macromolecules without having to subject them to a field.

[0007] Methods for analyzing macromolecules (e.g., polymers) have been previously disclosed, however none uses a nanochannel array having the three key dimensional qualities as described supra. U.S. Pat. No. 5,867,266 discloses a micro optical system having a plurality of coplanar micron-to-millimeter scale sample channels prepared using photolithography and an artificial gel material comprising a multiplicity of pillar structures in each micron-to-millimeter wide sample channel. The large channel width makes this system unsuitable as a nanochannel array.

[0008] Likewise, methods for analyzing macromolecules (e.g., polymers) by isolating them in channels more narrow than this disclosed in U.S. Pat. No. 5,167,266, however none uses a nanochannel array having the three key dimensional qualities as described supra. In WO 00/09757, several of the inventors of the present patent application disclose a system for optically characterizing single polymers that are transported in a straightened form through a channel. In U.S. Pat. No. 6,355,420, a system is disclosed for analyzing polymers that are transported in a straightened form through a plurality of (at least 50) channels. While both of these disclosures are directed towards analysis of single macro-molecules aligned in one or more channels, neither of these documents discloses the simultaneous observation of a high number of macromolecules in a multitude of channels.

[0009] Thus, there remains the problem of providing suitable nanochannel arrays that are useful in a variety of macromolecular analysis. Methods for analyzing macromolecules (e.g., polymers) by isolating them in a narrow channel have been previously disclosed, however none uses a nanochannel array having the three key dimensional qualities as described supra, primarily because, until now, fabrication techniques for constructing such a nanochannel array were not available.

[0010] In creating ultra-small nanofluidic structures, e.g., for single biomolecule analysis, at least two problems need to be solved: reduction of size and creation of sealed fludic channels. As reported by one of the present inventors, NIL is a parallel high-throughput technique that makes it possible to create nanometer-scale-features over large substrate surface areas at low cost. (S. Y. Chou et al. Appl. Phys. Lett 67 (21), 3114 (1995) and S. Y. Chou et al. Science 272, 85 (1996)) Current sealing techniques such as wafer bonding (M. Stjernstrom et al., J. Micromech. and Microeng. 8 (1), 33 (1998)), and soft elastomer sealing (H. P. Chou et al. Proc. Nat. Acad. Sci. USA 96 (1), 11 (1999), are suitable for relatively large planar surfaces and provide an effective seal. Wafer bonding requires an absolutely defect tree and flat surface, and elastomer sealing suffers from clogging due to soft material intrusion into the channels. Within extremely small confining structures biological samples are also much more sensitive to issues such, as hydrophobicity and the homogeneity of the material constructing the fluidic structure.

[0011] Recently developed techniques using "place-holding" sacrificial materials such as polysilicon (S. W. Turner et al. J. Vac. Sci, and Technol. B. 16(6), 3835 (1998)) and polynorbornene (D. Bhusari et al., J. Microelectromech. Syst. 10 (3), 400 (2001)) have gained popularity to create sealed small, hollow fludic structures. However, steps needed in removing the sacrificial materials such as heating the substrate up to 200-400.times. or wet etching limits the use of certain materials and downstream fabrication processes.

[0012] As provided herein, the present invention achieves the goal of providing nanochannel arrays suitable for performing high throughput macromolecular analysis. Interferometric lithography (IL), nanoimprint lithography (NIL), and non-isotropic deposition techniques are used to prepare nanochannel arrays having hundreds of thousands to more than a million enclosed channels having the desired key dimensions across the surface of a silicon wafer substrate.

[0013] In one aspect of the present invention, there are provided nanochannel arrays including a surface having a plurality of channels in the material of the surface, said channels having a trench width of less than about 150 nanometers and a trench depth of less than 200 nanometers; at least some of the channels being surmounted by sealing material to render such channels at least substantially enclosed.

[0014] In a further aspect of the present invention, methods of preparing nanochannel arrays are disclosed, which include the steps of: providing a substrate having a surface; forming a plurality of channels in the material of the surface; and depositing a sealing material on the plurality of channels to surmount the plurality of channels to render such channels at least substantially enclosed, the substantially enclosed channels having a trench width of less than 150 nanometers and a trench depth of less than 200 nanometers.

[0015] In another aspect of the invention, there are provided nanofluidic chips including; a) nanochannel array, including: a substrate having a surface; a plurality of parallel channels in the material of the surface, said channels having a trench width of less than about 150 nanometers and a trench depth of less than 200 nanometers; at least some of the channels being surmounted by sealing material to render such channels at least substantially enclosed; at least some of the channels are capable of admitting a fluid; b) at least one sample reservoir is fluid communication with at least one of the channels, said sample reservoir capable of releasing a fluid; and c) at least one waste reservoir in fluid communication with at least one of the channels, said waste reservoir capable of receiving a fluid.

[0016] In yet another embodiment of this invention, there are provided systems for carrying out analysis. In exemplary embodiments, these include: A) a nanofludic chip, including: a) nanochannel array, including: a substrate having a surface, a plurality of parallel channels in the material of the surface, said channels having a trench width of less than about 150 nanometers and a trench depth of less than 200 nanometers; at least one of the channels being surmounted by sealing material to render such channels at least substantially enclosed; at least one of the channels capable of admitting a fluid; and b) at least one sample reservoir in fluid communication with at least one of the channels, said sample reservoir capable of releasing a fluid; and B) a data processor.

[0017] In another embodiment, methods of analyzing at least one macromolecule are described which, for example, include the steps of: providing a nanofluidic chip, including: a) nanochannel array, including; a surface having a plurality of parallel channels in the material of the surface, said channels having a trench width of less than about 150 nanometers and a trench depth of less than 200 nanometers; at least one of the channels being surmounted by sealing material to render such channels at least substantially enclosed; at least one of the channels capable of admitting a fluid; b) at least one sample reservoir in fluid communication, with at least one of the channels, said sample reservoir capable of releasing a fluid containing at least one macromolecule; providing the at least one sample reservoir with at least one fluid, said fluid comprising at least one macromolecule; transporting the at least one macromolecule into the at least one channel to elongate said at least one macromolecule; detecting at least one signal transmitted from the at least one elongated macromolecule; and correlating the detected signal to at least one property of the at least one macromolecule.

[0018] Cartridges including a nanofluidic chip in accordance with this invention are also disclosed herein. Such cartridges are capable of being inserted into, used with and removed from a system such as those shown herein. Cartridges useful with analytical systems other than the systems of the present invention are also comprehended by this invention.


[0019] FIG. 1 illustrates a cross-section of a nanochannel array having substantially enclosed channels.

[0020] FIG. 2 illustrates a cross-section of a nanochannel array having completely enclosed channels and having sealing material deposited in the channels.

[0021] FIG. 3 is a scanning electron micrograph of a nanochannel array having parallel linear channels and open channel ends.

[0022] FIG. 4 illustrates a schematic of a process for depositing sealing material into the channels.

[0023] FIG. 5 illustrates a nanofludic chip.

[0024] FIG. 6a is a scanning electron micrograph of the substrate used in Example 4 prior to sealing with silicon dioxide.

[0025] FIG. 6b is a scanning electron micrograph of the nanochannel array of Example 4 obtained after sealing the substrate with silicon dioxide.

[0026] FIG. 6c is a scanning electron micrograph (top view) of the nanochannel array of Example 5.

[0027] FIG. 6d is a scanning electron micrograph (top view) of the nanochannel array of Example 4.

[0028] FIG. 7a is a scanning electron micrograph of the substrate used in Example 1 prior to sealing with silicon dioxide.

[0029] FIG. 7b is a scanning electron micrograph of the nanochannel array of Example 1 obtained after sealing the nanochannel array in 7a with silicon dioxide.

[0030] FIG. 7c is a scanning electron micrograph of the substrate used in Example 2 prior to sealing with silicon dioxide.

[0031] FIG. 7d is a scanning electron micrograph of the nanochannel array of Example 2 obtained after sealing the nanochannel array of 7c with silicon dioxide.

[0032] FIG. 7e is a scanning electronmicrograph of the substrate used in Example 3 prior to sputtering with silicon dioxide.

[0033] FIG. 7f is a scanning electron micrograph of the nanochannel array of Example 3 obtained after sputtering the nanochannel array in 7e with silicon dioxide.

[0034] FIG. 8a illustrates a sealed channel having a nanoslit in an opaque layer across the bottom of a channel.

[0035] FIG. 8b illustrates a sealed channel having a nanoslit in an opaque layer across the sealing layer, which is oriented perpendicular to the long axis of a nanochannel.

[0036] FIG. 9a shows scanning electron micrographs of the substrate (left and bottom) and a of the sealed nanochannel array (right) used in Example 14.

[0037] FIG. 9b is the image obtained tram the CCD of the 48.5 kb lambda, phage genome (shorter) and 168 kb T4 phage genome (longer) of Example 14. Inset; Plot of genome size versus macromolecular contour length.

[0038] FIG. 9c shows a nanochannel array simultaneously elongating, separating, and displaying a plurality of DNA macromolecules ranging in size from 10 kb to 196 kb.

[0039] FIG. 10 illustrates a system for analyzing macromolecules using a nanofluidic chip.

[0040] One aspect of the present invention encompasses a nanochannel array having a plurality of channels that are substantially enclosed. As shown in FIG. 1, the nanochannel array 108 has a surface 102 that contains a plurality of channels 104 in the material of the surface 106. The channels 104 have a wall 110, and a channel center 112. The distance between the wall surfaces 110 inside a channel 104 that are perpendicularly opposite to the channel center 112 is defined as the trench width. The channels 104 are surmounted by a sealing material 188 that renders the channels 104 at least substantially enclosed.

[0041] In one embodiment, the channels 104 will not be completely enclosed and will typically have a sealing material 108 directly above the channel center 112, providing an opening in the sealing material to the channel. The opening may have a variety of shapes. The size of the opening is defined as the minimum distance spanning the opening above the channel center 112. In such embodiments, the size of the opening is less than the trench width, and is typically less than 1/2 of the trench width, more typically less than 1/3 of the trench width, and most typically less than 1/4 of the trench width, in other embodiments the channels can be completely enclosed, having sealing material completely covering the top of the channel and having no opening in the sealing material. In certain embodiments of the present invention, sealing material 108 can extend over the wails 110 and the bottom of the channels 104, as shown in FIG. 2. In such embodiments, the trench width is defined as the distance from the surfaces formed by the sealing material adjacent to the walls 114.

[0042] In the present invention, the trench width is typically less than 150 nanometers, more typically less than 100 nanometers, and even more typically less than: 75, 50, 25, and 15 nanometers. In certain embodiments the trench width is about 10 nanometers, in the present invention, the trench width is at least 2 nm, and typically at least 5 nm.

[0043] In the present invention the channels are at least substantially enclosed. "At least substantially enclosed" means that the channels are completely enclosed; or have an opening in the sealing material that is smaller than 1/2 the trench width, or have both completely enclosed channels and openings.

[0044] Channels that are completely enclosed have a trench depth that is defined as the distance between the surface of the solid material at the bottom of the channel below the channel center 112 to the sealing material above the channel center 112. Embodiments in which the channels having an opening have a trench depth defined as the distance from the surface of the solid material at the bottom of the channel below the channel center to the position of the opening where the opening size is measured. If the opening has more than one position where a minimum distance can be measured then the position of the opening is the one that is closest to the bottom of the channel 104.

[0045] In the present invention, the trench depth is less than 200 nm, in certain embodiments, the trench depth is typically less than 175 nm, and more typically less than 150 nm, 125 nm, 100 nm, 75 nm, 50 nm, and 25 nm. In certain embodiments the trench depth is about 1-5 nm. In certain embodiments the trench depth is at least 2 nm, typically at least 5 nm, and more typically at least 10 nm.

[0046] In the present invention, the nanochannel arrays can be formed in a substrate, such as a silicon wafer substrate, using a variety of fabrication methods as described below. In one embodiment, the nanochannel array has a plurality of parallel linear channels across the surface of substrate as illustrated by the scanning electron micrograph in FIG. 3.

[0047] In certain embodiments, the nanochannel arrays have at least one end of at least one of the channels can be in fluid communication with at least one reservoir. In these embodiments, at least one channel is connected directly with at least one reservoir. Alternatively, at least one channel can be connected with at least one reservoir via an interconnecting straight or curved microchannel (a channel having a width, height, or both larger than about a micron), or a channel is connected with at least one reservoir via an interconnecting nanopillar or micropillar array.

[0048] In certain embodiments, at least two ends of some of the channels are in fluid communication with at least one reservoir common to the channels. In these embodiments, at least two ends of some of the channels can be adjacent or not adjacent. These channels can be connected directly with at least one reservoir.

[0049] In certain embodiments, at least two channels can be connected with at least one reservoir via a common interconnecting straight or curved microchannel. Alternatively, at least two channels can be connected with at least one reservoir via a common interconnecting nanopillar or micropillar array.

[0050] In certain embodiments of the present invention, the nanochannel array has a plurality of channels that are in fluid communication with at least one sample reservoir common to at least some of the channels. By "a plurality of channels" is meant more than two channels, typically more than 5, and even typically more than 10, 100, 1000, 10,000 and 100,000 channels. In certain embodiments, one sample reservoir can be provided for all of the channels in the nanochannel array, thus the plurality of channels can be as large as the number of channels that are on the substrate, in a certain embodiments, 100 mm diameter substrates can have about 500,000 parallel linear channels having a periodicity of 200 nm, the periodicity being defined as the distance, between the middle of two adjacent channels.

[0051] In certain embodiments, the plurality of channels can be connected directly with at least one reservoir. The connections can be a common interconnecting straight or curved microchannel. In other embodiments, a plurality of channels can be connected with at least one reservoir via a common interconnecting nanopillar or micropillar array.

[0052] In certain embodiments of the present invention, the nanonanochannel array contains a plurality of channels that are in fluid communication with at least one waste reservoir. Although the plurality of channels is typically connected directly with at least one waste reservoir, more than one waste reservoir can also be provided. It should be appreciated that the waste reservoir can be used as a sample collection reservoir. Accordingly, multiple sample collection reservoirs can also be provided on nanochannel arrays. In these embodiments, a plurality of channels can be connected with at least one waste reservoir via a common interconnecting straight or carved microchannel as described earlier. Likewise, a plurality of channels can be connected with at least one waste reservoir via a common interconnecting nanopillar or micropillar array.

[0053] In certain embodiments of the present invention, the nanochannel array has a plurality of channels that are substantially parallel, in which the plurality of channels are substantially in the plane of the surface of the substrate.

[0054] In certain embodiments of the present invention, the nanochannel array can contain linear channels. Linear channels are adjacent channels that are substantially not interconnected.

[0055] In certain embodiments, the ends of the channels are capable of admitting a macromolecule in a fluid. By being capable of admitting macromolecule means that the channels have at least one opening large enough, to permit the passage of a macromolecule. While a variety of openings are envisages, typically such openings can be located at the ends of the channels or on the surface of the sealing material through openings in the sealing material. Openings in the sealing material can be provided by subsequent modification of the nanochannel arrays as provided below.

[0056] In certain embodiments of the present invention, the nanochannel array contains channels that are capable of transporting a macromolecule across their length. The nanochannel arrays can be fitted with a variety of components to affect macromolecular transport, examples of which include pressure or vacuum gradient drop across the channels, electroosmosis, and electrokinesis.

[0057] While not being bound to a particular theory, it is believed that macromolecules typically have a non-linear, three-dimensional conformation in space (e.g., linear polymers have a random coil conformation in their natural state). Accordingly, it is thermodynamically unfavorable for macromolecules to spontaneously elongate and enter channels directly from the environment due to the large free energy needed to reduce entropy. For example, a 169 kilobase T4 phage double stranded genomic DNA chains will form a Gaussian coil of radius of gyration Rg=(Lp/6).sup.1/2=700 nm in free solution, where L is its calculated contour length and p is the persistence length of about 50 nm.

[0058] In certain embodiments, the nanochannel array can contain channels capable of transporting at least one macromolecule across the length of channels, in which the macromolecule is in an elongated form. Such channels can have openings large enough to permit the entrance of the ends of the macromolecules. In certain embodiments, it is preferred that such channels also have trench widths and trench depths narrow enough to restrict the movement of the macromolecules to primarily one direction along the surface of the substrate. Preferably such channels are not interconnected.

[0059] In certain preferred embodiment, of the present invention, the nanochannel array is capable of transporting at least one biopolymer across the length of said channels. In these embodiments, the geometry of the channels permits the biopolymers to enter and move along the channels in at least one direction. Preferably, the channel surfaces are treated with a non-sticking agent, as described later, for preventing the adhesion of macromolecules, such as biopolymers, to the inside of the channels.

[0060] In other embodiments, it is preferred that the nanochannel array contains channels capable of transporting at least one unfolded biopolymer across the length of said channels. While not being bound by a particular theory, when the dimensions of the channels are apparently larger than the spatial conformation of the macromolecules, there is at least a partial amount of elongation of the macromolecules in the channels. When the dimensions of the channels are at the same order or below the persistence length, of macromolecules, such as 50 nm for DNA the macromolecules can be sufficiently elongated in an unfolded fashion inside the channels. When the dimensions of the channels fall in between the above-mentioned two scenarios, macromolecules can be partially elongated in these channels. In this case, the macromolecules can be folded, tangled, or both folded and tangled. While it is envisaged that any macromolecule can be transported in an unfolded fashion in the channels of the nanochannel array of the present invention, a variety of suitable unfolded macromolecules include RNA, DMA, denatured unfolded peptide chains, self assembled chemical polymers, and co-polymer chains, and other biopolymers, and combinations thereof.

[0061] In one preferred embodiment, the channel structures of the nanochannel arrays can be formed from linearly adjacent channel walls that span the substrate surface. In other embodiments, the channels can be formed from pillar structures, sell-assembled polymer structures, stacked membrane layers, and nanobeads (particles inside the channels).

[0062] The surface material of the nanochannel arrays can be formed from almost any substrate material, such as a conductive material, a semiconductor material, or a non-conductive material. Examples of conductive materials include metals such as aluminum, gold, silver, and chromium. Examples of semiconductive materials include doped silicon dioxide and gallium arsenide. Examples of non-conductive materials include fused silica, silicon dioxide, silicon nitride, glass, ceramics, and synthetic polymers. The foregoing is exemplarly only.

[0063] In the present invention, the surface of the nanochannel array is typically the surface of the substrate, such as the surface of a silicon wafer. Alternatively, the surface can be a film, such as one adjacently supported by a second substrate. Coating a material on to a second substrate can form films. A suitable coating process includes vapor deposition of a material onto a wafer.

[0064] In certain embodiments of the present invention, the nanochannel array includes at least one optically opaque material layer adjacent to the sealing material. The optically opaque material can be situated between the surface material and the sealing layer, it can be situated inside the channels, it can he situated on top of the sealing, material, or a combination of these. While almost any opaque material that can he deposited as a layer is possible in this embodiment, Aluminum is preferred. For certain embodiments, it is desirable that the opaque layer thicknesses are less then about 50 nm thick. For embodiments containing nanoslits useful for carrying out near-field imaging of the contents of the channels, it is desirable to prepare slits smaller than 50 nm that are etched through the deposited opaque layer but not through the transparent sealing material for maintaining the integrity of the adjacent (underneath) channel. Without being bound by a particular theory, the optically opaque layer functions as a blocking mask for high-resolution near-field excitation. Without being bound to a particular theory, aluminum is particularly preferred as an opaque layer because if has the highest known skin depth of any material at the given wavelength of the excitation light source rendering the smallest thickness of the blocking layer, hence the shortest distance between the slits and the possible target molecules in channels.

[0065] In certain embodiments of the present invention, the nanochannel array has at least one near field slit feature above at least one channel. Such slits should be fabricated as close (less than about 30 nm) to the channel as possible without compromising the integrity of the adjacent sealed channels. The thin wall of sealing material between slit opening and channels could be created by FIB milling or controlled material deposition.

[0066] In a further preferred embodiment, the nanochannel array contains sealing material adjacent to the channel bottom. Sealing material can be provided in channel bottom by depositing a suitable sealing material into the channels prior to or simultaneously with enclosing the channels.

[0067] The nanochannel arrays also preferentially have sealing material adjacent to the channel wall material. In this embodiment, the sealing material can reduce the trench width. This is particularly advantageous for preparing nanochannel arrays from a variety of substrate surfaces that contain channels wider than 150 nm in trench width and deeper than 200 mn in trench depth. In this embodiment, and as described below, sealing material can be deposited into channels by a variety of methods. One suitable method is E-beam evaporation, which creates a point source of material. In E-beam evaporation, a substrate is typically far away from the source compared to the size of the sample, and the angular distribution of the depositing material is very narrow. To achieve a non-uniform deposition the substrate is tilted at a specific angle. The channel walls partially block the deposition of sealing material (like a shadow), and most of the material is be deposited on the channel walls near the upper portion of the channel wall. Beyond a critical depth no deposition will occur as long as the substrate is tilted.

[0068] An alternative and preferred method to provide sealing material in the channels is sputter deposition. In sputter deposition, the sealing material is deposited at all angles, so the instant growth rate at any point on the surface depends on the percentage of the total target area within its line of sight, as is outlined in FIG. 4. Without being bound to a particular theory, sealing material, from a large target scarce 130 that is in close proximity to the substrate surface, can travel along a variety of trajectories (122, 124, 126, 127) and be deposited at different positions in the channels. Sputtering is typically med used because of the divergent nature of the material beam, thus resulting in the faster deposition of target material at the top part of the channels instead of at the bottom of the channels (i.e., surmounting the channel). In time, the sealing material near the top of the channels eventually completely encloses the channel, which prevents further deposition of sealing material into the channel. In one embodiment, the resultant sealing material in the channel results in a profile 128. Suitable sputtering systems are known in the art. A particularly suitable sputtering system has a 200 mm SiO.sub.2 target source which provides high surface coverage and uniformity across 300 mn substrate.

[0069] The lengths of the channels of the nanochannel array can have a wide range. The lengths of the channels can also he the same or different in the nanochannel array. For carrying out macro-molecular analysis using the nanochannel arrays as provided below, it is desirable that he channels are at least 1 millimeter (mm) longer. More typically, the length of the channels is greater than 1 centimeter (cm), and even greater than 5 cm, 15 cm, and 25 cm.

[0070] In another aspect of the present invention there is provided a method of preparing a nanochannel array, which includes the steps of forming a plurality of channels in the material of the surface of a substrate, and depositing a sealing material to surmount the plurality of channels to provide at least substantially enclosed channels. Substrates containing a plurality of channels preferably have a periodicity of 200 nanometers or less, which can be provided by interferometric lithography and nanoimprint lithography techniques, which are disclosed in U.S. Pat. No. 5,772,905, the complete disclosure of which is incorporated by reference herein. As described earlier, various types of materials can be used to prepare surfaces having a plurality of channels. Suitable substrates include semiconductors, dielectrics, polymers, self-assembled biofilm, membranes, metals, alloy, and ceramics.

[0071] The sealing material is preferably deposited to surmount the plurality of channels to render such channels at least substantially enclosed, the substantially enclosed channels having a trench width of less than 150 nanometers and a trench depth of less than 200 nanometers. By "surmount" is meant that the sealing material is preferentially deposited towards the top of the channels compared to the bottom of the channels, resulting in substantially enclosed channels, which is described above and in FIG. 4.

[0072] In certain embodiments of the present invention, the sealing material can be deposited using any of a variety of methods, including chemical vapor deposition, spin coating, film lamination, and thermo-evaporation. Preferably, the sealing material is deposited using electron-beam evaporation or sputtering.

[0073] In certain embodiments of the present invention, sealing material is deposited on substrate surfaces by a sputtering process at gas pressures typically less than about 20 mTorr, more typically less than 10 mTorr, and even more typically less than 5 m Torr. Sputtering is a process of driving molecules off a source target surface (such as SiO.sub.2) using energetic ionic bombardment. Atoms are knocked off from the source target and can be deposited on a variety of substrate surfaces, such as patterned silicon wafers. While not being bound to a particular theory, it is believed that as the gas pressure is reduced, there are fewer particles in the environment of the plasma sputtering chamber, which results in the depositing atoms to travel with fewer collisions before reaching the substrate surface; hence, a more anisotropic and faster deposition. At higher gas pressure such, as 30 mTorr, depositing atoms collide more frequently on their path to the substrate surface, hence a more divergent traveling angles and more isotropic, and slower deposition. At lower gas pressure with more anisotropic and last deposition, more depositing atoms can reach the bottom and lower part of the sidewalk of the trenches, causing relatively faster deposition of scaling material at she bottom and sidewalk comparing to the top of the trenches, this subsequently leads to smaller channel (trenchdimensions.

[0074] The aspect ratio of the trenches being sealed also affects the geometry of the final sealed void space. The higher the depth to width ratio, the less sealing material will be deposited near the bottom of the trench. The lower the depth to width ratio, the smaller and narrower the channel dimensions.

[0075] In one embodiment of carrying out the method of the present invention, at least one reservoir is provided to be in fluid communication with at least one end of at least one of the channels. Channels can be fabricated on the substrate using nanoimprinting and interconnecting structures of pillar arrays. Reservoirs can be defined using photolithography and subsequently pattern transferred to the substrate using reactive ion etching (RIE), chemical etching or FIB milling directly to create reservoirs in fluid communication with the channels. Auxiliary structures, such as nanochannels, for connecting the reservoirs to the channels can also be provided using these methods. Typical depth of the reservoirs and auxiliary structures is typically at least several hundreds of nanometers, preferably at least several micrometers deep.

[0076] In certain embodiments, it is desirable to provide an additional sealing step. A suitable additional sealing step includes application of a planar surface substrate to the top of the sealing material. Alternatively, reservoirs can be formed on the sealing planar substrate. Auxiliary fluid communicating structures larger than about a micron cart also be formed to connect to larger sample reservoir. A variety of schemes to connect reservoirs to the channels can be envisioned: at least 2 reservoirs can be provided in fluid communication with at least 2 separate channels; or at least 10 reservoirs are provided in fluid communication with at least 10 separate channels; or at least 96 reservoirs are provided in fluid communication with at least 96 separate channels; or at least 500 reservoirs are provided in fluid communication with at least 500 separate channels; or at least 5000 reservoirs are provided in fluid communication, with at least 100 separate channels; or combinations thereof.

[0077] In a preferred embodiment of the present invention, the method of preparing the nanochannel arrays is carried out using linear channel array substrates having a periodicity of less than 2.00 nm formed by nanoimprint lithography. In this embodiment, the linear channels have a trench width less than 100 nanometers and a trench depth less than 100 nanometers. In this embodiment, at least a portion of the sealing material is deposited using sputter deposition to provide sealing material adjacent to the channel wall material to narrow the trench width.

[0078] Varying the sealing material deposition parameters is also used to narrow the trench width of the channels. The deposition parameters can be varied to provide trench widths of typically less than 100 nanometers. As more material is deposited, trench widths can be narrowed to less than 75 nanometers, and even less than 50 nanometers, 25 nanometers, and 15 nanometers. Trench widths of about 10 nm can also be provided by the methods of the present invention. Typically, the resulting trench widths after deposition will be greater than 2 nm, and more typically greater than 5 nanometers. Trench depths of less than 175, 150, 125, 100, 75, 50 , and 25 nm can also be provided by the methods of the present invention. Trench depths of about 15 nm can also be provided. Typically, the trench depths will be at least 5 nm, and more typically at least 10 nm.

[0079] In another embodiment of the present invention, the method may also include the step of providing at least one near field slit feature above at least one channel. In this step, the sealing material is typically transparent, such as silicon dioxide, to permit spectroscopic detection of fluorescently labeled macromolecules, such as DNA, inside the channels. This permits the use of optical methods, such as near-field optical imaging, to analyze macromolecules in the channels. Nanochannel arrays suitable for near-field optical analysis can be modified to have nanoslits. As described above, the nanoslit above the channel is thin to permit sufficient evanescent excitation of the fluorescently-labled macro-molecules.

[0080] In one embodiment of the present invention, nanochannel arrays can be prepared having a sufficiently thin seal thickness suitable, for near-filed optical analysis of fluids in the channels beneath the sealing material. In one embodiment, channels having an opaque sealing material thicker than 100 nm can be modified using a suitable fabrication method to provide a nanoslit in the opaque sealing material.

 channels to form at least one of; an insulating layer, a conducting layer, a hydrophilic layer, a hydrophobic layer, or combinations thereof. In this embodiment, fee layer thickness is typically less than half of the trench width.

[0083] In one embodiment, the dimensions, geometry, composition, or combinations thereof of the sealing material adjacent to the walls 114 can be modified and manipulated for corresponding samples being analyzed is the channels, in a particular embodiment. It is desirable to alter the surface properties of the sealing material adjacent to the wall 114 This is carried out by treating at least some of the channels with a surface modifying agent to alter the surfaces interior to said channels.

[0084] In one embodiment, surface-modifying agents are deposited in the channels to improve the transport of macromolecules into and through the channels. Surface-modifying agents are particularly useful where the internal dimensions (trench depthtrench height, or both) are less than about 50 nm. Surface-modifying agents can also reduce or increase hydrophobicity of the surfaces interior to said channels. Nonochannel arrays made according to the present invention can be contacted with solutions containing surface-modifying agents, such as by submerging the nanochannel array into such solutions. Suitable surface-modifying agents include polyethyleneglycol (PEG), surfactants, Bovin Serum Albumin (BSA) protein solution, surface non-specific binding saturation, and anti-protein sticking agents. Application of a pressure differential, such as vacuum, can be used to assist the treatment of the channels. Application of vacuum is also useful for degassing any fluids inside the channels.

[0085] In certain embodiments of the present invention, the surface-modifying agent counteracts the electroosmosis effects inside the channels. While not being bound to a particular theory, the electroosmosis effect is usually due to ionized acidic groups immobilized to the matrix (e.g., attached to the wall) inducing positively charged counter ions in the butler that migrate towards the negative electrodes, causing a bulk flow of liquid that migrates in the direction opposite to that of the negatively charged DNA. Accordingly, reducing electroosmosis effects helps charged macromolecules to be transported into and along the channels.

[0086] In one embodiment of the present invention, the channels can be at least substantially enclosed on the surface of the substrate and substantially open on the edges of the substrate. As described herein, the channels are at least substantially enclosed by controlling the deposition of the sealing material. In one embodiment, the channels are substantially open at the edges, which, are readily provided by cleaving or cutting the substrate to reveal the interior portion of the channels.

[0087] In one embodiment, the deposition of the sealing material completely encloses the plurality of channels, in this embodiment, the sealing layer is at least as thick as the atoms of the sealing material. Typically, the sealing material surmounting the plurality of channels is less than 500 nanometers thick, hi certain embodiments, the sealing material surmounting the plurality of channels can be less than: 100 nm, 50 nm, 25 nm, 10 nm, and 5 nm thick. Typically the sealing, material surmounting fee plurality- of channels is at least 1 nanometer thick, and snore typically at least 2 nm thick. In certain embodiments of the present invention, a step of removing a portion of the sealing material is used to reduce the thickness of the sealing material above at least one channel. Sealing material can be removed by a variety of etching and ebeam deposition methods as further described herein.

[0088] In another aspect of the present invention, there is provided a nanofludic chip that includes a nanochannel array of the present invention. Referring to FIG. 5 the nanofluidic chip 200 has a nanochannel array 100, a substrate 146, and reservoirs 144 for samples and waste (or sample collection). Further provided is FIG. 5 are auxiliary sample ports 140 and auxiliary waste ports for handling fluid sample. The reservoirs are in fluid communication with at least one of the channels, so that the sample reservoirs are capable of releasing a fluid into the channels, and the waste reservoirs are capable of receiving a fluid from the channels. Typically the fluids contain macromolecules for analysis.

[0089] In certain embodiments of the present invention, the nanofludic chip contains at least one sample reservoir is formed in the surface of the substrate. Steps to form reservoirs in nanochannel array substrates are provided above. In this embodiment, at least one waste reservoir in fluid communication with at least one of the channels. Typically, the nanofluidic chip contains at least 1 sample reservoir. A variety of other embodiments include at least 96 reservoirs, and even at least 1000 reservoirs in the nanofluidic chip.

[0090] For use in macromolecular analysis, it is preferred that the nanofludic chip provides at least a portion, of the nanochannel array capable of being imaged with a two-dimensional detector. Imaging of the array is provided by presenting the scaling material face of the nanochannel array to suitable apparatus for the collection of emitted signals, such as optical elements for the collection of light from the nanochannel array. In this embodiment, the nanofluidic chip is capable of transporting a plurality of elongated macromolecules from a sample reservoir and across the channels.

[0091] Is certain embodiments of the present invention, the nanofluidic chip contains an apparatus for transporting macromolecules from the sample reservoirs, through the channels, and into the waste reservoirs. A suitable apparatus includes at least one pair of electrodes capable of applying an electric field across at least some of the channels in at least one direction. Electrode metal contacts can be integrated using standard integrated circuit fabrication technology to be in contact with at least one sample and at least one collection/waste reservoir to establish directional electric field. Alternating current (AC), direct current (DC), or both types of fields can be applied. The electrodes can be made of almost any metal, and are typically thin Al/Au metal layers deposited on defined line paths. Typically at least one end of one electrode is in contact with buffer solution in the reservoir.

[0092] In certain embodiments of the present invention, the nanofluidic chip contains at least two pair of electrodes, each providing an electric field in different directions. In this embodiment, adjacent clusters of channels connect individual isolated reservoir. With at least two sets of independent electrodes, field contacts can be used to independently modulate the direction and amplitudes of the electric fields to move macromolecules at desired speed or directions.

[0093] In another aspect of the present invention, there is provided a system (FIG. 10, 300) that is suitable for carrying out macromolecular analysis. In the present invention, the system includes a nanofluidic chip as described herein, and an apparatus for detecting at least one signal transmitted from one or more fluids in the nanochannel array of the nanofludic chip.

[0094] In various embodiments of the present invention, the system further includes at least one of the following: a transporting apparatus to transport a fluid through at least one of the channels; a sample loading apparatus for loading at least one fluid to the sample reservoirs in the nanofludic chip; and a data processor. The various components of the system 300 are connected together, and the general principles of operation are illustrated in FIG. 10.

[0095] The nanofludic chip 300 used in the system is typically disposable, individually packaged, and having a sample loading capacity of 1-50,000 individual fluid samples. The nanofluidic chip typically has at least one interconnecting sample delivery microchannel to provide fluid samples into the channels, as well as sample loading openings and a reservoir, or sample loading openings and plates connected with a sealing mechanism, such as an O-ring. Metal contacts for connecting the electrodes 292 and an electric potential generator 216 are also provided in the nanofluidic chips. Suitable metal contacts can be external contact patches that can be connected to an external scanning/imaging/electric-field tuner.

[0096] The nanofluidic chip is preferably encased in a suitable housing, such as plastic, to provide a convenient and commercially-ready cartridge or cassette. Typically the nanofluidic cartridges will have suitable features on or in the housing for inserting, guiding, and aligning the sample leading device with the reservoirs. Insertion slots, tracks, or both can be provided in the plastic case.

[0097] Macromolecular fluid samples that can be analyzed by the system includes fluids from a mammal, (e.g., DMA, cells, blood, biopsy tissues), synthetic macromolecules such as polymers, and materials found in nature (e.g., materials derived from plants, animals, and other life forms). Such said samples can be managed, loaded, and injected using automated or manual sample loading apparatus of the present invention.

[0098] In one embodiment of the present invention, the system includes an apparatus to excite the macromolecules inside the channels and detect and collect the resulting signals. A suitable apparatus is illustrated in FIG. 10; a laser beam 204 is focused using a focusing lens 206 to a spot on the nanochannel array 100. The generated light signal from the macromolecules inside the channels (not shown) is collected by a collection lens 208, and reflected off a dichroic mirror 218 into an optical path 220, which is fed into a CCD (charge coupled device) camera. Various optical components and devices can also be used in the system to detect optical signals, such as digital cameras, PMTs (photomultlplier tubes), and APDs (Avalanche photodiodes.

[0099] In another embodiment of the present invention, the system includes a data processor. The data processor can be used to process the signals from the CCD to project the digital image of the nanochannel array on a display 212. The data processor can also analyze the digital image to provide characterization information, such as macromolecular size statistics, histograms, karyotypes, mapping, diagnostics information and display the information in suitable form for data readout 214.

[0100] In another aspect of the present invention, there is provided a method of analyzing at least one macromolecule. In this invention, the analysis includes the steps of providing a nanofluidic chip according to the present invention, providing the at least one sample reservoir with at least one fluid, said fluid comprising at least one macromolecule; transporting the at least one macromolecule into the at least one channel to elongate said at least one macromolecule; detecting at least, one signal transmitted horn the at least one elongated macromolecule; and correlating the detected signal to at least one property of the at least one macromolecule.

[0101] In one embodiment of the present invention, the method of analyzing a macromolecule includes wetting the channels rising capillary action with a buffer solution or a buffer solution containing macromolecules. Macromolecules such as polymers and DNA can introduced into nanochannel arrays by electric field.


[0104] Macromolecule fluid samples can be loaded through reservoirs in the nanofluidics chip and transported via interconnecting microchannels. The macromolecules are partially elongated before one end of the macromolecule enters the channels; they are substantially fully elongated when completely inside the channels. The fluorescent signals can be excited by the appropriate excitation sources and emission signals can be collected via imaging camera or detectors, in a linear scanning mode or CCD image integration. The signals collected can be analyzed by data processing software and user-defined major parameters (intensity/photons, major axis, minor axis, background signal)can be recorded and measured.

 [0106] The typical concentration of the macromolecules in the fluid will be one macromolecule, or about at least attogram per ml, more typically at least one femtogram per ml, more typically at least one picogram per ml, and even more typically at least one nanogram per ml. Concentrations will typically be less than 5 micrograms per milliliter and more typically less than 0.5 micrograms per milliliter.


[0111] After NIL and etching, non-uniform deposition of sealing material was provided by e-beam evaporation with a tilted sample wafer at various angles or sputter deposition using a large source target. This step was used to both reduce the trench width and seal the channels.


[0113] In the following examples, nanochannel arrays were prepared using a process to deposit SiO.sub.2 sealing material, on patterned substrates by sputtering. Channel openings were prepared, by cleaving the substrate and imaged by Scanning Electronic Microscope (SEM). Results are as follows and shows mat the trench widths are narrowed by the deposition of the sealing material using sputtering:

Example 1

[0114] A 100 mm silicon substrate was provided having a plurality of parallel linear channels that had an 85 nm trench width and a 200 nm trench height (FIG. 7a). This substrate was sputtered at a gas pressure of 5 mTorr according to the general procedures given above. After sputtering, the channels had a 52 nm trench width, a 186 nm, trench height, and a seal thickness of 200 nm (FIG. 7b). Apparently, the trench height increased slightly as a result of the sealing process forming a cone-shaped seal shove the channel.

Example 2

[0115] A patterned substrate having a 65 nm trench width and about 100 nm trench height prior to sputtering formed a nanochannel array having a 17 nm trench width, a 68 nm bench height, and a channel seal thickness of about 350 nm. Sputtering gas pressure was 5 mTorr.

Example 3

[0116] A patterned substrate having a 50 nm trench width and about 80 nm trench depth prior to spattering formed a nanochannel array having a 10 nm trench width, 51 nm trench height, and a channel seal thickness of 350 nm. Sputtering gas pressure was 5 mTorr.

Example 4

[0117] A substrate containing a two-dimensional array of pillars was made using a two-step NIL process with the channel amid rotated 90.degree. between the imprinting steps (FIG. 6a). The pillar array structure, is subsequently completely sealed with silicon dioxide using a 29 minute deposition time. The seal thickness was about 500 nm. A profile view of the channel is depleted in FIG. 6b and a top view of the completely sealed nanochannel array is depicted in FIG. 6d.

Example 5

[0118] Example 4 was repeated except the sputter deposition time was 17 minutes to provide a nanochannel array that is not completely sealed. A top view scanning electron micrograph of this nanochannel array is provided In FIG. 6d. The seal thickness was 300 nm.

Example 6

[0119] A nanoslit is provided in a channel prepared with a silicon dioxide sealing material for carrying out near-field analysis saving a seal thickness greater than about 100 nm is modified by using FIB to create a nanoslit having a thicknessless than 100 nm. FIG. 8 shows a schematic of how the deposited sealing material on a nanochannel array is first milled away using FIB from the sealing material situated above the sealed channels. Subsequently, aluminum is deposited to create an opaque layer to provide optical contrast at the slit.

Example 7

[0120] This example shows bow a nanochannel array can be prepared from a substrate having a plurality channels larger than 150 nm wide by 150 nm deep. A substrate is prepared by photolithography techniques to provide a plurality of channels with width of greater than 1.5 micron using conventional optical lithography techniques. Contact aligner such as Karl Suss MA-6 to provide a pattern resolution at low micron level; Industrial projection stepper. The angle of the incident depositing beam of sealing material is varied to reduce the trench width and height to less than 150 nm and 150 nm, respectively, and to substantially seal by providing shallow tangential deposition angles.

Example 8

[0121] This example provides a nanochannel array using an e-beam, technique. A substrate is provided, as in Example 1. Silicon dioxide is deposited by an e-beam (thermo) evaporator (Temescal BJD-1800) onto the substrate. The substrate is placed at various angles incident to the depositing beam from the silicon dioxide source target; the deposition rate is set to about 3 nm/minute and 150 nm of sealing material is deposited in about 50 minutes.

Example 9

[0122] In this example, a nanochannel array is contacted with a surface-modifying agent. A nanochannel array made according to Example 1 is submerged in a surface-modifying agents solutions containing polyethylene glycol inside a vacuum chamber overnight to facilitate wetting and treatment of the channels and degas the air bubbles that might be trapped inside the channels.

Example 10

[0123] This example shows the preparation of a nanochannel array having a metal sealing material. An e-beam (thermo) evaporator (Temescal BJD-1800) was used to deposit Chromium (Cr) onto a nanochannel array chip (trench width 80 nm, trench depth 80 nm, SiO2/Si substrate). The substrate was placed at various angles to the incident depositing beam from the source target, the deposition rate was set at 2.0-3.6 nm/minute. The resulting trench width was 20 nm, trench depth less than 80 nm, and the channels were substantially closed.

Example 11

[0124] This example shows the process of adding an optically opaque layer to a nanochannel array. A nanochannel array made according to Example 3 is placed perpendicular to the incident depositing beam, to provide an opaque layer less than 50 nm thick. An aluminum source target is selected for depositing on top of the SiO2 sealing material above the sealed channels. The deposition rate was set at 2.0-3.6 nm/minute.

Example 12

[0125] This example describes the steps needed to provide a near-field slit in a nanochannel array. FIB was used to mill narrow silts less than 50 nm in width in the direction, perpendicular to the long axis of the nanochannel array of Example 11. The depth of the FIB milling was controlled to expose the underlying thin SiO2 sealing material above the nanochannel array.

Example 13

[0126] This example describes how to provide a sample reservoir with a nanochannel array to provide a nanofludic chip. A nanochannel array having 100 nm wide, 100 nm deep channels was made according to general procedures of Example 1. The nanochannel array was spin-coated with a photoresist and imaged with a photomask, to provide regions on opposite ends of the nanochannel array. The exposed areas were etched using reactive ion etching to expose the channel ends and to provide a micron-deep, reservoir about a millimeter wide on the opposite ends of the channels at the edge of the substrate.

Example 14

[0127] This example describes how to fill a nanofluidic chip with a fluid containing DNA macromolecules to analyze the DNA. A cylindrical-shaped plastic sample-delivery tube of 2 mm diameter was placed in fluid communication with one of the reservoirs of the nanochannel array of Example 13. The delivery tube is connected to an external sample delivery/collection device, which is in torn connected to a pressure/vacuum generating apparatus. The channels are wetted using capillary action with a buffer solution. A buffer solution containing stained lambda phage macromolecules (lambda DMA) were introduced into the nanochannel array by electric field (at 1-50 V/cm); the solution concentration was 5 microgram/mL and the lambda DMA was stained at a ratio of 10:1 base pair/dye with the dye TOTO-1 (Molecular Probes, Eugene, Org.). This solution of stained DNA was diluted to 0.1-0.5 microgram/mL into 0.5.times.TBE (tris-boroacetate buffer at pH 7.0) containing 0.1M of an anti-oxidant and 0.1% of a linear polyacrylamide used as an anti-sticking agent.

Example 14

[0128] A nanofluidic chip made according to Example 12, having channel dimensions of 100 nm.times.100 nm was filled using capillary action with a buffer solution containing stained genomic DNA to draw the DNA macromolecules into the channels with an electric field. Bacteria phage DNA molecules Lambda (48.5 kb) and T4 (168.9 kb) were stained with the dye TOTO-1 and BOBO-3 respectively. This solution of stained DNA was diluted to 0.5 .mu.g/mL into 0.5 .times. TBE containing 0.1 M dithlothreatol as an anti-oxidant and 0.1% of a linear acrylamide used as an anti-sticking agent). A Nikon Eclipse TE-300 inverted microscope with a 60.times. (N.A. 1.4) oil immersion objective was used with an Ar:K laser (Coherent Lasers) as an excitation source at 488 nm and 570 nm. A Roper Scientific Peotamax intensified cooled CCD camera with a 512.times.512 pixel array and 16 bits digital output-was used to image the molecules. Digital Image was analyzed using a data processor by NTH Image software. FIG. 9b shows an integrated image of the stretched Lambda and T4 phage genomes side by side in the channels. The inset of 9b shows the near perfect linear fit of the directly measured length obtained from the digital image plotted against their genome size, (R2 is 0.99996). FIG. 9c shows an army of fluorescently-labled genomic DNA molecules aligned and stretched in the channels with the size ranging from 10 kb to 194 kb. This shows that millions of center-meter long parallel channels could be fabricated over the whole wafer. Accordingly, the entire length of genomic DNA molecules can be stretched and analyzed.

Example 15

[0129] Example 14 is repeated, but with a 96 multiple reservoir system. A nanofludic chip made according to Example 12 is modified with a photomask to provide 96 sample reservoirs, each reservoir connected to 1000 channels along one edge of the 100 mm substrate. 96 different DMA samples are delivered and injected using capillary fibers connected to the sample reservoirs. 96 collection reservoirs are connected to the corresponding ends of the channels to collect the DNA samples.

Example 16

[0130] This example describes a system used for carrying out analysis of macromolecules. The system contains an automated 96-caplilary auto-injection sample loader to deliver 96 macromolecular fluid samples into the delivery ports of a nanofluidic cartridge. The nanofluidic cartridge is a nanofluidic chip encased by a plastic polycarbonate housing, having delivery ports and collection ports for connection to microcapillary tubing, and embedded metal contacts for connection to electrodes on the nanofluidic chip. The cartridge can be inserted in a cartridge holder, which is integrated with an a laser excitation source and suitable optical components to provide the excitation of and collection of optical signals emanating from sample fluids within the nanochannel arrays of the nanofluidic chip. The signal detection/collection apparatus is a cooled CCD digital camera. Signals from the digital camera are analyzed by a data processor using NIH image analysis software, and displayed on a monitor.

Example 17

[0131] This example describes how to use the system of Example 16 to size one DNA macromolecule. A single Anthrax spore is lysed to extract its entire genomic contents (DNA) with 10 microliters of a buffer solution and stained with fluorescent dyes. The sample loader is inserted into the delivery ports of the cartridge and injects the DNA-containing fluid. The electrodes are activated and the DNA macromolecules are transported into the nanochannel array, where they become elongated. The fluorescent steins on die DMA are excited by the excitation source, and their emission signals are collected using the CCD cameras. The signals collected, analyzed and recorded for intensity and position by the data processor. The length of a single DMA is detected and intensity profile is plotted.

[0132] In another aspect of the present invention, there is provided 144. A cartridge comprising at least one nanofluidic chip, said cartridge capable of being inserted and removed from a system for carrying out macromolecular analysis, said at least one nanofluidic chip comprising at least one nanonanochannel array, said nanonanochannel array comprising.

[0133] a surface having a plurality of channels in the material of the surface, said channels having a trench width of less than about 150 nanometers and a trench depth of less than 200 nanometers;

[0134] at least some of the channels being surmounted by sealing material to render such channels at least substantially enclosed. 


Cruel Reality – Lawyers are messing with science on intellectual property affairs as they have no handle on it.
So, collusion with USPTO is unleashing a wave of flawed patents like this one more below:


Scientific Flaw.

The ‘wick’ 34 cannot work below the water table reference.
The device 34 is just a porous drain and never a wick.
Wick is a device on oil lamps that takes fuel toward the flame inside the Unsaturated Hydraulic Zone (Negative Pressure).
Lay people employing wicking terminology does not understand oil lamps and the functioning of wicks.

My Demand to USPTO

It is not that hard to argue at the Court of the Law that Hydrological issues should be examined by Hydrologists.
My demand to USPTO is the same as the first letter sent on Oct. 2006:

1. Hire Examiners with background in Hydrogeology and/or Soil Physics so that they have full comprehension of fluids moving on porosity;
2. Cancel issued patents with scientific flaws. Obsolete patents are cancelled naturally by becoming outdated;
3. Make a public statement about Hydrology negligence hurting all Hydrological community as well as my project that needs experts in Hydrology to protect the content of my issued claims.
4. Compensate for my losses since as an inventor filing patents I was not expecting lay people handling hydrology in the examination process by USPTO.
5. Since USPTO is failing to protect my IP rights my patents should be eligible for time extension of their expiration dates (new demand).
6. Issue a bill requiring Hydrology be handled by Hydrologists preventing laypeople from harming standing common knowledge in the scientific literature (new demand).
7. Make people accountable for breaking the Law regarding my complaints (new demand).
_______________________________________________________________________________

Understanding the bias – The size of the Hydrological Gap

Wick/wicking is not a word found on my Hydrology textbooks but it is in the patent classification system at USPTO guiding lay inventors and lay Patent Examiners pretending they are known in the art.

In 1856 Henry Darcy proposed an equation Law to address fluids moving on porosity for Hydraulic Conductivity. Afterwards in 1907 Edward Buckingham suggested a change to address negative pressure flow for Unsaturated Hydraulic Flow (wick/wicking).

Today May 12, 2014 searching at USPTO:

Thermal/Heat Conductivity is mentioned in                                                                96,025 issued patents
Electrical/Electric Conductivity is mentioned in                                                          72,719 issued patents
Saturated Hydraulic Conductivity mentioned in only                                                        702 issued patents
Hydraulic Conductivity (wicking for lay people) is mentioned in only                                     27 issued patents
Wick/wicking (Unsaturated Flow if not flawed) is mentioned in                                    32,206 issued patents

_______________________________________________________________________________

Abraham Duncan Stroock
Dept: Chemical and Biomolecular Engineering
Title: Associate Professor

Education

Ph.D., Harvard University, Chemical Physic (2002)
M.S., University Paris VI and XI,Solid State Physics (1997)
B.A. Cornell UniversityPhysics (1995)


De: Abraham Duncan Stroock [mailto:abe.stroock@cornell.edu]
Enviada em: terça-feira, 22 de abril de 2014 22:20
Para: Elson Silva, PhD
Assunto: RE: [06856] Protecting Hydrology Science from REINVENTION by corrupt LAY PEOPLE colluding with USPTO - US Pat 8,701,469

Dear Dr. Silva,
‘...  If you have a point to make about my treatment of hydrological concepts, I ask that you take the time to explain your specific points of disagreement.  I note that my work is better represented in my publications (available at http://www.stroockgroup.org/home/publicationsthan in patentsas the lawyers have been translated the latter into legalese that I do not understand.

Best regards,

Abe
___________________________________________________________
De: Elson Silva, PhD [mailto:el_silva@uol.com.br]
Enviada em: terça-feira, 22 de abril de 2014 23:40
Para: 'Abraham Duncan Stroock'
Cc: cko3@cornell.edu; TDO1@cornell.edu; MGS22@cornell.edu; SBW11@cornell.edu; el_silva@uol.com.br
Assunto: RES: [06856] Protecting Hydrology Science from REINVENTION by corrupt LAY PEOPLE colluding with USPTO - US Pat 8,701,469
Prioridade: Alta

Abe,

You are so naive.

‘…Are you sure you got your PhD at Harvard? ‘

Lawyers learn nothing about Hydrology in Law School.

As far as I know no Law School provides Hydrology teaching . . . No Lawyer could discuss Hydrology having no expertise in the subject!

This is funny!
You do not give your scientific papers to Lawyers, so why are your patents different?
(By the way, was it a Lawyer who wrote your PhD thesis?)
Also, Lawyers are illiterate on the functioning of science, besides most scientists have no idea about Epistemology, Metaphysics, Logics, and History of Science (Philosophy of Science).

USPTO is a sham system allowing reinvention sometimes by flawed lazy patents from wealthy parties letting Lawyers step over boundaries beyond their background in Law Schools….’


I would like to invite people around the world to send messages to those 14,302 emails below pledging them to be honest and stop violating intellectual property rights allowing huge corporations and UNIVERSITIES to reinvent issued patents shamefully dishonoring HYDROLOGY SCIENCE and breaking the Law.


0756353@princeton.edu
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