# US 11,994,507 B2 · Nanopore sensor calibration and operation with a fluidic passage > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,994,507 B2 - Document ID: US11994507B2 - Status: Granted - Kind: B2 - Application number: 17/007,290 - Filed: 2020-08-31 - Published: 2020-12-24 - Granted: 2024-05-28 - Earliest claimed: 2015-02-05 - Assignees: President and Fellows of Harvard College - Applicants: President and Fellows of Harvard College - Inventors: Ping Xie - Web page: https://patentgrep.com/patent/US11994507B2 ## CPC - G01N 33/48721: {Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68)} - B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces} - B01L 3/50273: {characterised by the means or forces applied to move the fluids} - C12Q 1/6869: Methods for sequencing - G01N 27/4145: {specially adapted for biomolecules, e.g. gate electrode with immobilised receptors} - G01N 27/4146: {involving nanosized elements, e.g. nanotubes, nanowires} - G01N 27/4163: {checking the operation of, or calibrating, the measuring apparatus (G01N27/3274, G01N27/4175 and G01N33/0006 take precedence)} - G01N 27/44726: {using specific dyes, markers or binding molecules} - G01N 27/4473: {by electric means} - G01N 27/44791: {Microapparatus (sample containers with integrated microfluidic structures B01L3/5027)} - B01L 2300/0645: Electrodes - B01L 2300/0867: Multiple inlets and one sample wells, e.g. mixing, dilution - B01L 2300/0896: Nanoscaled - B01L 2300/12: Specific details about materials - B01L 2400/0421: electrophoretic flow - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract A first fluidic solution having a first ionic concentration is provided in a first fluidic reservoir in direct fluidic connection with a nanopore. A second fluidic solution, having a second ionic concentration, is provided in a second fluidic reservoir disposed in fluidic connection with the nanopore through a fluidic passage having at least one fluidic section in which the section length is greater than the section width. The electrical potential local to the fluidic passage is measured, and the resistance of both the fluidic passage the nanopore are determined based on the fluidic passage electrical potential. The fluidic passage resistance is compared with the nanopore resistance and at least one of the first and second ionic concentrations is adjusted based on the comparison. The measuring, determining, comparing, and adjusting steps are conducted until the fluidic passage resistance is between about 0.1 times and about 10 times the nanopore resistance. ## Claims ### Claim 1 A method for calibrating a nanopore sensor comprising: providing a first fluidic solution, having a first ionic concentration, in a first fluidic reservoir, said first fluidic reservoir being disposed in fluidic connection with an inlet to a nanopore; providing a second fluidic solution, having a second ionic concentration, in a second fluidic reservoir, said second fluidic reservoir being disposed in fluidic connection with an outlet of said nanopore through a fluidic passage having at least one fluidic section with a fluidic section length greater than a fluidic section width and a passage width greater than a diameter of the nanopore; measuring an electrical potential local to the fluidic passage; determining a fluidic passage fluidic resistance, RFP, and a nanopore fluidic resistance, RPore based on measured fluidic passage electrical potential; comparing the fluidic passage fluidic resistance, RFP, with the nanopore fluidic resistance, RPore; adjusting at least one of the first ionic concentration and the second ionic concentration based on said comparing; and carrying out the measuring, determining, comparing, and adjusting steps until the fluidic passage fluidic resistance, RFP, is between about 0.1 times and about 10 times the nanopore fluidic resistance, RPore. ### Claim 2 (depends on 1) The method of claim 1 wherein adjusting at least one of the first ionic concentration and the second ionic concentration comprises adjusting at least one of the first ionic concentration and the second ionic concentration until one of the first ionic concentration and the second ionic concentration is at least about one hundred times greater than the other of the first ionic concentration and the second ionic concentration. ### Claim 3 (depends on 1) The method of claim 1 wherein adjusting at least one of the first ionic concentration and the second ionic concentration comprises adjusting said first ionic concentration until said first ionic concentration is at least about one hundred times greater than said second ionic concentration. ### Claim 4 (depends on 1) The method of claim 1 wherein measuring an electrical potential local to the fluidic passage comprises measuring a signal produced by at least one electrical transduction element disposed in the nanopore sensor at a site that produces a signal indicative of electrical potential local to the fluidic passage. ### Claim 5 (depends on 1) The method of claim 1 wherein measuring an electrical potential local to the fluidic passage comprises measuring a signal produced by an electrical transduction element disposed at in the fluidic passage. ### Claim 6 (depends on 1) The method of claim 1 wherein measuring an electrical potential local to the fluidic passage comprises measuring a signal produced by an electrical circuit connected to an electrical transduction element disposed at in the fluidic passage. ### Claim 7 (depends on 1) The method of claim 1 wherein measuring an electrical potential local to the fluidic passage comprises measuring a signal produced by a transistor. ### Claim 8 (depends on 1) The method of claim 1 wherein measuring an electrical potential local to the fluidic passage comprises measuring a voltage of a sensing element disposed at the fluidic passage. ### Claim 9 (depends on 1) The method of claim 1 further comprising operating the nanopore sensor, said operation comprising: providing in said first fluidic solution at least one molecule; translocating said at least one molecule from said first fluidic solution to said second fluidic solution through said nanopore and said fluidic passage; and sensing a translocation electrical potential local to the fluidic passage during the translocating of said at least one molecule. ### Claim 10 (depends on 9) The method of claim 9 wherein translocating said at least one molecule through the nanopore comprises translocating at least one molecule selected from DNA, DNA fragments, RNA, RNA fragments, PNA, nucleotides, nucleosides, oligonucleotides, proteins, polypeptides, amino acids and polymers. ### Claim 11 (depends on 9) The method of claim 9 further comprising producing a signal indicative of said at least one molecule translocation. ### Claim 12 (depends on 9) The method of claim 9 further comprising applying between the first ionic solution and the second ionic solution an electrical voltage operative to electrophoretically translocate said at least one molecule through said nanopore. ### Claim 13 (depends on 9) The method of claim 9 wherein translocating at least one molecule through said nanopore comprises translocating at least one molecule through a protein nanopore. ### Claim 14 (depends on 9) The method of claim 9 further comprising producing an electrical signal indicative of said least one molecule translocation. ### Claim 15 (depends on 14) The method of claim 14 further comprising processing said electrical signal indicative of molecule translocation as a function of time to determine duration of translocation of said at least one molecule through the nanopore. ### Claim 16 (depends on 14) The method of claim 14 further comprising processing said electrical signal indicative of molecule translocation as a function of time to identify said at least one molecule. ### Claim 17 (depends on 9) The method of claim 9 further comprising controlling rate of molecule translocation with a molecular motor disposed at the nanopore. ### Claim 18 (depends on 9) The method of claim 9 wherein translocating at least one molecule through the nanopore and the fluidic passage comprises translocating at least one molecule through a lateral fluidic passage section and through a vertical fluidic passage section of said fluidic passage. ### Claim 19 (depends on 9) The method of claim 9 wherein translocating at least one molecule through the nanopore and the fluidic passage comprises translocating at least one molecule through a lateral fluidic passage section of said fluidic passage, the lateral fluidic passage section being disposed in a surface layer on a nanopore support structure. ### Claim 20 (depends on 9) The method of claim 9 wherein translocating at least one molecule through the nanopore and the fluidic passage comprises translocating at least one molecule through a nanopore that is vertical and translating the molecule through at least one fluidic passage section that is horizontal relative to the vertical nanopore.