# US 10,309,958 B2 · Method and apparatus for bacterial monitoring > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,309,958 B2 - Document ID: US10309958B2 - Status: Granted - Kind: B2 - Application number: 14/779,977 - Filed: 2014-03-25 - Published: 2016-03-03 - Granted: 2019-06-04 - Earliest claimed: 2013-03-25 - Assignees: Yissum Research Development Company of the Hebrew University of Jerusalem; Technion Research & Development Foundation Limited - Applicants: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd.; Technion Research & Development Foundation Limited - Inventors: Amir Sa'ar; Ester Segal - Web page: https://patentgrep.com/patent/US10309958B2 ## CPC - G01N 33/5302: {Apparatus specially adapted for immunological test procedures} - G01D 5/266: {by interferometric means (G01D5/353 takes precedence)} - G01N 21/4738: {Diffuse reflection (precedence is given to G01N21/55 - G01N21/57 if specular component is taken into consideration), e.g. also for testing fluids, fibrous materials} - G01N 21/4788: {Diffraction (for sizing particles G01N15/0205)} - G01N 21/85: Investigating moving fluids or granular solids - G01N 2201/0446: Multicell plate, sequential - G01N 2201/061: Sources - 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) - G01D: MEASURING NOT SPECIALLY ADAPTED FOR A SPECIFIC VARIABLE; ARRANGEMENTS FOR MEASURING TWO OR MORE VARIABLES NOT COVERED IN A SINGLE OTHER SUBCLASS; TARIFF METERING APPARATUS; MEASURING OR TESTING NOT OTHERWISE PROVIDED FOR ## Abstract A system for detecting target elements such as bacteria in a host analyte, comprising a substrate with an ordered array of wells having diameters to fit the size of the targets. The substrate may be a periodic macro-PSi array structure (MPSiAS) illuminated with a broadband source. The reflected light spectrum diffracted from the substrate is optically analyzed to provide the effective optical depth of the wells. Fast Fourier Transform analysis may be used for the optical analysis. Entry of target elements into wells is detected by the change in the effective optical depths of the wells. Micro-organisms as large as bacteria and viruses having dimensions comparable with the wavelength of the illumination can thus be detected. Wells with an inner section impenetrable by the target cells enables compensation for environmental changes. The detection may be performed in real time, such that production line bacterial monitoring may be achieved. ## Claims ### Claim 1 A system for detecting target elements in a host analyte, said system comprising: a substrate containing an ordered array comprising a predetermined pattern of wells formed in its surface, at least some of said wells having predetermined lateral dimensions such that said target elements can fit therein; a broadband illumination source configured to illuminate said surface of said substrate with broadband illumination over a range of wavelengths; an optical detector disposed such that it collects illumination diffracted from said substrate, and outputs a reflected spectrum signal; and a signal processing unit, adapted to analyze said reflected spectrum signal to provide a measure of the effective optical depth of said wells, wherein at least some of said wells have a lateral dimension at least as large as those wavelengths of said illumination source which are diffracted from said substrate and detected by said optical detector, to output said reflected spectrum signal which is analyzed by said signal processing unit, such that said system can detect said target elements. ### Claim 2 (depends on 1) A system according to claim 1 wherein said signal processing unit analyzes said reflected spectrum signal using the Fast Fourier Transform. ### Claim 3 (depends on 1) A system according to claim 1 wherein said optical detector is disposed normal to said substrate, such that it collects diffracted light of zero order from said substrate. ### Claim 4 (depends on 1) A system according to claim 1, wherein said effective optical depth of said wells provides an indication of the concentration of said target elements captured within said array of wells. ### Claim 5 (depends on 2) A system according to claim 2, wherein said effective optical depth is determined from the position of a peak obtained from the results of Fast Fourier transform analysis on said reflected spectrum. ### Claim 6 (depends on 1) A system according to claim 1, wherein said ordered array of wells comprises a lamellar photonic crystal grating. ### Claim 7 (depends on 1) A system according to claim 1, wherein said substrate is a silicon chip, and said ordered array of wells are constructed by microelectronic fabrication processes. ### Claim 8 (depends on 1) A system according to claim 1, wherein said target elements are bacterial cells having dimensions larger than the wavelengths of said illumination source detected by said optical detector element. ### Claim 9 (depends on 1) A system according to claim 1, wherein said wells comprise capture probes having a high affinity to the target elements intended to be measured by said system. ### Claim 10 (depends on 9) A system according to claim 9, wherein said target elements are micro-organisms, and said capture probes are any one of antibodies, aptamers or other peptides. ### Claim 11 (depends on 10) A system according to claim 10, wherein said microorganisms are bacterial cells, and said capture probes are specific antibodies. ### Claim 12 (depends on 1) A system according to claim 1, wherein said system is adapted to provide real time detection of microorganisms. ### Claim 13 (depends on 9) A system according to claim 9, further comprising a cell nutrient supply, such that the growth of a microorganism can be observed after application of said nutrient supply. ### Claim 14 (depends on 1) A system according to claim 1 wherein at least some of said wells include a recognition moiety adapted to said target elements to be detected. ### Claim 15 (depends on 14) A system according to claim 14, wherein said substrate comprises at least two different regions, the wells in each of said regions including a different recognition moiety, such that each of said different regions can detect different target elements concurrently. ### Claim 16 (depends on 1) A system according to claim 1, wherein at least some of said wells have at least two sequential sections having different lateral dimensions, and wherein a second section, further from said surface than a first section, has a lateral dimension less than that of said first section. ### Claim 17 (depends on 16) A system according to claim 16, wherein the dimensions of said second section are such that said target elements cannot penetrate said second section, while said host analyte can. ### Claim 18 (depends on 16) A system according to claim 16, wherein change of said measured effective optical depth of said second section of at least some wells is utilized as a marker to compensate for changes in environmental conditions that cause said effective optical depth of both of said first and second sections to change. ### Claim 19 (depends on 16) A system according to claim 16, wherein said second section is provided with sensitivity to a material which targets trapped in said first section may secrete. ### Claim 20 (depends on 19) A system according to claim 19 wherein said sensitivity provides further information regarding the level of targets trapped in said first sections of said wells. ### Claim 21 (depends on 1) A system according to claim 1 wherein at least some of said wells have a depth at least several times as large as the wavelengths of said illumination source detected by said optical detector. ### Claim 22 (depends on 1) A system according to claim 1 wherein said wells in said predetermined pattern are arranged in an ordered array of rows and columns.