# US 2019/0170748 A1 · Antibiotic susceptibility testing via plasmonic imaging and tracking > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0170748 A1 - Document ID: US20190170748A1 - Status: Published application - Kind: A1 - Application number: 16/270,312 - Filed: 2019-02-07 - Published: 2019-06-06 - Earliest claimed: 2015-08-03 - Assignees: Arizona Board of Regents on behalf of Arizona State University - Inventors: Nongjian Tao; Karan Syal - Web page: https://patentgrep.com/patent/US20190170748A1 ## CPC - G01N 33/56916: {Enterobacteria, e.g. shigella, salmonella, klebsiella, serratia} - G01N 33/54373: {involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings} - G01N 33/56911: {Bacteria} - 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) ## Abstract A rapid antibiotic susceptibility test (AST) based on the detection and quantification of the movement of single bacterial cells with a plasmonic imaging and tracking (PIT) technology. The PIT-based AST detects changes in the metabolic activity of the bacterial cells long before cell replication, and allows rapid AST for both cultivable and non-cultivable strains. PIT tracks 3D movement with sub-nanometer resolution and millisecond temporal resolution. PIT also allows simultaneous measurement of the binding kinetic constants of antibiotics and bacterial metabolic state after the introduction of antibiotics. ## Claims ### Claim 1 A system for antibiotic susceptibility testing using plasmonic imaging for bacterial cells comprising: a computer processor; a plasmonic imaging and tracking (PIT) system coupled to the computer processor, the PIT including an inverted microscope lens, a light source located to impinge on the inverted microscope lens, a slide, located to receive light from the microscope lens, the slide having a metallic coated surface, a mirror, located to receive reflected light from the metallic coated surface through the inverted microscope lens and transmit the reflected light, and a detector located to receive the reflected light from the mirror; where the metallic coated surface has tethering molecules attached; bacteria populating the metallic coated surface; a device for adding an antibiotic to the metallic coated surface after it has been populated with the bacteria; where the computer processor includes and is programmed to execute a computer program for tracking a first set of 3D motion changes of the bacteria before addition of the antibiotic, then tracking a second set of 3D motion values of the bacteria in the presence of the antibiotic; and then comparing the first and second 3D motion values to determine changes in the 3D motion of the bacteria after addition of the antibiotic. ### Claim 2 (depends on 1) The system of claim 1 wherein the tethering molecules comprise tethering molecules with an affinity to a bacterial cell under investigation. ### Claim 3 (depends on 1) The system of claim 1 wherein the tethering molecules comprise antibodies. ### Claim 4 (depends on 1) The system of claim 1 wherein the analyte comprises a bacteria selected from the group consisting of E. coli and S. aureus. ### Claim 5 (depends on 1) The system of claim 1 wherein the tethering molecules comprise anti-E. coli antibodies. ### Claim 6 (depends on 1) The system of claim 1 further comprising adhesion materials applied to the metallic coated surface, where the adhesion materials are selected from the group consisting of cell-adhesion promoting substances, poly-lysine, and agar matrix. ### Claim 7 (depends on 1) The system of claim 1 wherein the computer program for tracking 3D motion values tracking the XY-motion comprises a curve fitting algorithm. ### Claim 8 (depends on 7) The system of claim 7 wherein the curve-fitting algorithm is selected from the group consisting of Gaussian fitting, elliptical fitting, and spatial averaging. ### Claim 9 (depends on 8) The system of claim 8 wherein the computer program further comprises a subprogram for extracting an image intensity change from the plasmonic image that is free of noise by transforming the plasmonic image into K-space using Fourier transforms to produce a two-ring image. ### Claim 10 A system for antibiotic susceptibility testing using plasmonic imaging for bacterial cells comprising: a computer processor; a plasmonic imaging and tracking (PIT) system coupled to the computer processor, the PIT including an inverted microscope lens, a light source located to impinge on the inverted microscope lens, a slide, located to receive light from the microscope lens, the slide having a metallic coated surface, a mirror, located to receive reflected light from the metallic coated surface through the inverted microscope lens and transmit the reflected light, and a detector located to receive the reflected light from the mirror; where the metallic coated surface has antibodies attached as tethering molecules; bacteria populating the metallic coated surface; wherein the bacteria is selected from the group consisting of E. coli and S. aureus; a device for adding an antibiotic to the metallic coated surface after it has been populated with bacteria; where the computer processor includes and is programmed to execute a computer program for tracking a first set of 3D motion changes of the bacteria before addition of the antibiotic, then tracking a second set of 3D motion values of the bacteria in the presence of the antibiotic; and then comparing the first and second 3D motion values to determine changes in the 3D motion of the bacteria after addition of the antibiotic. ### Claim 11 (depends on 10) The system of claim 10 wherein the tethering molecules comprise anti-E. coli antibodies. ### Claim 12 (depends on 10) The system of claim 10 further comprising adhesion materials applied to the metallic coated surface, where the adhesion materials are selected from the group consisting of cell-adhesion promoting substances, poly-lysine, and agar matrix. ### Claim 13 (depends on 10) The system of claim 10 wherein the computer program for tracking 3D motion values tracking the XY-motion comprises a curve fitting algorithm. ### Claim 14 (depends on 13) The system of claim 13 wherein the curve-fitting algorithm is selected from the group consisting of Gaussian fitting, elliptical fitting, and spatial averaging. ### Claim 15 (depends on 14) The system of claim 14 wherein the computer program further comprises a subprogram for extracting an image intensity change from the plasmonic image that is free of noise by transforming the plasmonic image into K-space using Fourier transforms to produce a two-ring image. ### Claim 16 A system for antibiotic susceptibility testing using plasmonic imaging for bacterial cells comprising: a computer processor; a plasmonic imaging and tracking (PIT) system coupled to the computer processor, the PIT including an inverted microscope lens, a light source located to impinge on the inverted microscope lens, a slide, located to receive light from the microscope lens, the slide having a metallic coated surface, a mirror, located to receive reflected light from the metallic coated surface through the inverted microscope lens and transmit the reflected light, and a detector located to receive the reflected light from the mirror; where the metallic coated surface has anti-E. coli antibodies attached as tethering molecules and further has adhesion materials applied to the metallic coated surface, where the adhesion materials are selected from the group consisting of cell-adhesion promoting substances, poly-lysine, and agar matrix; E. coli bacteria populating the metallic coated surface; a device for adding an antibiotic to the metallic coated surface after it has been populated with the E. coli bacteria; where the computer processor includes and is programmed to execute a computer program for tracking a first set of 3D motion changes of the bacteria before addition of the antibiotic, then tracking a second set of 3D motion values of the E. coli bacteria in the presence of the antibiotic; and then comparing the first and second 3D motion values to determine changes in the 3D motion of the E. coli bacteria after addition of the antibiotic. ### Claim 17 (depends on 16) The system of claim 16 wherein the computer program for tracking 3D motion values tracking the XY-motion comprises a curve fitting algorithm. ### Claim 18 (depends on 17) The system of claim 17 wherein the curve-fitting algorithm is selected from the group consisting of Gaussian fitting, elliptical fitting, and spatial averaging. ### Claim 19 (depends on 18) The system of claim 18 wherein the computer program further comprises a subprogram for extracting an image intensity change from the plasmonic image that is free of noise by transforming the plasmonic image into K-space using Fourier transforms to produce a two-ring image.