# US 11,079,388 B2 · Super-resolution immunofluorescence with diffraction-limited preview > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,079,388 B2 - Document ID: US11079388B2 - Status: Granted - Kind: B2 - Application number: 16/094,954 - Filed: 2017-04-25 - Published: 2019-04-25 - Granted: 2021-08-03 - Earliest claimed: 2016-04-26 - Assignees: Ultivue, Inc. - Applicants: Ultivue, Inc. - Inventors: Xi Chen - Web page: https://patentgrep.com/patent/US11079388B2 ## CPC - G01N 33/582: {with fluorescent label} - C07K 16/18: against material from animals or humans - G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)} - G01N 33/533: with fluorescent label - 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) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract An imaging agent comprises (a) at least one target recognition moiety; (b) at least one observable moiety non-transiently bound to the target recognition moiety, and (c) at least one docking moiety bound to the target recognition moiety, wherein the docking moiety is capable of transiently binding at least one observable moiety. In some embodiments, the at least one target recognition moiety is an antibody or antigen binding fragment thereof. A method of performing super-resolution imaging on a sample comprises (a) contacting the sample with at least one imaging agent comprising (i) a target-recognizing molecule non-transiently attached to at least one observable moiety and (ii) a target-recognizing molecule attached to at least one docking moiety, wherein the docking moiety is capable of transiently binding at least one observable moiety; (b) imaging the target-recognizing molecule non-transiently bound to at least one observable moiety; and (c) providing at least one observable moiety capable of transiently binding to the docking moiety and imaging the observable moiety transiently bound to the docking moiety. ## Claims ### Claim 1 A method of performing super-resolution imaging on a sample comprising: a. contacting the sample with at least one imaging agent comprising (i) a target recognition moiety non-transiently attached to at least one first observable moiety and (ii) a target recognition moiety attached to at least one docking moiety, wherein the docking moiety is capable of transiently binding at least one second observable moiety; b. imaging the target recognition moiety non-transiently bound to at least one first observable moiety; and c. providing the at least one second observable moiety capable of transiently binding to the docking moiety and imaging the second observable moiety transiently bound to the docking moiety. ### Claim 2 (depends on 1) The method of claim 1, wherein the target recognition moiety is an antibody or antigen binding fragment thereof. ### Claim 3 (depends on 1) The method of claim 1, wherein the imaging agent is an imaging agent comprising: a. at least one target recognition moiety; b. at least one first observable moiety non-transiently bound to the target recognition moiety, and c. at least one docking moiety bound to the target recognition moiety, wherein the docking moiety is capable of transiently binding at least one second observable moiety. ### Claim 4 (depends on 1) The method of claim 1, further comprising removing or inactivating the observable moiety non-transiently attached to the target recognition moiety. ### Claim 5 A method of performing super-resolution imaging comprising: a. providing an imaging agent comprising (a) at least one target recognition moiety; (b) at least one first observable moiety non-transiently bound to the target recognition moiety, and (c) at least one docking moiety bound to the target recognition moiety, wherein the docking moiety is capable of transiently binding at least one second observable moiety, b. obtaining a preview image using the at least one first observable moiety non-transiently bound to the target recognition moiety; c. assessing the preview image; and d. obtaining a super-resolution image using the at least one second observable moiety transiently bound to the docking moiety. ### Claim 6 (depends on 5) The method of claim 5, wherein the at least one target recognition moiety is an antibody or antigen binding fragment thereof. ### Claim 7 (depends on 5) The method of claim 5, wherein the first observable moiety non-transiently bound to the target recognition moiety is a signal-emitting moiety. ### Claim 8 (depends on 7) The method of claim 7, wherein the signal-emitting moiety is a fluorophore. ### Claim 9 (depends on 5) The method of claim 5, wherein the first observable moiety non-transiently bound to the target recognition moiety is directly attached to the target recognition moiety. ### Claim 10 (depends on 5) The method of claim 5, wherein the first observable moiety non-transiently bound to the target recognition moiety is indirectly attached to the target recognition moiety. ### Claim 11 (depends on 10) The method of claim 10, wherein the first observable moiety non-transiently bound to the target recognition moiety is directly attached to the docking moiety. ### Claim 12 (depends on 5) The method of claim 5, wherein the first observable moiety non-transiently bound to the target recognition moiety is bound covalently. ### Claim 13 (depends on 5) The method of claim 5, wherein the docking moiety is attached to the imaging agent covalently. ### Claim 14 (depends on 5) The method of claim 5, wherein the first observable moiety non-transiently bound to the target recognition moiety is bound noncovalently. ### Claim 15 (depends on 5) The method of claim 5, wherein the docking moiety is attached to the imaging agent noncovalently. ### Claim 16 (depends on 14) The method of claim 14, wherein the first observable moiety non-transiently bound to the target recognition moiety is bound through a nucleic acid-nucleic acid interaction, wherein a. the nucleic acid bound to the first observable moiety is longer than the nucleic acid bound to the target recognition moiety, or b. the nucleic acid bound to the target recognition moiety is longer than the nucleic acid bound to the first observable moiety, and the longer nucleic acid serves as a toehold for displacement of the shorter strand. ### Claim 17 (depends on 5) The method of claim 5, wherein the docking moiety comprises nucleic acids. ### Claim 18 (depends on 5) The method of claim 5, wherein the target recognition moiety is an antibody or antigen binding fragment thereof, an aptamer, or an oligonucleotide. ### Claim 19 (depends on 5) The method of claim 5, wherein both the first observable moiety non-transiently bound to the target recognition moiety and the second observable moiety transiently bound to the target recognition moiety are attached to a single target recognition moiety. ### Claim 20 (depends on 5) The method of claim 5, wherein the first observable moiety non-transiently bound to the target recognition moiety and the second observable moiety transiently bound to the docking moiety attached to the target recognition moiety are attached to different target recognition moieties, wherein: a. the different target recognition moieties bind to the same epitope on a target; b. the different target recognition moieties bind to different epitopes on the same target; or c. the different target recognition moieties bind to different proteins, optionally wherein the proteins are capable of interacting with each other.