Notice of Pre-AIA or AIA Status The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA. Status of the Claims Claims 1-20 are pending in the application. Claims 13-20 are withdrawn. Claims 1-12 are the subject of this office action. Election/Restrictions Applicant’s election of Group I, claims 1-12 in the reply filed on 8 December 2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)). Priority The instant application claims provisional application 63/265,063, filed 7 December 2021. Claim Rejections - 35 USC § 112(b) The following is a quotation of 35 U.S.C. 112(b): (b ) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention. The following is a quotation of 35 U.S.C. 112 (pre-AIA), second paragraph: The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the appl icant regards as his invention. Claim s 7 , 10 and 11 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. Claim 7 is vague regarding “the method step”. There is no prior introduction of “a method step”, therefore there is insufficient antecedent basis for this limitation in the claims. A broad range or limitation together with a narrow range or limitation that falls within the broad range or limitation (in the same claim) may be considered indefinite if the resulting claim does not clearly set forth the metes and bounds of the patent protection desired. See MPEP § 2173.05(c). In the present instance, claim 10 recites the broad recitation cytokines, and the claim also recites particular species of cytokines which is the narrower statement of the range/limitation. The claim(s) are considered indefinite because there is a question or doubt as to whether the feature introduced by such narrower language is (a) merely exemplary of the remainder of the claim, and therefore not required, or (b) a required feature of the claims. A broad range or limitation together with a narrow range or limitation that falls within the broad range or limitation (in the same claim) may be considered indefinite if the resulting claim does not clearly set forth the metes and bounds of the patent protection desired. See MPEP § 2173.05(c). In the present instance, claim 11 recites the broad recitation microscopy, and the claim also recites specific species of microscopy (i.e. SEM and TEM) which is the narrower statement of the range/limitation. The claim(s) are considered indefinite because there is a question or doubt as to whether the feature introduced by such narrower language is (a) merely exemplary of the remainder of the claim, and therefore not required, or (b) a required feature of the claims. Claim Rejections - 35 USC § 103 The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action: A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made. Claim s 1, 3-6, 8, and 10-12 are rejected under 35 U.S.C. 103 as being unpatentable over Zhu et al (Detecting cytokine release from single T-cells. Anal Chem. 2009 Oct 1;81(19):8150-6.) in view of Singamaneni et al (WO 2020/072924 A1) . Regarding claim s 1 , 3-4, 6, 8, and 10 - 12 Zhu teaches a method for detection cellular protein secretion (Abstract) comprising: Adding at least one stimulant to a sample comprising at least one cell fixed on a substrate, wherein the at least one stimulant stimulates the at least one cell to secrete at least one protein ( Pg. 8152, Col. 1, Par. 2: a mixture of purified anti-CD4 and anti- IFNg amma antibodies was dispensed onto a substrate to create antibody spots; Fig. 1: wherein the substrate comprises microwells on a PEGylated glass substrate; Pg. 8153, Col. 1, Par. 1-4: RBC-lysed blood sample was added into the microfluidic device and allowed to bind to the capture antibodies immobilized on the substrate (anti-CD4). To commence cytokine production, cells were stimulated with PMA and ionomycin) ; Capturing the at least one protein secreted by the at least one cell ( Pg. 8152, Col. 1, Par. 2: a mixture of purified anti-CD4 and anti- IFNg amma antibodies was dispensed onto a substrate to create antibody spots ); Adding at least one detection antibody to the sample ( Pg. 8153, Col. 1, Par. 1-4: at the end of the desired stimulation period, T cells captured in the microwells were exposed to reagents for the detection of IFNg amma production. After washing away the mitogenic solution, microfluidic chambers were filled with biotinylated anti- IFNg amma antibodies for 1h at room temperature ); Adding a fluor to the sample ( Pg. 8153, Col. 1, Par. 4: after addition of detection antibody, chambers were again flushed and filled with streptavidin-Alexa546 for 30 min in order to detect Ab-cytokine complex ); and Measuring at least one fluorescence signal from the sample ( Pg. 8153, Col. 1, Par. 4: The fluorescently labeled cytokines and cells were then visualized and imaged with a confocal microscope; Figs. 2-3 ) (i.e. a plurality of fluorescence signals are measured from a plurality of fluorescent dots ; and wherein the recited microscopy imaging is understood to read on measuring at least one spatial signal from the sample, wherein the at least one spatial signal is measured with a technique selected from the group consisting of microscopy, SEM, TEM, and combinations thereof). Zhu differs from the instant claim in that it does not specifically teach that the fluor added to the sample is a pla s monic-fluor. Regarding claims 1, 3-4, 6, 8, 10, and 12 , Singamaneni discloses a fluorescent nanoconstruct which comprises a plasmonic nanostructure and a fluorescent agent (i.e. plasmonic-fluor) (Abstract). Singamaneni teaches that the plasmonic fluor can be used as an enhanced label in a number of different fluoroimmunoassay formats and applications, and teaches that it is advantageous over traditional fluorophore labels . Singamaneni teaches specifically that the plasmonic fluor can be used in an application for the detection of a target analyte which comprises binding the target analyte with a biotinylated antibody and subsequently adding a streptavidin-conjugated fluorescent label (as in the method of Zhu) (Par. 140: the plasmonic-fluor is useful for enhancing the bioanalytical parameters (sensitivity, LOD, and dynamic range) of fluoroimmunoassays of a number of different formats; Par. 191: the plasmonic-fluor can be used as a reporter molecule in an immunoassay. Primary detection antibody is used to detect a target analyte, and plasmon-fluor is used to report on the concentration of detection antibody present, which is proportional to the amount of target analyte. This can be don by binding a streptavidin conjugate plasmonic-fluor to a biotinylated detection antibody which is already bound to the target analyte). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of Zhu to specifically comprise using a streptavidin-conjugated plasmonic-fluor label, as taught by Singamaneni . One of ordinary skill in the art would be motivated to make this modification because Singamaneni teaches that the disclosed plasmonic-fluor is an ultrabright label which can be used to improve the bioanalytical parameters of fluoroimmunoassays such as the one disclosed by Zhu ( Singamaneni , Par. 3, 140, 191). One of ordinary skill in the art would have a reasonable expectation of success in making this modification because Singamaneni specifically teaches that the plasmonic-fluor label can be applied to immunoassay formats such as the one taught by Zhu ( Singamaneni . Par. 191). Regarding claim 5 , Zhu differs from the instant claim in that it does not specifically teach the method wherein the fluorescent label used is selected from the group consisting of Cy3, Cy5, and combinations thereof. Regarding claim 5 , Singamaneni teaches that the plasmonic-fluor may be tuned to have LSPR wavelengths matching the excitation maximum wavelength of the fluorophore, wherein tuning and wavelength matching can significantly increase the fluorescence intensity of the label (Par. 154-157). Singamaneni teaches that Cy3 and Cy5 are exemplary fluorophores which can be used in these plasmonic- fluors (Par. 157, 169). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have further modified the method of Zhu in view of Singamaneni such that the plasmonic-fluor specifically comprises a fluorescent label selected from the group consisting of Cy3, Cy5, and combinations thereof, as taught by Singamaneni . One of ordinary skill in the art would be motivated to make this modification because Singamaneni teaches that specific interaction and wavelength matching between the plasmonic element and the fluorescent label of the plasmonic fluor can impact the fluorescence intensity of the label, and teaches that Cy3 and Cy5 are exemplary fluorophores which can be used in the plasmonic- fluors . As such, one would be motivated to use the appropriate fluorescent label (such as Cy3 or Cy5) for the chosen application and for the chosen plasmonic fluor and plasmonic nanostructure , in order to achieve a desired fluorescence intensity. One of ordinary skill in the art would have a reasonable expectation of success in making this modification because both Zhu and Singamaneni disclose fluorescence immunoassays which use streptavidin-conjugated fluorescence labels. Claim s 1-8 and 10-12 are rejected under 35 U.S.C. 103 as being unpatentable over Shirasaki et al (Real-time single-cell imaging of protein secretion. Sci Rep. 2014 Apr 22) in view of Singamaneni et al (WO 2020/072924 A1 ) . Regarding claims 1-4, 6-8, and 10 - 12 , Shirasaki teaches a method for detecting cellular protein secretion (Abstract), comprising: Adding at least one stimulant to a sample comprising at least one cell fixed on a substrate, wherein the at least one stimulant stimulates the at least one cell to secrete at least one protein (Pg. 7, Col. 1-2: substrate comprises PDMS microwells formed on a glass substrate. Capture antibodies for IL-1Beta and IL-6 are immobilized on the substrate. Human peripheral blood monocytes isolated from venous blood sample are introduced to the chip and allowed to adhere. Cells are stimulated with LPS to induce cytokine secretion; Fig. 3); Capturing the at least one protein secreted by the at least one cell (Pg. 7, Col. 1, Par. 3-7: Capture antibodies for IL-1Beta and IL-6 are immobilized on the substrate); Adding at least one detection antibody to the sample and adding at least one fluor to the sample (Pg. 7, Col. 2, Par. 1-3: detection antibody (anti-IL-6) labeled with biotin was coupled with CF-labeled streptavidin to produce a detection media comprising CF-labeled detection antibody. After cells have been added to microwells and stimulated with LPS, medium was replaced with fresh detection medium ); and Measuring at least one fluorescence signal from the sample ( Pg. 7, Col. 2, Par. 1-3: monocytes in the selected position of microwells were monitored at 1 min intervals by multichannel microscopy i.e. DIA images, epifluorescence images for calcein and SYTOX and TIRF images for CF660R. The MFIs of each microwells at each time point were measured with the NIS Elements Imaging software ) (i.e. measuring a plurality of fluorescence signals from a plurality of fluorescent dots ; and wherein the recited microscopy imaging is understood to read on measuring at least one spatial signal from the sample, wherein the at least one spatial signal is measured with a technique selected from the group consisting of microscopy, SEM, TEM, and combinations thereof) . Shirasaki differs from the instant claim in that it does not teach that the fluor added to the sample is a plasmonic fluor. Regarding claims 1-4, 6-8, 10 and 12, , Singamaneni discloses a fluorescent nanoconstruct which comprises a plasmonic nanostructure and a fluorescent agent (i.e. plasmonic-fluor) (Abstract). Singamaneni teaches that the plasmonic fluor can be used as an enhanced label in a number of different fluoroimmunoassay formats and applications, and teaches that it is advantageous over traditional fluorophore labels. Singamaneni teaches specifically that the plasmonic fluor can be used in an application for the detection of a target analyte which comprises binding the target analyte with a biotinylated antibody and subsequently adding a streptavidin-conjugated fluorescent label (Par. 140: the plasmonic-fluor is useful for enhancing the bioanalytical parameters (sensitivity, LOD, and dynamic range) of fluoroimmunoassays of a number of different formats; Par. 191: the plasmonic-fluor can be used as a reporter molecule in an immunoassay. Primary detection antibody is used to detect a target analyte, and plasmon-fluor is used to report on the concentration of detection antibody present, which is proportional to the amount of target analyte. This can be done by binding a streptavidin conjugate plasmonic-fluor to a biotinylated detection antibody which is already bound to the target analyte). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of Shirasaki to specifically comprise using a streptavidin-conjugated plasmonic-fluor label which is added to a sample after a biotinylated antibody, as taught by Singamaneni . One of ordinary skill in the art would be motivated to make this modification because Singamaneni teaches that the disclosed plasmonic-fluor is an ultrabright label which can be used to improve the bioanalytical parameters of fluoroimmunoassays such as the one disclosed by Shirasaki ( Singamaneni , Par. 3, 140, 191), and specifically teaches that use of the label may comprise adding a biotinylated detection antibody to a sample followed by adding a streptavidin-conjugated plasmonic-fluor to the sample. One of ordinary skill in the art would have a reasonable expectation of success in making this modification because Singamaneni specifically teaches that the plasmonic-fluor label can be applied to immunoassay formats such as the one taught by Shirasaki ( Singamaneni . Par. 140, 191). Regarding claim 5 , Shirasaki differs from the instant claim in that it does not specifically teach the method wherein the fluorescent label used is selected from the group consisting of Cy3, Cy5, and combinations thereof. Regarding claim 5 , Singamaneni teaches that the plasmonic-fluor may be tuned to have LSPR wavelengths matching the excitation maximum wavelength of the fluorophore, wherein tuning and wavelength matching can significantly increase the fluorescence intensity of the label (Par. 154-157). Singamaneni teaches that Cy3 and Cy5 are exemplary fluorophores which can be used in these plasmonic- fluors (Par. 157, 169). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have further modified the method of Shirasaki in view of Singamaneni such that the plasmonic-fluor specifically comprises a fluorescent label selected from the group consisting of Cy3, Cy5, and combinations thereof, as taught by Singamaneni . One of ordinary skill in the art would be motivated to make this modification because Singamaneni teaches that specific interaction and wavelength matching between the plasmonic element and the fluorescent label of the plasmonic fluor can impact the fluorescence intensity of the label, and teaches that Cy3 and Cy5 are exemplary fluorophores which can be used in the plasmonic- fluors . As such, one would be motivated to use the appropriate fluorescent label (such as Cy3 or Cy5) for the chosen application and for the chosen plasmonic fluor, in order to achieve a desired fluorescence intensity. One of ordinary skill in the art would have a reasonable expectation of success in making this modification because both Shirasaki and Singamaneni are disclose fluorescence immunoassays which use streptavidin-conjugated fluorescence labels. Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over Zhu et al ( Detecting cytokine release from single T-cells. Anal Chem. 2009 Oct 1;81(19):8150-6. ) in view of Singamaneni et al ( WO 2020/072924 A1 ) , as applied to claim 1 above, and further in view of Lore et al ( Quantitative single cell methods that identify cytokine and chemokine expression in dendritic cells. J Immunol Methods. 2001 Mar 1;249(1-2):207-22. ) ADDITIONALLY Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over Shirasaki et al ( Real-time single-cell imaging of protein secretion. Sci Rep. 2014 Apr 22 ) in view of Singamaneni et al (WO 2020/072924 A1) , as applied to claim 1 above, and further in view of Lore et al ( Quantitative single cell methods that identify cytokine and chemokine expression in dendritic cells. J Immunol Methods. 2001 Mar 1;249(1-2):207-22. ). Both Zhu and Shirasaki disclose methods of measuring cytokine release from stimulated immune cells (Zhu, abstract; Shirasaki , abstract) . Both Zhu and Shirasaki differ from claim 9 in that neither reference teaches that the at least one cell is dendritic cells, JAWS II cells, or combinations thereof. Regarding claim 9, Lore teaches methods of measuring cytokine release from dendritic cells stimulated with LPS (Abstract). Lore teaches that studying cytokine release from dendritic cells is useful for understanding the mechanics and patterns of cytokine release from immune cells which can provide insight into immune system regulation and function (Pg. 217, Col. 2, last Par.; Pg. 208, Col. 1, Par. 1-3). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of Zhu in view of Singamaneni and/or the method of Shirasaki in view of Singamaneni to specifically comprise dendritic cells as the at least one cell in the sample for the purpose of measuring cytokine release from dendritic cells, as taught by Lore. One would be motivated to make this modification because Lore teaches that detecting cytokine release from dendritic cells is useful for understanding the mechanics and patterns of cytokine release from immune cells and for providing insight into immune system function and regulation. One of ordinary skill in the art would have a reasonable expectation of success in making this modification because Zhu and Lore and Shirasaki and Lore are all directed to methods of measuring cytokine release from stimulated immune cells. Conclusion The prior art made of record and not relied upon is considered pertinent to applicant's disclosure. Luan et al (Ultrabright fluorescent nanoscale labels for the femtomolar detection of analytes with standard bioassays. Nat Biomed Eng. 2020 May;4(5):518-530. Epub 2020 Apr 20.): discloses applications of plasmonic- fluors in immunoassays Any inquiry concerning this communication or earlier communications from the examiner should be directed to FILLIN "Examiner name" \* MERGEFORMAT ELLIS LUSI whose telephone number is FILLIN "Phone number" \* MERGEFORMAT (571)270-0694 . The examiner can normally be reached FILLIN "Work Schedule?" \* MERGEFORMAT M-Th 8am-6pm ET . Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice. 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If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /ELLIS FOLLETT LUSI/ Examiner, Art Unit 1677 /CHRISTOPHER L CHIN/ Primary Examiner, Art Unit 1677