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 .
Response to Amendment
The Amendment filed 12/15/2025 has been entered. Claims 1-11, 13-21 and 23-43 remain pending in the application. Claims 11, 13-20 and 31-40 are withdrawn. New grounds of rejections necessitated by amendments are discussed below.
Claim Objections
Claim 1 is objected to because of the following informalities: In line 10, it is suggested to recite “the plurality of different obtained images” as “the plurality of different images” to improve consistency of terminology. Appropriate correction is required.
Claim 29 is objected to because of the following informalities: In line 9, it is suggested to remove the second period after “said fluid analyzer”, i.e. there is an extra period. Appropriate correction is required.
Claim 43 is objected to because of the following informalities: In line 4, it is suggested to include a period after “camera”. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claim 43 is rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
Regarding claim 43, claim 43 recites new limitation “wherein the particles and aggregations thereof in a first of said plurality of images are different from a second one of said plurality of images due to flow of said particles into and out of a field of view of said camera.” The specification discloses “performing image analysis may include obtaining a plurality of different images and analyzing the dynamics of formation of the one or more first aggregates in each image” (page 4, lines 30 - page 5, line 2; page 6, lines 1-3; page 7, lines 23-24; page 13, lines 9-11; page 49, lines 24-26). However, the disclosure fails to describe: the method as claimed “wherein the particles and aggregations thereof in a first of said plurality of images are different from a second one of said plurality of images due to flow of said particles into and out of a field of view of said camera”. Additionally, applicant has not pointed out where the new claim limitation is supported (Remarks filed 12/15/2025). See MPEP 2163.04 (I). Thus, the claims were not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
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 applicant regards as his invention.
Claims 4 and 43 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.
Regarding claim 4, claim 4 recites the limitation "the one or more aggregates" in line 3. There is insufficient antecedent basis for this limitation in the claim.
Regarding claim 43, claim 43 recites the limitation "the camera" in line 3. There is insufficient antecedent basis for this limitation in the claim. For examination purposes, “camera” is interpreted as the same as “imager” of claim 1. It is suggested to recite “the camera” as “the imager” if referring to the same element of claim 1.
Claim Rejections - 35 USC § 101
35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
Claims 1-10, 21, 23-30 and 41-43 are rejected under 35 U.S.C. 101 because the claimed invention is directed to an abstract idea without significantly more.
Claim 1 recites “determining a concentration of at least one target analyte in the flowing fluid sample based on said image analysis”.
In accordance with MPEP 2106, the claims are found to recite statutory subject matter (Step 1: YES) and are analyzed to determine if the claims recite any concepts that equate to an abstract idea, law of nature or natural phenomenon (Step 2A: Prong 1).
In the instant application, the limitations of “analyze dynamics of aggregation” and “determining a concentration of at least one target analyte in the flowing fluid sample based on said image analysis” covers performance of a limitation in the mind, i.e. mental process or mathematical calculation. Other than “controller circuitry”, if the claim limitations, under its broadest reasonable interpretation, covers performance of the limitations in the mind but for the recitation of generic computer components, then the claim limitations fall within the “Mental Processes” grouping of abstract ideas (MPEP 2106.05(f)). Regarding the limitations of “determining a concentration”, the instant specification, page 28, lines 21-30 discusses an expression for calculating concentration, which could be performed mentally or by math. Accordingly, the claims recite abstract ideas (Step 2A: Prong 1: Yes).
This judicial exception is not integrated into a practical application because the claims do not recite any additional elements that reflects an improvement to technology or applies or uses the judicial exception in some other meaningful way (Step 2A, Prong 2: No). In claim 1, after the steps of “analyze dynamics of aggregation” and “determining a concentration…”, there are no further actions performed that integrates the abstract ideas into a practical application. The claimed limitations do not integrate the abstract idea into a practical application because it does not impose any meaningful limits on practicing the abstract idea. The claim does recite “performing a particle-based immunoassay on a flowing fluid sample that is flowing through a channel; providing a fluid analyzer…imaging…performing image analysis…”, however these steps are data gathering steps, wherein data gathering to be used in the abstract idea is an insignificant extra-solution activity, and not a practical application (see MPEP 2106.05(g)). Additionally, the claim does recite a “controller circuitry”, however, the controller circuitry is recited at a high-level of generality (i.e., as generic application operating on a generic computer) such that it amounts no more than mere instructions to apply the exception using a generic computer component; wherein a general purpose computer is not a particular machine (MPEP 2106.05(b)). Thus, the claims are directed to an abstract idea that is not integrated into a practical application (Step 2A, Prong 2: No).
The claims (1-10, 21, 23-30, and 41-43) do not include additional elements that are sufficient to amount to significantly more than the judicial exception. Claim 1 additionally recites “performing a particle-based immunoassay on a flowing fluid sample that is flowing through a channel; providing a fluid analyzer…imaging…performing image analysis…”, however these steps are data gathering steps, wherein data gathering to be used in the abstract idea is an insignificant extra-solution activity, and not a practical application (see MPEP 2106.05(g)). Additionally, the limitations of “performing a particle-based immunoassay on a flowing fluid sample that is flowing through a channel; providing a fluid analyzer…imaging…performing image analysis…” are well-understood, routine and conventional activities as evidenced by at least Dou et al. (US 20170343466 A1), Gibbons et al. (US 20120309636 A1; cited in the OA filed 07/26/2024), and Cheng et al. (US 20160334396 A1; cited in the IDS filed 06/01/2023). See MPEP 2106.05(d). Further, dependent claims 2-10, 21, 23-30, and 41-43 further recite limitations that further limit the abstract idea of the image analysis, include data gathering steps, and include additional limitations that are well-understood, routine and conventional activities as evidenced by Dou et al. (US 20170343466 A1), Gibbons et al. (US 20120309636 A1; cited in the OA filed 07/26/2024), Putnam et al. (US 20150087559 A1), Cheng et al. (US 20160334396 A1; cited in the IDS filed 06/01/2023), Iversen et al. (US 20170356834 A1), Berliner (US 20020001402 A1), Bransky et al. (US 20160199834 A1) and Oku et al. (US 6106778 A). See MPEP 2106.05(d). The additional elements of the claims and dependent claims (2-10, 21, 23-30,41-43) do not comprise an inventive concept when considered individually or as an ordered combination that transforms the claimed judicial exception into a patent-eligible application of the judicial exception. Therefore, the claims do not amount to significantly more than the judicial exception itself (Step 2B: No). The claims are not patent eligible.
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-2, 4-8, 10, and 41-43 are rejected under 35 U.S.C. 103 as being unpatentable over Dou et al. (US 20170343466 A1) in view of Gibbons et al. (US 20120309636 A1; cited in the OA filed 07/26/2024) and Cheng et al. (US 20160334396 A1; cited in the IDS filed 06/01/2023).
Regarding claim 1, Dou teaches a method of analyzing a fluid sample (abstract, Figs. 1-4, and paragraph [0006] teaches a method for analyzing a sample flowing within a microfluidic chip, i.e. fluid sample), the method comprising:
(a) performing a particle-based immunoassay on a flowing fluid sample that is flowing through a channel (Figs. 1-2 and paragraphs [0006], [0018] teaches mixing assay components of a sample and detection bead/antibody complex to perform an assay on a sample flowing through channels and chambers, the channels and chambers are interpreted as a channel, of a microfluidic device);
(b) providing a fluid analyzer comprising an imager (paragraph [0048], “detection module” that comprises an optical detector, i.e. imager) and a controller circuitry (paragraph [0050], “image analysis program”);
(c) imaging a plurality of different images of said flowing fluid sample over a period of time with the imager, while said flowing fluid sample is continuously flowing through the channel (paragraph [0006] teaches capturing images of the cells flowing within the fluid chip; paragraph [0022] teaches continuous imaging capturing; paragraphs [0018],[0024] teaches detection, i.e. imaging, of the bead/antibody complex flowing through the detection window of the fluidic chip; paragraph [0048] teaches capturing optical images over time);
(d) using the controller circuitry (paragraph [0050], teaches “image analysis program” for analyzing and processing the acquired optical images) for:
(i) performing image analysis of the plurality of different obtained images (paragraph [0050] teaches analyzing and processing the acquired optical images, i.e. plurality of different obtained images) to analyze the particles of said particle-based immunoassay, within the flowing fluid sample by said fluid analyzer (paragraph [0050] teaches analyzing images for cell detection, tracking, and enumeration and also for motion analysis of cells or particles flowing through the detection window), and
(ii) determining a concentration of at least one target analyte in the flowing fluid sample based on said image analysis (paragraph [0073] and Fig. 5 teaches the optical imaging system and software determines the target analyte concentration of the sample, wherein paragraph [0050] teaches the images were captured of the fluid sample flowing through the detection window of the microfluidic chip).
Dou fails to teach: imaging a plurality of different images of said flowing fluid sample over a period of time, with the imager positioned by the controller; performing image analysis of the plurality of different obtained images to analyze dynamics of aggregation of the particles of said particle-based immunoassay, within the flowing fluid sample by said fluid analyzer.
Gibbons teaches systems, devices, and methods for point-of-care and distributed testing services (abstract). Gibbons teaches assays performed on a fluidic device (paragraph [0412]). Gibbons teaches imaging systems (paragraph [0677]), where imaging devices enable the exploration of changes in the sample over time by collecting multiple images and comparing changes in the images over time and space, such as would be evident in an aggregation processes or other changes in the sample over time and space (paragraph [0677]). Gibbons teaches imaging devices may enable more rapid data acquisition of arrays, tissue sections, and other assay/sample configurations (paragraph [0677]). Gibbons teaches an embodiment of identifying, locating, and counting red cells and agglutinates in an agglutination reaction with image recognition software (paragraph [0930]). Gibbons teaches reaction mixtures are incubated and introduces into microchannels and imaged, where multiple images are taken to get adequate statistics to objectively evaluate the agglutination process (paragraph [0940]). Gibbons teaches imaged based auto-focusing, where an image-based algorithm is used to control the position of an objective to achieve auto-focusing, i.e. imaging with an imager positioned by a controller (paragraph [0654]). Gibbons teaches detection methods can include tracking a molecule based upon size (paragraph [0412]). Gibbons teaches immunoassays that include antigen and/or antibody and agglutination (paragraph [0473]). Gibbons teaches assays based on agglutination of latex particles or red blood cells can be measured by image analysis using software to interpret the number and size of the agglutinates, i.e. dynamics of aggregation (paragraph [0474]). Gibbons teaches images of a channel are collected, cells can be recognized by size and shape, and pattern recognition algorithms may be employed to analyze aggregated cells (paragraph [0681]).
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 imaging of Dou to incorporate the teachings of an image-based algorithm for controlling the position of an imager to achieve auto-focusing of Gibbons (paragraph [0654]) to provide: imaging a plurality of different images of said flowing fluid sample over a period of time, with the imager positioned by the controller. Doing so would have a reasonable expectation of successfully improving sharpness and focusing for imaging of the sample as taught by Gibbons (paragraph [0654]).
Modified Dou fails to teach: performing image analysis of the plurality of different obtained images to analyze dynamics of aggregation of the particles of said particle-based immunoassay, within the flowing fluid sample by said fluid analyzer.
Dou teaches assaying multiple antigens, analytes, or other microparticulates from patient samples (paragraph [0003]), tracking and counting cells of various sizes (paragraphs [0028], [0031]), a lens positioned above or below a cartridge (paragraph [0049]), and quantifying an amount of target analyte binding per bead population (paragraph [0073]).
Gibbons teaches systems, devices, and methods for point-of-care and distributed testing services (abstract). Gibbons teaches assays performed on a fluidic device (paragraph [0412]). Gibbons teaches imaging systems (paragraph [0677]), where imaging devices enable the exploration of changes in the sample over time by collecting multiple images and comparing changes in the images over time and space, such as would be evident in an aggregation processes or other changes in the sample over time and space (paragraph [0677]). Gibbons teaches imaging devices may enable more rapid data acquisition of arrays, tissue sections, and other assay/sample configurations (paragraph [0677]). Gibbons teaches an embodiment of identifying, locating, and counting red cells and agglutinates in an agglutination reaction with image recognition software (paragraph [0930]). Gibbons teaches reaction mixtures are incubated and introduces into microchannels and imaged, where multiple images are taken to get adequate statistics to objectively evaluate the agglutination process (paragraph [0940]). Gibbons teaches detection methods can include tracking a molecule based upon size (paragraph [0412]). Gibbons teaches immunoassays that include antigen and/or antibody and agglutination (paragraph [0473]). Gibbons teaches assays based on agglutination of latex particles or red blood cells can be measured by image analysis using software to interpret the number and size of the agglutinates, i.e. dynamics of aggregation (paragraph [0474]). Gibbons teaches images of a channel are collected, cells can be recognized by size and shape, and pattern recognition algorithms may be employed to analyze aggregated cells (paragraph [0681]).
Cheng teaches quantitative detection of biomarkers and/or other compounds in a fluid sample using functionalized microparticle aggregates (abstract). Cheng teaches a method where micron-scale particles are functionalized to specifically interact with the biomarker being measured and added to the sample to form aggregates, the size and number, i.e. dynamics of aggregation, of which are counted to find a volume fraction and/or number fraction of aggregates in the sample; wherein by comparing the measured volume fraction and/or number fraction of aggregates in the sample to a calibration curve, the concentration of that biomarker may be determined even for biomarkers or other target compounds in samples at very low concentrations (abstract; paragraph [0008]). Cheng teaches measuring the concentration of a compound in a fluid by counting the number and size of compound-microparticle aggregates (paragraph [0012]). Cheng teaches quantitative detection of biomarker is needed for many applications, where the number or volume ratio of aggregates to total particles is proportional to the target molecule concentration, and aggregates were recognized and counted through images (paragraph [0097]). Cheng teaches this detection method will be particularly useful for hospitals or laboratories that need rapid clinical detection but lack immediate access to analytical instrument; wherein the number of the aggregates can be directly linked to the antigen concentration, and wherein the software for counting the particles and aggregates could be used to replace manual counting to further reduce the assay time (paragraph [0102]).
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 modified Dou to incorporate Dou’s teachings of imaging to track and count cells and quantifying a target analyte (paragraph [0028],[0031],[0073]), Gibbons’ teachings of collecting multiple images of a sample over time and space to analyze aggregation processes or other changes in a sample over time and space (paragraph [0677]), tracking a molecule based on size (paragraph [0412]), and analyzing the size of agglutinates (paragraphs [0474],[0681]) and Cheng’s teachings of determining concentration of a biomarker based on dynamics of aggregation of aggregates in a fluid sample (paragraphs [0008],[0012],[0097],[0102]) to provide: performing image analysis of the plurality of different obtained images to analyze dynamics of aggregation of the particles of said particle-based immunoassay, within the flowing fluid sample by said fluid analyzer. Doing so would have a reasonable expectation of successfully improving tracking and analysis of the change over space and time of a sample (Gibbons, paragraph [0677]), enabling objective evaluation of formation of aggregates (Gibbons, paragraph [0940]), and improving quantitative detection of target particles even at very low concentrations for rapid clinical detection (Cheng, paragraphs [0008],[0012],[0097],[0102]).
Regarding claim 2, modified Dou fails to explicitly teach: wherein said performing image analysis with said fluid analyzer comprises analyzing the dynamics of aggregation of one or more aggregates in each of the images of the plurality of different images.
Dou teaches tracking and counting cells of various sizes (paragraphs [0028], [0031]).
Gibbons teaches imaging systems (paragraph [0677]), where imaging devices enable the exploration of changes in the sample over time by collecting multiple images and comparing changes in the images over time and space, such as would be evident in an aggregation processes or other changes in the sample over time and space (paragraph [0677]). Gibbons teaches reaction mixtures are incubated and introduces into microchannels and imaged, where multiple images are taken to get adequate statistics to objectively evaluate the agglutination process (paragraph [0940]). Gibbons teaches assays based on agglutination of latex particles or red blood cells can be measured by image analysis using software to interpret the number and size of the agglutinates, i.e. dynamics of aggregation (paragraph [0474]). Gibbons teaches images of a channel are collected, cells can be recognized by size and shape, and pattern recognition algorithms may be employed to analyze aggregated cells (paragraph [0681]).
Cheng teaches a method where micron-scale particles are functionalized to specifically interact with the biomarker being measured and added to the sample to form aggregates, the size and number, i.e. dynamics of aggregation, of which are counted to find a volume fraction and/or number fraction of aggregates in the sample; wherein by comparing the measured volume fraction and/or number fraction of aggregates in the sample to a calibration curve, the concentration of that biomarker may be determined even for biomarkers or other target compounds in samples at very low concentrations (abstract; paragraph [0008]). Cheng teaches measuring the concentration of a compound in a fluid by counting the number and size of compound-microparticle aggregates (paragraph [0012]). Cheng teaches quantitative detection of biomarker is needed for many applications, where the number or volume ratio of aggregates to total particles is proportional to the target molecule concentration, and aggregates were recognized and counted through images (paragraph [0097]). Cheng teaches this detection method will be particularly useful for hospitals or laboratories that need rapid clinical detection but lack immediate access to analytical instrument; wherein the number of the aggregates can be directly linked to the antigen concentration, and wherein the software for counting the particles and aggregates could be used to replace manual counting to further reduce the assay time (paragraph [0102]).
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 image analysis of Dou to incorporate Dou’s teachings of tracking cells (paragraph [0028],[0031]), Gibbons’ teachings of collecting multiple images of a sample over time and space to analyze aggregation processes or other changes in a sample over time and space (paragraph [0677]), tracking a molecule based on size (paragraph [0412]), and analyzing the size of agglutinates (paragraphs [0474],[0681]) and Cheng’s teachings of determining concentration of a biomarker based on dynamics of aggregation of aggregates in a fluid sample (paragraphs [0008],[0012],[0097],[0102]) to provide: wherein said performing image analysis with said fluid analyzer comprises analyzing the dynamics of aggregation of one or more aggregates in each of the images of the plurality of different images. Doing so would have a reasonable expectation of successfully improving tracking and analysis of the change over space and time of a sample as taught by Gibbons (paragraph [0677]), and enable objective evaluation of formation of aggregates as discussed by Gibbons (paragraph [0940]) and improving quantitative detection of target particles even at very low concentrations for rapid clinical detection (Cheng, paragraphs [0008],[0012],[0097],[0102]).
Regarding claim 4, modified Dou further teaches: wherein the dynamics of aggregation comprises at least one selected from the group consisting of rate of formation of the one or more aggregates, size of the one or more aggregates, and a combination thereof (see above claim 1; Dou in view of Gibbons and Cheng includes analyzing dynamics of aggregation, i.e. size of one or more aggregates; Gibbons, paragraph [0474], teaches assays based on agglutination of latex particles or red blood cells can be measured by image analysis using software to interpret the number and size of the agglutinates, i.e. dynamics of aggregation; Cheng, paragraph [0008] teaches measuring size and number of aggregates).
Regarding claim 5, modified Dou further teaches wherein said particles of said particle-based immunoassay comprise a first plurality of particles (Fig. 1 and paragraphs [0036] teaches antibodies; paragraph [0071] teaches multiple population of beads each containing unique capture probes) and, wherein each particle of the first plurality of particles comprises a first antibody that is specific to a first target analyte in said fluid sample (Fig. 1, paragraphs [0036],[0071] teaches a capture probe comprising an antibody, and the capture probe is specific to a target analyte), wherein performing said particle-based immunoassay on the flowing fluid sample that is flowing through a channel further comprises:
providing the first plurality of particles, wherein the first plurality of particles and the first target analyte will bind each other, via the first antibody, to form one or more first aggregates (Figs. 1 and 2D; paragraphs [0071]-[0072] teaches capture probes comprising antibody is provided, and a target analyte binds to the capture probe to form a complex, i.e. aggregate).
Modified Dou fails to explicitly teach: said particles of said particle-based immunoassay comprise a second plurality of particles; wherein each particle of the second plurality of particles comprises a second antibody that is specific to a second target analyte in said fluid sample, wherein the second plurality of particles has different optical property values than said first plurality of particles; wherein performing said particle-based immunoassay on the flowing fluid sample that is flowing through a channel further comprises: providing the second plurality of particles, wherein the second plurality of particles and the second target analyte will bind each other, via the second antibody, to form one or more second aggregates.
Dou teaches embodiments of multiplex detection bead sandwich assay, wherein mixture of beads are impregnated with different fluorescent or other light-based signals that allow differentiation of different populations of beads (paragraphs [0008],[0071]). Dou teaches the sample is exposed to multiple populations of beads, each containing unique capture probes directed to specific antigens or analytes, during the same assay, increasing the cost, efficiency, portability, and usability for point-of-care testing (paragraph [0071]). Dou teaches: methods for assaying at least two antigens, analytes, or other microparticulates, comprising: providing beads comprising a mixture of beads impregnated with different fluorescent or other light-based signals that allow differentiation of different populations of beads, a first detector molecule, a second detector module, and capturing images of cells within the fluidic chip (paragraph [0006]).
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 said particles of said particle-based immunoassay and performing said particle-based immunoassay of modified Dou to incorporate the teachings of multiplex detection of a mixture of capture probes and detection of at least two antigens of Dou (paragraphs [0006],[0008],[0071]) to provide: said particles of said particle-based immunoassay comprise a second plurality of particles; wherein each particle of the second plurality of particles comprises a second antibody that is specific to a second target analyte in said fluid sample, wherein the second plurality of particles has different optical property values than said first plurality of particles; wherein performing said particle-based immunoassay on the flowing fluid sample that is flowing through a channel further comprises: providing the second plurality of particles, wherein the second plurality of particles and the second target analyte will bind each other, via the second antibody, to form one or more second aggregates. Doing so would have a reasonable expectation of successfully improving efficiency, portability, and usability for point-of-care testing of multiple antigens or analytes in a sample as taught by Dou (paragraph [0071]).
Regarding claim 6, Dou further teaches wherein performing image analysis of the flowing fluid sample further comprises:
imaging a plurality of different images of said flowing fluid sample to capture dynamics of formation of the one or more first aggregates (see above claim 1; Dou in view of Gibbons teaches imaging a plurality of different images of said flowing fluid sample with the imager and performing image analysis of the obtained images to analyze dynamics of aggregation of particles of said particle-based immunoassay, thus capturing dynamics of aggregation of formation, i.e. size of one or more aggregates); and
performing image analysis of the obtained images to analyze the dynamics of formation of the one or more first aggregates (see above claim 1; Dou in view of Gibbons teaches performing image analysis of the obtained images to analyze dynamics of aggregation of particles of said particle-based immunoassay, i.e. size of one or more aggregates).
Modified Dou fails to explicitly teach: wherein performing image analysis of the flowing fluid sample further comprises: imaging a plurality of different images of said flowing fluid sample to capture dynamics of formation of the one or more second aggregates; performing image analysis of the obtained images to analyze the dynamics of formation of the one or more first aggregates and the one or more second aggregates to determine the concentration of the first target analyte in the flowing fluid sample and the concentration of the second target analyte in the flowing fluid sample.
Dou teaches embodiments of multiplex detection bead sandwich assay (paragraphs [0008],[0071]). Dou teaches the sample is exposed to multiple populations of beads, each containing unique capture probes directed to specific antigens or analytes, during the same assay, increasing the cost, efficiency, portability, and usability for point-of-care testing (paragraph [0071]). Dou teaches: methods for assaying at least two antigens, analytes, or other microparticulates, comprising: providing beads comprising a mixture of beads impregnated with different fluorescent or other light-based signals that allow differentiation of different populations of beads, a first detector molecule, a second detector module, and capturing images of cells within the fluidic chip (paragraph [0006]). Dou teaches determining a concentration of at least one target analyte in the flowing fluid sample (paragraph [0073] and Fig. 5 teaches the optical imaging system and software determines the target analyte concentration of the sample, wherein paragraph [0050] teaches the images were captured of the fluid sample flowing through the detection window of the microfluidic chip).
Cheng teaches quantitative detection of biomarkers and/or other compounds in a fluid sample using functionalized microparticle aggregates (abstract). Cheng teaches a method where micron-scale particles are functionalized to specifically interact with the biomarker being measured and added to the sample to form aggregates, the size and number of which are counted to find a volume fraction and/or number fraction of aggregates in the sample; wherein by comparing the measured volume fraction and/or number fraction of aggregates in the sample to a calibration curve, the concentration of that biomarker may be determined even for biomarkers or other target compounds in samples at very low concentrations (abstract; paragraph [0008]).
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 performing image analysis of modified Dou to incorporate the teachings of the teachings of multiplex detection of a mixture of capture probes and detection of at least two antigens of Dou (paragraphs [0006],[0008],[0071]) and the teachings of analyzing size of aggregates, i.e. dynamics of formation of aggregates, to determine concentration of target compounds of Cheng (abstract; paragraph [0008]) to provide: wherein performing image analysis of the flowing fluid sample further comprises: imaging a plurality of different images of said flowing fluid sample to capture dynamics of formation of the one or more second aggregates; performing image analysis of the obtained images to analyze the dynamics of formation of the one or more first aggregates and the one or more second aggregates to determine the concentration of the first target analyte in the flowing fluid sample and the concentration of the second target analyte in the flowing fluid sample. Doing so would have a reasonable expectation of successfully improving efficiency, portability, and usability for point-of-care testing of multiple antigens or analytes in a sample as taught by Dou (paragraph [0071]), and further improving calculation of concentration of target analytes at low concentrations (Cheng, abstract).
Regarding claim 7, Dou further teaches wherein the flowing fluid sample comprises intact cells (paragraphs [0006],[0045], teaches cells flowing within the fluidic chip, which is interpreted as intact cells; paragraph [0027]; paragraph [0029] teaches a sample can include whole blood, which comprises intact cells) and the method is conducted in the presence of the intact cells (paragraphs [0006],[0045], teaches capturing images of cells flowing within the fluidic chip; paragraph [0029] teaches a sample to be analyzed can include whole blood, which comprises intact cells).
Regarding claim 8, while Dou teaches detecting capture probes and complexes (paragraph [0071]-[0072]), analysis can include a sample comprising a combination of components, such as cells, antigens, or other microparticulates (paragraph [0027]), and a sample can comprise whole blood (paragraph [0029]), modified Dou fails to teach: wherein the image analysis excludes the intact cells that are present in the imaged fluid sample.
Gibbons teaches pattern recognition algorithms may be employed to exclude stained cell debris and in most cases where there are cells which are aggregated these can either be excluded from the analysis or interpreted as aggregates (paragraph [0681]).
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 image analysis of modified Dou to incorporate the teachings of a pattern recognition algorithm to exclude elements, such as cell debris, and to interpret cell aggregates of Gibbons (paragraph [0681]) and the teachings of detecting capture probes and complexes of Dou (paragraph [0071]-[0072]) and the use of whole blood samples of Dou (paragraph [0029]) to provide: wherein the image analysis excludes the intact cells that are present in the imaged fluid sample. Doing so would have a reasonable expectation of successfully improving detection and analysis of aggregation of particles in a sample that comprises additional components, such as cells in a blood sample.
Regarding claim 10, Dou further teaches the method of claim 1, further comprising:
providing a cartridge comprising the channel (Figs. 2A-2D and paragraph [0006] teaches a microfluidic device comprising channels and chambers, the channels and chambers are interpreted as the channel);
introducing the flowing fluid sample comprising a first target analyte of said at least one target analyte into a reservoir of the cartridge (Figs. 2A-2C and paragraphs [0006],[0071]-[0072] teaches introducing a sample to a microfluidic chamber, wherein the sample comprises a target analyte), the reservoir comprising a first reagent (Figs. 2A-2C and paragraph [0036] teaches the microfluidic chamber comprises reagent; Figs. 2A-2B shows the left microfluidic chamber comprises analyte detection beads);
flowing the flowing fluid sample into a second reservoir of the cartridge comprising a first particle of said particle-based immunoassay (paragraph [0072] and Fig. 2C teaches a sample flowing to a second microfluidic chamber comprising analyte detection antibody, i.e. first particle), wherein each particle of the first plurality of particles comprises a first antibody that is specific to the first target analyte (Fig. 2C, “analyte detection antibody”; paragraph [0071] teaches a capture probe binds to a target analyte, therefore the antibody is specific to a first target analyte) and the first particle and the first target analyte will bind each other, via the first antibody, to form one or more first aggregates (Figs. 1 and 2D and paragraphs [0071]-[0072] teaches a capture probe binds to a target analyte, and a detection bead/analyte complex is formed; therefore, the analyte and target analyte will bind via the antibody to form at least one aggregate);
flowing the flowing fluid sample and first particle through said channel in the cartridge (paragraph [0072] and Fig. 2D teaches a detection bead/antibody complex flowing through a detection window);
imaging the flowing fluid sample within said channel in the cartridge to capture dynamics of formation of the one or more first aggregates (paragraph [0006] teaches capturing images of the cells flowing within the fluid chip; paragraph [0022] teaches continuous imaging capturing; paragraphs [0018],[0024] teaches detection, i.e. imaging, of the bead/antibody complex flowing through the detection window of the fluidic chip; therefore, capturing the image would capture the size of the complex, i.e. dynamics of formation of the one or more first aggregate); and
performing the image analysis to determine the concentration of the first target analyte in the flowing fluid sample (paragraph [0073] and Fig. 5 teaches the optical imaging system and software determines the target analyte concentration of the sample, wherein paragraph [0050] teaches the images were captured of the fluid sample flowing through the detection window of the microfluidic chip).
Dou fails to explicitly teach: incubating the flowing fluid sample with the first reagent; flowing the flowing fluid sample into a second reservoir of the cartridge comprising a first plurality of particles of said particle-based immunoassay, wherein each particle of the first plurality of particles comprises a first antibody that is specific to the first target analyte and the first plurality of particles and the first target analyte will bind each other, via the first antibody, to form one or more first aggregates; flowing the flowing fluid sample and first plurality of particles through said channel in the cartridge.
Dou teaches assaying multiple antigens, analytes, or other microparticulates from patient samples (paragraph [0003]), tracking and counting cells of various sizes (paragraphs [0028], [0031]), a lens positioned above or below a cartridge (paragraph [0049]), and quantifying an amount of target analyte binding per bead population (paragraph [0073]). Dou teaches embodiments of chambers with a plurality of antibodies (paragraph [0036]).
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 modified Dou to incorporate the teachings of assaying multiple antigens and analytes and utilizing a plurality of antibodies in a chamber of Dou (paragraphs [0003],[0036]) to provide: flowing the flowing fluid sample into a second reservoir of the cartridge comprising a first plurality of particles of said particle-based immunoassay, wherein each particle of the first plurality of particles comprises a first antibody that is specific to the first target analyte and the first plurality of particles and the first target analyte will bind each other, via the first antibody, to form one or more first aggregates; flowing the flowing fluid sample and first plurality of particles through said channel in the cartridge. Doing so would have a reasonable expectation of successfully improving capturing of the target analyte and thus analysis of the target analyte.
While Dou teaches methods for assaying including flowing a sample through a fluidic chip (paragraph [0006]; Figs. 2A-2D), modified Dou fails to teach: incubating the flowing fluid sample with the first reagent.
Gibbons teaches duration of assay detection can be adjusted accordingly to the type of assay that is to be carried out with a device of the invention; for example, if needed for higher sensitivity, an assay can be incubated for more than one hour or up to more than one day (paragraph [0205]). Gibbons teaches a reagent may be added to a sample, and one or more incubation steps may occur (paragraph [0356]). Gibbons teaches heat can be used in the incubation step of an assay reaction to promote the reaction and shorten the duration necessary for the incubation step (paragraph [0395]).
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 modified Dou to incorporate the teachings of incubating reagents with samples of Gibbons (paragraphs [0205],[0356],[0395]) and the teachings of assaying a sample flowing through a fluidic chip of Dou (paragraph [0006]; Figs. 2A-2D) to provide: incubating the flowing fluid sample with the first reagent. Doing so would have a reasonable expectation of successfully promoting reaction and improving sensitivity of an assay as taught by Gibbons (paragraphs [0205],[0356],[0395]).
Regarding claim 41, Dou further teaches wherein: said imaging comprises obtaining a plurality of different images (paragraph [0006] teaches capturing one or more images; paragraph [0073] teaches images are captured), wherein each image is captured at a different given respective time period (paragraph [0048] teaches capturing optical images over time), and wherein the said performing image analysis comprises analysis of the dynamics of aggregation of the particles within the flowing fluid sample over the given period of time (paragraph [0050] teaches images are captured by the optical detector during a specified time period and the acquired optical images are analyzed and processed for particle and cell detection, tracking, and enumeration; see above claim 1, Dou in view of Gibbons and Cheng includes analyzing dynamics of aggregation, i.e. size of one or more aggregates; therefore, the dynamics of aggregation of the particles, i.e. size, are analyzed over the given period of time that the images are captured; Cheng, paragraph [0008], teaches measuring size and number of aggregates).
Regarding claim 42, modified Dou fails to explicitly teach: wherein the performing image analysis image analysis is performed on the obtained images of particles during the formation of aggregates in said particle-based immunoassay.
Gibbons teaches systems, devices, and methods for point-of-care and distributed testing services (abstract). Gibbons teaches cartridges with fluidic systems (paragraph [0189]). Gibbons teaches assays performed on a fluidic device (paragraph [0412]). Gibbons teaches imaging systems (paragraph [0677]), where imaging devices enable the exploration of changes in the sample over time by collecting multiple images and comparing changes in the images over time and space, such as would be evident in an aggregation processes or other changes in the sample over time and space (paragraph [0677]). Gibbons teaches imaging devices may enable more rapid data acquisition of arrays, tissue sections, and other assay/sample configurations (paragraph [0677]). Gibbons teaches an embodiment of identifying, locating, and counting red cells and agglutinates in an agglutination reaction with image recognition software (paragraph [0930]). Gibbons teaches reaction mixtures are incubated and introduces into microchannels and imaged, where multiple images are taken to get adequate statistics to objectively evaluate the agglutination process (paragraph [0940]).
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 image analysis of modified Dou to incorporate the teachings of collecting images over time of a sample to analyze changes in a sample such as for an aggregation process or agglutination process of Gibbons (paragraphs [0677],[0930],[0940]) to provide: wherein the performing image analysis image analysis is performed on the obtained images of particles during the formation of aggregates in said particle-based immunoassay. Doing so would have a reasonable expectation of successfully improving analysis of the change over space and time of a sample as taught by Gibbons (paragraph [0677]), and enable objective evaluation of formation of aggregates as discussed by Gibbons (paragraph [0940]).
Regarding claim 43, modified Dou fails to teach the method of claim 1, wherein the particles and aggregations thereof in a first of said plurality of images are different from a second one of said plurality of images due to flow of said particles into and out of a field of view of said camera.
Dou teaches a sample including a mixture of beads impregnated with different fluorescent that allows differentiation of different population of beads and capturing one or more images of the one or more cells (paragraphs [0006],[0012]). Dou teaches advantages of analyzing many different types of particulates and continuous image capturing capabilities (paragraph [0022]). Dou teaches a detection window to facilitate capturing of the one or more images of one or more detection beads flowing within the detection window (paragraph [0023]). Dou teaches image analysis that uses images captured by the optical detector of cells or particles flowing through the detection window during a specified time period (paragraph [0050]).
Gibbons teaches assays performed on a fluidic device (paragraph [0412]). Gibbons teaches imaging systems (paragraph [0677]), where imaging devices enable the exploration of changes in the sample over time by collecting multiple images and comparing changes in the images over time and space, such as would be evident in an aggregation processes or other changes in the sample over time and space (paragraph [0677]). Gibbons teaches imaging devices may enable more rapid data acquisition of arrays, tissue sections, and other assay/sample configurations (paragraph [0677]). Gibbons teaches an embodiment of identifying, locating, and counting red cells and agglutinates in an agglutination reaction with image recognition software (paragraph [0930]). Gibbons teaches reaction mixtures are incubated and introduces into microchannels and imaged, where multiple images are taken to get adequate statistics to objectively evaluate the agglutination process (paragraph [0940]).
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 modified Dou to incorporate Dou’s teachings of capturing images of different beads and capturing images as the beads flow within the detection window (paragraphs [0006],[0012],[0022],[0050]) and Gibbons’ teachings of collecting multiple images of reaction mixtures that are introduced into microchannels of Gibbons (paragraphs [0677],[0930],[0940]) to provide: the method of claim 1, wherein the particles and aggregations thereof in a first of said plurality of images are different from a second one of said plurality of images due to flow of said particles into and out of a field of view of said camera. Doing so would have a reasonable expectation of successfully improving image analysis of different particles and aggregations thereof in a fluid sample flowing through the detection window, therefore, improving analysis of many different types of particulates via continuous image capturing (Dou, paragraph [0022]).
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Dou in view of Gibbons and Cheng as applied to claim 1 above, and further in view of Putnam et al. (US 20150087559 A1).
Regarding claim 3, modified Dou fails to teach: wherein the imaging of a plurality of different images of said flowing fluid sample comprises obtaining a vertical scan along a height of the channel with the imager positioned by the controller circuitry.
Gibbons teaches controlling the z-position of an objective to achieve auto-focusing (paragraph [0654]).
Putnam teaches a microfluidic device comprising a microfluidic channel (abstract). Putnam teaches steps of an immunoassay protocol conducted within a cassette (paragraph [0297]). Putnam teaches a scanner comprising a camera (paragraph [0686]), wherein the first step in the automatic scan is to determine where the channels are located relative to the stage x, y positions, and to also determine the skew of the chip in the event that the channels are not exactly parallel with the vertical axis (paragraph [0694]), thus allowing for focusing (paragraph [0704]).
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 step of performing image analysis of modified Dou to incorporate the teachings of scanning a vertical axis of channels of Putnam (paragraphs [0694],[0704]) and the teachings of controlling the z-position of an objective to achieve auto-focusing of Gibbons (paragraph [0654]) to provide: wherein the imaging of a plurality of different images of said flowing fluid sample comprises obtaining a vertical scan along a height of the channel with the imager positioned by the controller circuitry. Doing so would have a reasonable expectation of successfully improving location of channels in the event that the channel is not properly aligned, thus improving auto-focusing and imaging of the channel during image analysis.
Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over Dou in view of Gibbons, Cheng, and Iversen as applied to claim 8 above, and further in view Iversen et al. (US 20170356834 A1) and Berliner (US 20020001402 A1).
Regarding claim 9, modified Dou fails to teach: wherein the intact cells are excluded by a technique comprising: processing an image of the intact cells to produce a background threshold; processing an image of the flowing fluid sample comprising the intact cells and one or more aggregates; and normalizing the image of the flowing fluid sample against the background threshold, thereby excluding the image of the intact cells from the image analysis of the flowing fluid sample.
Iversen teaches a method for quantitative detection of particles in fluid (abstract). Iversen teaches background signal and individual groups of cells are excluded from images to improve evaluation of the sensor signal (paragraph [0045]). Iversen teaches an advantage that individual, greatly contaminated parts of the sample carrier window can be excluded in a targeted manner with the detection, and this also includes larger particles, such as air bubbles or particles in a size magnitude (paragraph [0018]). Iversen teaches fading out the background or background noise to simplify and improve the particle detection (paragraph [0004]).
Berliner teaches a system for generating a profile of particulate components of a body fluid sample, wherein a device causes controlled flow of the body fluid sample and the sample is imaged (abstract). Berliner teaches an image of a control blood sample was taken where most of the red blood cells exist in a non-aggregated state, i.e. intact cells (paragraph [0153]). Berliner teaches the image can be analyzed to exclude presence of proteins (paragraph [0153]). Berliner teaches an image acquired from a blood sample taken during the process of aggregation (paragraph [0154]). Berliner teaches comparing control samples with samples taken from individuals suffering from sepsis (paragraph [0160]). Berliner teaches normalizing using a threshold (paragraph [0168] teaches an image is binarized using a threshold), wherein blobs that are greater than and smaller than thresholds are rejected (paragraphs [0171], [0174],[0175]).
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 modified Dou to incorporate the teachings of comparing a control blood sample image of intact cells with an image of a sample with aggregation of Berliner (paragraphs [0153],[0154],[0160]), the teachings of normalizing an image using a threshold to exclude elements of an image of Berliner (paragraphs [0168], [0171],[0174]-[0175]), and the teachings of background signal and individual groups of cells are excluded from images to improve evaluation of the sensor signal of Iversen (paragraphs [0004],[0018], [0045]) to provide: wherein the intact cells are excluded by a technique comprising: processing an image of the intact cells to produce a background threshold; processing an image of the flowing fluid sample comprising the intact cells and one or more aggregates; and normalizing the image of the flowing fluid sample against the background threshold, thereby excluding the image of the intact cells from the image analysis of the flowing fluid sample. Doing so would have a reasonable expectation of successfully improving distinguishing, detecting, and analysis of the aggregates in a sample that comprises additional components, such as cells in a blood sample.
Claims 21, 23-25, and 27-30 are rejected under 35 U.S.C. 103 as being unpatentable over Dou in view of Gibbons and Cheng as applied to claim 1 above, and further in view of Bransky et al. (US 20160199834 A1) and Oku et al. (US 6106778 A).
Regarding claim 21, Dou further teaches the method further comprising
providing a fluidic device (Fig. 2 and paragraphs [0006], [0018] teaches providing a microfluidic device) comprising a second portion for performing the particle based immunoassay on a flowing fluid sample that is flowing through a channel (Figs. 2A-2D and paragraphs [0006],[0071]-[0072] teaches a chambers and channels of the microfluidic device for performing the assay comprising detection beads and antibody on a flowing fluid sample that flows through the channels and chambers of the microfluidic device);
performing the immunoassay on the flowing fluid sample that is flowing through said channel in the second portion of the fluidic device to obtain results of the immunoassay (Figs. 1-2 and paragraphs [0006], [0018] teaches mixing assay components of a sample and detection bead/antibody complex to perform an assay on a sample flowing through channels and chambers, the channels and chambers are interpreted as a channel, of a microfluidic device; paragraphs [0018],[0024] teaches detection of the bead/antibody complex flowing through the detection window of the fluidic chip, i.e. obtaining results of the immunoassay); and
analysis of the results of the immunoassay of the said flowing fluid sample that is flowing through said channel by said fluid analyzer (paragraph [0050] teaches analyzing and processing the acquired optical images and analyzing images for cell detection, tracking, and enumeration and also for motion analysis of cells or particles flowing through the detection window; Fig. 2D and paragraph [0073] teaches the optical imaging system and software determines the target analyte concentration of the sample, wherein paragraph [0050] teaches the images were captured of the fluid sample flowing through the detection window of the microfluidic chip)
While Dou teaches a sample may comprise whole blood (paragraph [0029]), modified Dou fails to teach: the fluidic device comprising a first portion configured for performing a complete blood count assay; performing the complete blood count assay in the first portion of the fluidic device to obtain a hematocrit; wherein the obtained hematocrit is used in the analysis of results of the immunoassay by said fluid analyzer.
Bransky teaches a disposable cartridge having a fluid analysis chip for receiving a fluid to be analyzed (abstract), specifically the cartridge is for preparing a sample fluid that may contain cells for analysis (paragraph [0002]) and the cartridge may be introduced into a reader system to perform optical analysis of fluid flowing through a flow chamber of the cartridge (paragraph [0002]). Bransky teaches the desire for simple point-of-care testing systems for blood tests that use self-contained disposable cartridges or strips (paragraphs [0003]-[0004]). Bransky teaches two parallel preparation units may enable performance of two separate independent procedures, such as one for performing a complete blood count (paragraph [0087]; Fig. 13A), wherein the units comprise a first portion (Fig. 13A, elements 801,802) and a second portion (701,702). Bransky teaches one preparation unit configured for preparing blood cells for analysis (paragraph [0087]). Bransky teaches two analyzing units (Fig. 12; paragraph [0097]), one comprising a microchannel (1003) and another comprising an analyzing reservoir (1101), wherein parallel arranged analyzing units within an analyzing compartment enable performance in parallel of two separate types of analysis of the output fluid (paragraph [0098]), and different analysis using different analyzing modules such as a camera can be performed (paragraph [0098]). Bransky teaches the system may be configured to perform a complete blood count (paragraph [0036]), wherein a complete blood count is interpreted as providing a hematocrit. Bransky teaches the embodiments simplifies the cartridge design, improves manufacturability, and/or enhances reliability and cartridge functions (paragraph [0006]). Bransky teaches a cartridge for preparing a blood sample for optical analysis resulting in obtaining a complete blood count (paragraph [0048]). Bransky teaches the system enables quick obtaining of laboratory results (paragraph [0049]).
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 modified Dou to incorporate the teachings of performing two separate independent preparation and analysis procedures in a cartridge for analyzing blood of Bransky (paragraphs [0087], [0097]-[0098]; Figs. 12-13A) and the teachings of performing a complete blood count of Bransky (paragraphs [0036],[0048]) to provide: the fluidic device comprising a first portion configured for performing a complete blood count assay; performing the complete blood count assay in the first portion of the fluidic device to obtain a hematocrit. Doing so would have a reasonable expectation of successfully improving performance and efficiency of analyzing a blood sample as discussed by Bransky (paragraph [0098]), and enhancing reliability and functions of the overall method as discussed by Bransky (paragraph [0006]), and thus, providing a simplified sample processing procedure and quick obtaining of laboratory results (Bransky, paragraph [0049]).
Modified Dou fails to teach: wherein the obtained hematocrit is used in the analysis of results of the immunoassay by said fluid analyzer.
Oku teaches a compact combination blood cell count and immunoassay apparatus (abstract). Oku teaches a blood cell count/immunoassay apparatus using whole blood comprises an immunoassay section for measuring immunity and a blood cell measuring section for counting the number of blood cells, wherein the same drawn whole blood sample is used in both measuring sections and at the same time, and the results of immunoassay are corrected using a hematocrit value obtained by the measurement of the number of blood cells (column 1, line 62- column 2, line 3). Oku teaches with respect to CRP measurement, a plasma component volume error by the use of whole blood must be correct to accurately determine CRP concentration (column 7, lines 12-22).
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 modified Dou to incorporate the teachings of correcting results of an immunoassay using a hematocrit value of Oku (column 1, line 62- column 2, line 3) to provide: wherein the obtained hematocrit is used in the analysis of results of the immunoassay by said fluid analyzer. Doing so would improve accurate determination of concentration of the target analyte (Oku, column 7, lines 12-22).
Regarding claim 23, Dou further teaches wherein the immunoassay is performed on whole blood comprising intact cells (paragraph [0029] teaches a sample to be analyzed can include whole blood, which is interpreted as comprising intact cells).
Regarding claim 24, Dou further teaches wherein the immunoassay is performed without lysing the intact cells (Figs. 1-2 and paragraphs [0006],[0071]-[0072] teaches a method where beads coupled to a capture probe, i.e. antibody, binds to the target analyte and is flowed through the detection window, therefore, immunoassay is performed without lysing the intact cells; i.e. the steps do not include a lysing step of intact cells).
Regarding claim 25, while Dou teaches detecting capture probes and complexes (paragraph [0071]-[0072]), analysis can include a sample comprising a combination of components, such as cells, antigens, or other microparticulates (paragraph [0027]), and a sample can comprise whole blood (paragraph [0029]), modified Dou fails to teach: wherein the image analysis excludes the intact cells that are present in the imaged fluid sample.
Gibbons teaches pattern recognition algorithms may be employed to exclude stained cell debris and in most cases where there are cells which are aggregated these can either be excluded from the analysis or interpreted as aggregates (paragraph [0681]).
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 image analysis of modified Dou to incorporate the teachings of a pattern recognition algorithm to exclude elements, such as cell debris, and to interpret cell aggregates of Gibbons (paragraph [0681]) and the teachings of detecting capture probes and complexes of Dou (paragraph [0071]-[0072]) and the use of whole blood samples of Dou (paragraph [0029]) to provide: wherein the image analysis excludes the intact cells that are present in the imaged fluid sample. Doing so would have a reasonable expectation of successfully improving detection and analysis of aggregation of particles in a sample that comprises additional components, such as cells in a blood sample.
Regarding claim 27, Dou further teaches wherein the immunoassay is performed on said flowing fluid sample (Figs. 1-2 and paragraphs [0006], [0018] teaches mixing assay components of a sample and detection bead/antibody complex to perform an assay, i.e. immunoassay, on a sample flowing through channels and chambers), and wherein the fluidic device is a cartridge (Fig. 2 and paragraph [0006] teaches a microfluidic chip, i.e. cartridge; paragraphs [0023],[0034] teaches the fluidic chip is a cartridge) that is configured to be operably coupled to an analytical instrument (Fig. 3 teaches the cartridge is configured to be operably coupled and aligned to a detector and light source), wherein the cartridge is pre-loaded with reagents for the immunoassay (paragraphs [0004],[0010],[0036],[0041]) .
Modified Dou fails to teach: wherein the cartridge is pre-loaded with reagents for each of the complete blood count assay and the immunoassay.
Bransky teaches the desire for simple point-of-care testing systems for blood tests that use self-contained disposable cartridges or strips (paragraphs [0003]-[0004]). Bransky teaches two parallel preparation units may enable performance of two separate independent procedures, such as one for performing a complete blood count (paragraph [0087]; Fig. 13A), wherein the units comprise a first portion (Fig. 13A, elements 801,802) and a second portion (701,702). Bransky teaches two analyzing units (Fig. 12; paragraph [0097]), one comprising a microchannel (1003) and another comprising an analyzing reservoir (1101), wherein parallel arranged analyzing units within an analyzing compartment enable performance in parallel of two separate types of analysis of the output fluid (paragraph [0098]), and different analysis using different analyzing modules such as a camera can be performed (paragraph [0098]). Bransky teaches the system may be configured to perform a complete blood count (paragraph [0036]), wherein a complete blood count is interpreted as providing a hematocrit. Bransky teaches the embodiments simplifies the cartridge design, improves manufacturability, and/or enhances reliability and cartridge functions (paragraph [0006]). Bransky teaches a cartridge for preparing a blood sample for optical analysis resulting in obtaining a complete blood count (paragraph [0048]). Bransky teaches the system enables quick obtaining of laboratory results (paragraph [0049]). Bransky teaches a reservoir is pre-loaded with reagents (paragraph [0060]).
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 cartridge modified Dou to incorporate the teachings of performing a complete blood count of Bransky (paragraphs [0036],[0048]) and reservoirs pre-loaded with reagents of Bransky (paragraph [0060]) to provide: wherein the cartridge is pre-loaded with reagents for each of the complete blood count assay and the immunoassay. Doing so would have a reasonable expectation of successfully improving performance and efficiency of analyzing a blood sample as discussed by Bransky (paragraph [0098]), and enhancing reliability and functions of the overall method as discussed by Bransky (paragraph [0006]), and thus, providing a simplified sample processing procedure and quick obtaining of laboratory results (Bransky, paragraph [0049]).
Regarding claim 28, modified Dou fails to teach: wherein the performing image analysis of the obtained images to analyze dynamics of aggregation comprises at least one analysis of an optical property of morphological and/or color characteristic of aggregates formed by said particles, by said fluid analyzer.
Gibbons teaches images can be analyzed for shape recognition for determining concentration of cells and platelets and for observing the state of the sample (paragraph [0614]) and cells can be recognized by their characteristics fluorescence, size, and shape (paragraph [0681]). Gibbons teaches the image processing algorithms utilized in this step may use combinations of image filtering, edge detection, template matching, automatic thresholding, morphological operations and shape analysis of objects (paragraph [0726]). Gibbons teaches a label, such as a colored compound, is coupled to a molecule to be detected (paragraph [0413]).
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 performing image analysis of modified Dou to incorporate the teachings of image processing algorithms to analyze the morphological operations and shape analysis of objects, and the use of colored compounds for detecting a molecule of Gibbons (paragraphs [0413], [0614],[0681],[0726]) to provide: wherein the performing image analysis of the obtained images to analyze dynamics of aggregation comprises at least one analysis of an optical property of morphological and/or color characteristic of aggregates formed by said particles, by said fluid analyzer. Doing so would have a reasonable expectation of successfully improving characterization and analysis of the aggregation and thus allow for determining of a concentration of the aggregation as taught by Gibbons (paragraph [0614]).
Regarding claim 29, while Dou teaches a multiplex detection assay (paragraph [0071]), Dou fails to teach: wherein the performing of the particle-based immunoassay on a flowing fluid sample that is flowing through a channel comprises performing at least two separate particle based immunoassays, wherein each of the separate particle based immunoassays, provides a separate optical property of morphological and/or color characteristic of aggregates formed by said particles, and wherein the performing image analysis of the obtained images comprises analyses of each separate optical property of morphological and/or color characteristic of aggregates formed by said particles in said separate immunoassays, by said fluid analyzer.
Gibbons teaches images can be analyzed for shape recognition for determining concentration of cells and platelets and for observing the state of the sample (paragraph [0614]) and cells can be recognized by their characteristics fluorescence, size, and shape (paragraph [0681]). Gibbons teaches the image processing algorithms utilized in this step may use combinations of image filtering, edge detection, template matching, automatic thresholding, morphological operations and shape analysis of objects (paragraph [0726]). Gibbons teaches a label, such as a colored compound, is coupled to a molecule to be detected (paragraph [0413]). Gibbons teaches several assay elements can be imaged in parallel (paragraph [0603]), and a sample can be distributed in multiple containers for sequential or parallel processing and imaging (paragraph [0771]). Gibbons teaches a POC device capable of performing multiplexed assays on a small sample would be desirable (paragraph [0004]).
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 performing a particle-based immunoassay of modified Dou to incorporate the teachings of the teachings of image processing algorithms to analyze the morphological operations and shape analysis of objects, and the use of colored compounds for detecting a molecule of Gibbons (paragraphs [0413], [0614],[0681],[0726]) and the teachings of parallel processing and imaging of different assays of Gibbons (paragraphs [0603],[0771]) to provide: wherein the performing of the particle-based immunoassay on a flowing fluid sample that is flowing through a channel comprises performing at least two separate particle based immunoassays, wherein each of the separate particle based immunoassays, provides a separate optical property of morphological and/or color characteristic of aggregates formed by said particles, and wherein the performing image analysis of the obtained images comprises analyses of each separate optical property of morphological and/or color characteristic of aggregates formed by said particles in said separate immunoassays, by said fluid analyzer. Doing so would have a reasonable expectation of successfully allowing for multiplexing of assays on a sample as desired by Gibbons (paragraph [0004]), and improving characterization and analysis of the aggregation and thus allow for determining of a concentration of the aggregation as taught by Gibbons (paragraph [0614]). Additionally, doing so would have a reasonable expectation of successfully improving efficiency, portability, and usability for point-of-care testing of characteristics of multiple antigens or analytes in a sample as desired by Dou (paragraph [0071]).
Regarding claim 30, Dou further teaches wherein the immunoassay is performed to determine the concentration of the at least one target analyte in the flowing fluid sample (paragraph [0073] and Fig. 5 teaches the optical imaging system and software determines the target analyte concentration of the sample via the bead and detection probe, wherein paragraph [0050] teaches the images were captured of the fluid sample flowing through the detection window of the microfluidic chip).
While Dou teaches a sample can include bodily fluid, such as whole blood (paragraph [0029]) and assaying for microparticulates, such as proteins and enzymes (paragraph [0027]), and blood cell analysis that detects diseases and pathogens (paragraph [0028]), modified Dou fails to teach: wherein the at least one target analyte is at least one selected from the group consisting of a C-reactive protein (CRP), HbA1C, procalcitonin (PCT), brain natriuretic peptide (BNP), and a combination thereof.
Gibbons teaches analyte that can be detected can include markers associated with diseases (paragraph [0420]) and biomarkers of interest of the heart (paragraph [0421]), wherein the markers include CRP (paragraphs [0425],[0428]) and BNP (paragraph [0428]). Gibbons teaches inflammation markers include CRP (paragraph [0436]). Gibbons teaches markers can also include procalcitonin (paragraph [0441]).
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 at least one target analyte of modified Dou to incorporate the teachings of various markers and biomarkers of interest for diseases, such as CRP, procalcitonin, and BNP of Gibbons (paragraphs [0420],[0421],[0425],[0428],[0436],[0441]) to provide: wherein the at least one target analyte is at least one selected from the group consisting of a C-reactive protein (CRP), HbA1C, procalcitonin (PCT), brain natriuretic peptide (BNP), and a combination thereof. Doing so would have a reasonable expectation of successfully improving analysis and characterization of a blood sample for known and desired markers of diseases and infections as taught by Gibbons.
Claim 26 is rejected under 35 U.S.C. 103 as being unpatentable over Dou in view of Gibbons, Cheng, Bransky, and Oku as applied to claim 25 above, and further in view Iversen et al. (US 20170356834 A1) and Berliner (US 20020001402 A1).
Regarding claim 26, modified Dou fails to teach wherein the intact cells are excluded by a technique comprising: processing an image of the intact cells to produce a background threshold; processing an image of the flowing fluid sample comprising the intact cells and one or more aggregates; and normalizing the image of the flowing fluid sample against the background threshold, thereby excluding intact cells from the image analysis of the flowing fluid sample.
Iversen teaches a method for quantitative detection of particles in fluid (abstract). Iversen teaches background signal and individual groups of cells are excluded from images to improve evaluation of the sensor signal (paragraph [0045]). Iversen teaches an advantage that individual, greatly contaminated parts of the sample carrier window can be excluded in a targeted manner with the detection, and this also includes larger particles, such as air bubbles or particles in a size magnitude (paragraph [0018]). Iversen teaches fading out the background or background noise to simplify and improve the particle detection (paragraph [0004]).
Berliner teaches a system for generating a profile of particulate components of a body fluid sample, wherein a device causes controlled flow of the body fluid sample and the sample is imaged (abstract). Berliner teaches an image of a control blood sample was taken where most of the red blood cells exist in a non-aggregated state, i.e. intact cells (paragraph [0153]). Berliner teaches the image can be analyzed to exclude presence of proteins (paragraph [0153]). Berliner teaches an image acquired from a blood sample taken during the process of aggregation (paragraph [0154]). Berliner teaches comparing control samples with samples taken from individuals suffering from sepsis (paragraph [0160]). Berliner teaches normalizing using a threshold (paragraph [0168] teaches an image is binarized using a threshold), wherein blobs that are greater than and smaller than thresholds are rejected (paragraphs [0171], [0174],[0175]).
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 modified Dou to incorporate the teachings of comparing a control blood sample image of intact cells with an image of a sample with aggregation of Berliner (paragraphs [0153],[0154],[0160]), the teachings of normalizing an image using a threshold to exclude elements of an image of Berliner (paragraphs [0168], [0171],[0174]-[0175]), and the teachings of background signal and individual groups of cells are excluded from images to improve evaluation of the sensor signal of Iversen (paragraphs [0004],[0018], [0045]) to provide: wherein the intact cells are excluded by a technique comprising: processing an image of the intact cells to produce a background threshold; processing an image of the flowing fluid sample comprising the intact cells and one or more aggregates; and normalizing the image of the flowing fluid sample against the background threshold, thereby excluding intact cells from the image analysis of the flowing fluid sample. Doing so would have a reasonable expectation of successfully improving distinguishing, detecting, and analysis of the aggregates in a sample that comprises additional components, such as cells in a blood sample.
Response to Arguments
Applicant’s arguments, see page 12, filed 12/15/2025, with respect to the claim objections and the rejections under 35 U.S.C. 112(b) have been fully considered and are persuasive. The claim objections and the rejections under 35 U.S.C. 112(b) of 06/13/2025 have been withdrawn.
Applicant's arguments, see pages 13-14, filed 12/15/2025, with respect to the rejections under 35 U.S.C. 101 have been fully considered but they are not persuasive.
In response to applicant’s argument that the claims are not directed to mental steps, are integrated into a practical application, and provides a technical improvement over the art of fluid analysis (Remarks, pages 13-14), the examiner disagrees.
In response to applicant’s argument that the claims cannot be done using a mental process or mathematical calculation (Remarks, page 13), the examiner disagrees. The limitations of “determining a concentration of at least one target analyte in the fluid sample” covers performance of a limitation in the mind, i.e. mental process or mathematical calculation. Regarding the limitations of “determining a concentration”, the instant specification, page 28, lines 21-30 discusses an expression for calculating concentration, which could be performed mentally or by math. Other than “controller circuitry”, if the claim limitations, under its broadest reasonable interpretation, covers performance of the limitations in the mind but for the recitation of generic computer components, then the claim limitations fall within the “Mental Processes” grouping of abstract ideas (MPEP 2106.05(f)). Accordingly, the claims recite abstract ideas (Step 2A: Prong 1: Yes). The claim does recite “performing a particle-based immunoassay on a flowing fluid sample that is flowing through a channel; …providing…imaging…performing image analysis…”, however these steps are data gathering steps, wherein data gathering to be used in the abstract idea is an insignificant extra-solution activity, and not a practical application (see MPEP 2106.05(g)). Thus, the claims are directed to an abstract idea that is not integrated into a practical application (Step 2A, Prong 2: No).
In response to applicant’s argument that the claim as a whole which includes imaging of the flowing fluid sample are not mere abstract data collection (Remarks, page 14), the examiner disagrees. The claim 1 recite “performing a particle-based immunoassay on a flowing fluid sample that is flowing through a channel; providing a fluid analyzer…imaging…performing image analysis…”, however these steps are data gathering steps for the abstract idea within step (d)(ii), wherein data gathering to be used in the abstract idea is an insignificant extra-solution activity, and not a practical application (see MPEP 2106.05(g)). Additionally, the claim does recite a “controller circuitry”, however, the controller circuitry is recited at a high-level of generality (i.e., as generic application operating on a generic computer) such that it amounts no more than mere instructions to apply the exception using a generic computer component; wherein a general purpose computer is not a particular machine (MPEP 2106.05(b)). The steps of imaging analysis of particles are well-understood, routine and conventional activities as evidenced by at least Dou et al. (US 20170343466 A1), Gibbons et al. (US 20120309636 A1; cited in the OA filed 07/26/2024), Putnam et al. (US 20150087559 A1), Cheng et al. (US 20160334396 A1; cited in the IDS filed 06/01/2023), Iversen et al. (US 20170356834 A1), Berliner (US 20020001402 A1), Bransky et al. (US 20160199834 A1) and Oku et al. (US 6106778 A). Additionally, applicant discusses that determining a concentration of an analyte is a well known process (Remarks, page 14). See MPEP 2106.05(d). Therefore, the additional elements of the claims and dependent claims do not comprise an inventive concept when considered individually or as an ordered combination that transforms the claimed judicial exception into a patent-eligible application of the judicial exception. Therefore, the claims do not amount to significantly more than the judicial exception itself (Step 2B: No). The claims are not patent eligible.
Applicant’s arguments, see pages 15-17, filed 12/15/2025, with respect to the rejections under 35 U.S.C. 103, specifically regarding claim 1, have been fully considered and are persuasive. Therefore, the rejection has been withdrawn. However, upon further consideration, a new ground(s) of rejection is made in view of Dou et al. (US 20170343466 A1) in view of Gibbons et al. (US 20120309636 A1; cited in the OA filed 07/26/2024) and Cheng et al. (US 20160334396 A1; cited in the IDS filed 06/01/2023).
Conclusion
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure.
Poher et al. (US 20130323757 A1) teaches a method for characterizing a variation in speed of particles or agglomeration of particles (abstract). Poher teaches one will understand that it is possible to observe, or even quantify, an agglutination state of particles in a biological fluid, using indicators relative to the image obtained by lenseless imaging; and agglutination state depends on the concentration of an analyte in the biological liquid, the quantification of that agglutination state then making it possible to assay that analyte in the liquid (paragraph [0251]).
Chou et al. (US 20210255177 A1; effectively filed 08/16/2018) teaches devices and methods for performing biological and chemical assays (abstract). Chou teaches the sample is imaged, and the particle aggregations in the image are counted, sized and analyzed to measure the concentration of a target analyte (paragraph [0078]). Chou teaches imaging of aggregation particles, where aggregation size clearly correlates with analyte concentration (paragraph [0127]).
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to HENRY H NGUYEN whose telephone number is (571)272-2338. The examiner can normally be reached M-F 7:30A-5:00P.
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.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Maris Kessel can be reached at (571) 270-7698. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/HENRY H NGUYEN/Primary Examiner, Art Unit 1758