DETAILED ACTION
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
Applicant’s Amendments filed on November 07, 2025, has been entered and made of record.
Currently Pending Claim(s) 1-29
Independent Claim(s) 1 and 19
Amended Claim(s) 14
Response to Arguments
This office action is responsive to Applicant’s Arguments/Remarks made in an amendment received on November 07, 2025.
In view of amendments, the Applicant has submitted updated drawing sheets. Figures 3, 6, and 15 have been updated by adding descriptive text labels to the rectangles in each flowchart; thus, the updated drawing sheets overcome the objection to the drawings in the non-final rejection (dated July 14, 2025). Additionally, the Applicant has amended claim 14 to remove the redundancy of listing several thickness ranges which overlap; thus, the objection to the claim 14 has been overcome.
The claims have not been amended regarding the previous rejections under 35 USC 103, but rather, the Applicant argues the rationale of the rejection applied by the Examiner. Upon reviewing the Applicant’s argument, the Examiner finds the argument to be persuasive.
Firstly, the Applicant argued (on Remarks pages 5-6) that the rejection to claim 1, regarding the use of a fixative, is improper. The Applicant showed that the quoted paragraph [0055] from Bonnet (WO 2019/102422 A1) teaches examining and sorting fish eggs based on transparency, and the quoted paragraph [Methods, Par. 1] from Ma (Transcriptome analysis of egg viability in rainbow trout, Oncorhynchus mykiss. BMC Genomics 20, 319.) teaches using a fixative to increase the transparency of fish eggs for analyzing cell cleavage stages after fertilization. The Applicant then argued that because Bonnet teaches sorting based on transparency, combining these references is improper since using a fixative would interfere with the transparency. However, the Examiner points out that the quoted paragraph from Bonnet is one embodiment, and that Bonnet teaches an apparatus for sorting fish eggs based on different optical parameters. In [0023] and [0042], Bonnet teaches sorting based on morphology instead of transparency ([0023] “In one embodiment, the optical parameter of said biological entity comprises transparency, reflectance, contour, morphology, fluorescence or any combination of the foregoing.” [0042] “A simple embodiment of the system of the invention relies in the device as disclosed coupled with a camera as a detection means for e.g. assessing the morphology or the contour of the biological entity 1000: in this alternative, even the sunlight can be a suitable illumination means. Another simple alternative is the coupling of a LED light source with a photodiode sensor for e.g. assessing the transparency/opacity of the biological entity 1000.”). Therefore, as shown by Ma [Methods, Par. 1], it would be obvious to one of ordinary skill in the art to utilize a fixative when examining fish eggs for cell division, which is within the category of morphology.
Next, the Applicant argued (on Remarks pages 6-7) that the 35 USC 103 rejection to claim 1 using Boerjan (US 6,149,956) to teach capturing at least one image depicting the blastodisc of the egg is improper. Bonnet teaches sorting based on an optical parameter, which can be morphology [0042], but Bonnet does not specifically mention imaging the blastodisc. The Examiner had shown that Boerjan teaches NMR imaging the blastodisc of a chicken egg for selecting fertilized eggs. However, the Applicant argued that Bonnet teaches analyzing an optical parameter while Boerjan does not, since Boerjan utilizes NRM imaging. The Examiner finds this argument to be persuasive. The observation of an optical parameter is imperative to the claimed invention, and it provides a reasoning for the use of a fixative, which would be unnecessary in a NMR imaging system. Thus, the Examiner agrees that the combination of Bonnet and Boerjan does not effectively teach the limitation “analysing the at least one image for detecting divided cells in the blastodisc of the egg; and identifying each egg associated with an image showing divided cells in the blastodisc as a fertilized egg,” of claim 1.
However, upon further search and consideration, the prior art of Hisaoka (The normal developmental stages of the zebrafish, brachydanio rerio (hamilton-buchanan). Journal of Morphology. 102:311-327.) was found, and Hisaoka teaches optical imaging of a fish egg for observing cell division in the blastodisc. Additionally, Hisaoka’s observation method utilizes the transparency of zebrafish eggs along with a fixative for observing the blastodisc ([Hisaoka page 312, Materials and Methods] “The following descriptions of the developmental stages were made from studies of the gross specimens together with serial sections of embryos. The latter were previously fixed in Bouin's fluid, embedded in Tissuemat, sectioned at 1011 and stained with haematoxylin and eosin.”), so Hisaoka better applies to the concerns of the Applicant in the arguments above. Therefore, a new rejection was applied to the claims, and this action is non-final. Additionally, the Examiner applies interpretation under 35 USC 112(f) and rejections under 35 USC 101 that should have been included within the first non-final rejection (dated July 14, 2025).
Claim Interpretation
The following is a quotation of 35 U.S.C. 112(f):
(f) Element in Claim for a Combination. – An element in a claim for a combination may be expressed as a means or step for performing a specified function without the recital of structure, material, or acts in support thereof, and such claim shall be construed to cover the corresponding structure, material, or acts described in the specification and equivalents thereof.
The claims in this application are given their broadest reasonable interpretation using the plain meaning of the claim language in light of the specification as it would be understood by one of ordinary skill in the art. The broadest reasonable interpretation of a claim element (also commonly referred to as a claim limitation) is limited by the description in the specification when 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, is invoked.
As explained in MPEP § 2181, subsection I, claim limitations that meet the following three-prong test will be interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph:
(A) the claim limitation uses the term “means” or “step” or a term used as a substitute for “means” that is a generic placeholder (also called a nonce term or a non-structural term having no specific structural meaning) for performing the claimed function;
(B) the term “means” or “step” or the generic placeholder is modified by functional language, typically, but not always linked by the transition word “for” (e.g., “means for”) or another linking word or phrase, such as “configured to” or “so that”; and
(C) the term “means” or “step” or the generic placeholder is not modified by sufficient structure, material, or acts for performing the claimed function.
Use of the word “means” (or “step”) in a claim with functional language creates a rebuttable presumption that the claim limitation is to be treated in accordance with 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph. The presumption that the claim limitation is interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, is rebutted when the claim limitation recites sufficient structure, material, or acts to entirely perform the recited function.
Absence of the word “means” (or “step”) in a claim creates a rebuttable presumption that the claim limitation is not to be treated in accordance with 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph. The presumption that the claim limitation is not interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, is rebutted when the claim limitation recites function without reciting sufficient structure, material or acts to entirely perform the recited function.
Claim limitations in this application that use the word “means” (or “step”) are being interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, except as otherwise indicated in an Office action. Conversely, claim limitations in this application that do not use the word “means” (or “step”) are not being interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, except as otherwise indicated in an Office action.
This application includes one or more claim limitations that do not use the word “means,” but are nonetheless being interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, because the claim limitation(s) uses a generic placeholder that is coupled with functional language without reciting sufficient structure to perform the recited function and the generic placeholder is not preceded by a structural modifier. Such claim limitations are:
Imaging System in claims 10, 19, 21, 24, 26, 27, 28.
Image Analysis System in claim 19 and 29.
Conveyor System in claim 19 and 24 (In claim 25, sufficient structure is provided).
Fixation System in claim 20.
Machine Learning System in claim 29.
Because these claim limitations are being interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, they are being interpreted to cover the corresponding structure described in the specification as performing the claimed function, and equivalents thereof.
An Imaging System is interpreted as comprising a camera mounted for capturing images of an egg as it passes in a conduit (Specification page 7, line 25-28).
An Image Analysis System is interpreted as a generic computer capable of running a machine learning algorithm (Specification page 9, lines 3-10). The computer includes a CPU, memory, and I/O devices such as a computer screen (Specification page 14, lines 14-18).
A Conveyor System is interpreted as a funnel followed by a conduit which is only wide enough to allow eggs to pass through single-file (Specification page 7, lines 8-24).
A Fixation System is interpreted as any container capable of holding eggs and fixative, allowing for the two to mix (Specification page 6, line 32-34 and Fig. 1 label 106).
A Machine Learning System is interpreted as a machine learning algorithm trained to classify the stages of embryonic development from images (Specification page 16, lines 22-25).
If applicant does not intend to have these limitations interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph, applicant may: (1) amend the claim limitation(s) to avoid them being interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph (e.g., by reciting sufficient structure to perform the claimed function); or (2) present a sufficient showing that the claim limitation(s) recite(s) sufficient structure to perform the claimed function so as to avoid it/them being interpreted under 35 U.S.C. 112(f) or pre-AIA 35 U.S.C. 112, sixth paragraph.
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.
Claim Interpretation: Under the broadest reasonable interpretation, the terms of the claim are presumed to have their plain meaning consistent with the specification as it would be interpreted by one of ordinary skill in the art. See MPEP 2111.
Claims 1 and 19 are directed to non-statutory subject matter.
Claims 1 and 19 do not fall within at least one of the four categories of patent eligible subject matter. In these claims, several steps are recited:
analyzing the at least one image for detecting divided cells in the blastodisc of the egg
identifying each egg associated with an image showing divided cells in the blastodisc as a fertilized egg
Step 1: This part of the eligibility analysis evaluates whether the claim falls within any statutory category. See MPEP 2106.03. The claims recite a system and a method that implements an AI module, and falls within one of the statutory categories of invention.
The broadest reasonable interpretation of steps (a), (b), and (c) is that those steps fall within the mental process groupings of abstract ideas because they cover concepts performed in the human mind, including observation, evaluation, judgment, and opinion. See MPEP 2106.04(a)(2), subsection III.
Specifically, Step (a) recites analyzing an image, which may be practically performed in the human mind by using observation, evaluation, and judgement. Step (b) recites identifying objects in an image based on an observable optical parameter, which may also be practically performed in the human mind. As such, a person could mentally observe captured images and select eggs which exhibit cell division in the images. The mere nominal recitation that the various steps are being executed using a camera does not take the limitations out of the mental process grouping. Thus, the claims recite a mental process.
Step 2A, Prong Two: This part of the eligibility analysis evaluates whether the claim as a whole integrates the recited judicial exception into a practical application of the exception or whether the claim is “directed to” the judicial exception. This evaluation is performed by (1) identifying whether there are any additional elements recited in the claim beyond the judicial exception, and (2) evaluating those additional elements individually and in combination to determine whether the claim as a whole integrates the exception into a practical application. See MPEP 2106.04(d). Claim 1 recites the additional elements of:
within a period of time from initial fertilization of the batch, treating the sample with a fixative bringing an outer membrane of the eggs at least partly transparent and/or a blastodisc of the eggs at least partly opaque
capturing at least one image of individual eggs of the sample depicting the blastodisc of the egg
Additional element (a) This limitation refers to an insignificant pre/post-solution extra activity that does not add a meaningful limitation to the abstract idea because it amounts to simply implementing the abstract idea in a method. Additional element (b) involves capturing an image using a camera, which is generic computer components and/or insignificant pre/post-solution extra activity that does not add a meaningful limitation to the abstract idea because it amounts to simply implementing the abstract idea in a method.
Step 2B: This part of the eligibility analysis evaluates whether the claim as a whole amount to significantly more than the recited exception i.e., whether any additional element, or combination of additional elements, adds an inventive concept to the claim. See MPEP 2106.05.
Even when considered in combination, the additional element represents mere instructions to implement an abstract idea or other exception using a camera and insignificant extra-solution activity, which do not provide an inventive concept.
The above reasoning is applied to both claims 1 and 19. As shown in the 35 USC 112(f) discussion above, claim 19 lacks the sufficient structure necessary for the method to be integrated into a practical application.
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.
Claims 1, 10-12, 19-20, and 25 are rejected under 35 U.S.C. 103 as being unpatentable over Bonnet et al. (WO 2019/102422 A1), hereafter Bonnet, further in view of Ma et al. (Transcriptome analysis of egg viability in rainbow trout, Oncorhynchus mykiss. BMC Genomics 20, 319), hereafter Ma, and Hisaoka et al. (The normal developmental stages of the zebrafish, brachydanio rerio (hamilton-buchanan). Journal of Morphology. 102:311-327.), hereafter Hisaoka.
Regarding claim 1, Bonnet teaches a method of identifying fertilized fish eggs in a sample of eggs taken from a batch of incubating fish eggs ([0053] “In a particular embodiment, the detecting and analysing system 300 comprising a camera 301 is adapted to operate the selector 200 on the basis of transparency of zebrafish larvae, so to sort fertilized from non-fertilized egg cells. In this way, fertilized egg cells can be dispensed into a dedicated reservoir 500 whereas non-fertilized egg cells could be eliminated from further analysis upon disposal into a waste reservoir 501.”),
Bonnet fails to teach treating the sample with a fixative. However, Ma teaches within a period of time from initial fertilization of the batch, treating the sample with a fixative bringing an outer membrane of the eggs at least partly transparent and/or a blastodisc of the eggs at least partly opaque ([Methods Par. 1] “The eggs were incubated at 10 °C for about the first 24 h post fertilization, after which time a sample of about 25–60 embryos were collected from each family and fixed in Stockard’s solution to evaluate early embryonic survival and viability at about the 32-cell stage by enumerating the embryos reaching each stage of cell cleavage (Table 1).”);
Bonnet and Ma are analogous in the art because both teach methods of examining fish eggs through image capturing methods. Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by utilizing a fixative to make the eggs transparent. This modification allows for the eggs to be transparent when imaging the cell cleavage of the embryo ([Methods Par. 1] “…to evaluate early embryonic survival and viability at about the 32-cell stage by enumerating the embryos reaching each stage of cell cleavage.”).
Additionally, Bonnet fails to teach capturing at least one image of the individual eggs of the sample depicting the blastodisc of the egg, analysing the at least one image for detecting divided cells in the blastodisc of the egg, and identifying each egg associated with an image showing divided cells in the blastodisc as a fertilized egg.
However, Hisaoka teaches capturing at least one image of individual eggs of the sample depicting the blastodisc of the egg ([Page 319, Summary] “1. Twenty-five selected developmental stages of the zebra-fish, Brachydmio rerio are described. Each stage is illustrated by one or more photomicrographs taken with the phase contrast microscope.” See the images on page 323.);
analysing the at least one image for detecting divided cells in the blastodisc of the egg; and identifying each egg associated with an image showing divided cells in the blastodisc as a fertilized egg ([Page 319, Summary] “2. Stage 1 represents the recently fertilized ovum, and stages 2 to 7 the formation of the blastodisc and the initial cleavage divisions. Ova composed of 64 to 256 blastomeres are designated as late cleavage stages (8).” On page 313, Hisaoka shows the stages of development after fertilization, and the stages are indicated by cell division and the amount of time passed since fertilization. Thus, one of ordinary skill in the art would be expected to conclude that cell division over time indicates that an egg is fertilized.).
Bonnet and Hisaoka are analogous in the art to the claimed invention, because both teach methods of observing an optical parameter of fish eggs. Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to identify fertilized eggs based on observed cell division in optical images. This modification would apply well-known methods of observing cell-division as an optical parameter for determining fertility and growth stages. Bonnet specifically motivates doing so by teaching that eggs can be sorted based on an observed optical parameter of morphology of an embryo, egg cell, etc. (Bonnet [0018] “In one embodiment, the egg cell, the zygote, the embryo or the larva is derived from an insect, an amphibian or a fish. In one embodiment, the means for detecting and analysing an optical parameter of said biological entity comprises an optical sensor and a light source. In one embodiment, the optical parameter of said biological entity comprises transparency, reflectance, contour, morphology, fluorescence or any combination of the foregoing.”), and Bonnet teaches that a solution for high-throughput sorting based on fertility is needed for assuring that non-fertilized eggs do not contaminate fertilized eggs ([0006] “the current bottleneck for high-throughput screening is an efficient sorting and dispensing process of individual organisms into single reservoirs such as wells of a multiwell plate. One of the main tasks for the people working with zebrafish in laboratories is to take care of their reproduction and egg sorting, because the eggs that will not produce larvae might contaminate the fertilized ones. This monotonous task as of today is performed manually and is very time-consuming, prone to errors and complicated due to the small dimensions of the eggs (~1 mm diameter).”).
Regarding claim 10, Bonnet teaches wherein said step of capturing at least one image of individual eggs comprises bringing the eggs, one-by-one, passed an imaging system (Fig. 1 shows the biological entities passing the optical sensor 300 one-by-one. [0042] “…means 300 for detecting and analysing an optical parameter of said biological entity 1000 is located. In particular, said means 300 comprise an optical sensor and a light source adapted to reveal an optical parameter of the biological entity 1000 such as transparency or opacity, reflectance, contour, morphology, fluorescence or any combination of the foregoing.”).
Regarding claim 11, Bonnet fails to teach wherein said period of time from initial fertilization of the batch to treating the sample with the fixative is within the range of 30 to 220 degree-hours. However, Ma teaches treating the sample with a fixative at about 240 degree-hours post fertilization ([Methods Par. 1] “The eggs were incubated at 10 °C for about the first 24 h post fertilization, after which time a sample of about 25–60 embryos were collected from each family and fixed in Stockard’s solution to evaluate early embryonic survival and viability at about the 32-cell stage by enumerating the embryos reaching each stage of cell cleavage (Table 1).”).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by treating the eggs with a fixative in the range of 30-220 degree-hours, because such a modification would have been obvious to try. More specifically, Ma teaches that about 240 degree hours is the time to reach the 32-cell stage of embryonic development for rainbow trout eggs. Therefore, it would have obvious to try to modify Bonnet’s invention to include a range of degree-hours lower than 240 since there are a finite number of identified, predictable potential solutions to the recognized need (as discussed above) and one of ordinary skill in the art could have pursued the known potential solutions with a reasonable expectation of success.
Regarding claim 12, Bonnet fails to teach a fixative comrpising any one of: an acetic acid-alcohol-formaldehyde solution (Serra's fixative) Stockards solution, or AquaBoost® Quattro. However, Ma teaches wherein said fixative comprises any one of: an acetic acid-alcohol-formaldehyde solution (Serra's fixative) Stockards solution, or AquaBoost® Quattro ([Methods Par. 1] “The eggs were incubated at 10 °C for about the first 24 h post fertilization, after which time a sample of about 25–60 embryos were collected from each family and fixed in Stockard’s solution to evaluate early embryonic survival and viability at about the 32-cell stage by enumerating the embryos reaching each stage of cell cleavage (Table 1).”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by utilizing Stockard’s solution to make the eggs transparent. This modification allows for the eggs to be transparent when imaging the cell cleavage of the embryo ([Methods Par. 1] “…to evaluate early embryonic survival and viability at about the 32-cell stage by enumerating the embryos reaching each stage of cell cleavage.”).
Regarding claim 19, Bonnet teaches a fish egg analysis system ([0001] “…the present invention relates to an apparatus for use as a sorter for biological entities such as embryos or egg cells.”) wherein:
at least one imaging system being configured for capturing at least one image of individual fish eggs of a sample of eggs comprising fertilized and unfertilized fish eggs ([0053] “In a particular embodiment, the detecting and analysing system 300 comprising a camera 301 is adapted to operate the selector 200 on the basis of transparency of zebrafish larvae, so to sort fertilized from non-fertilized egg cells.”) and a
a conveyor system arranged for conveying the eggs of the sample past the at least one imaging system in single file (See Fig. 1 showing a channel which causes the eggs to pass the camera in single file.).
Although Bonnet teaches at least one imaging system for capturing at least one image of individual fish eggs, Bonnet fails to teach the at least one image depicting the blastodisc of the eggs. However, Hisaoka teaches depicting the blastodisc of the eggs ([Page 319, Summary] “1. Twenty-five selected developmental stages of the zebra-fish, Brachydmio rerio are described. Each stage is illustrated by one or more photomicrographs taken with the phase contrast microscope.” See the images on page 323.”);
Additionally, although Bonnet teaches an image analysis means ([Abstract] “Means for detecting and analysing an optical parameter of the biological entities is coupled with the selector.”), Bonnet fails to teach the image analysis system being configured for analyzing the captured images for detecting divided calls in the blastodisc and identifying each egg associated with an image showing divided cells in the blastodisc as a fertilized egg. However, Hisaoka teaches an image analysis system being configured for analysing the captured images for detecting divided cells in the blastodisc and identifying each egg associated with an image showing divided cells in the blastodisc as a fertilized egg ([Page 319, Summary] “1. Twenty-five selected developmental stages of the zebra-fish, Brachydmio rerio are described. Each stage is illustrated by one or more photomicrographs taken with the phase contrast microscope.” See the images on page 323.).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to identify fertilized eggs based on observed cell division in optical images. This modification would apply well-known methods of observing cell-division as an optical parameter for determining fertility and growth stages. Bonnet specifically motivates doing so by teaching that eggs can be sorted based on an observed optical parameter of morphology of an embryo, egg cell, etc. (Bonnet [0018] “In one embodiment, the egg cell, the zygote, the embryo or the larva is derived from an insect, an amphibian or a fish. In one embodiment, the means for detecting and analysing an optical parameter of said biological entity comprises an optical sensor and a light source. In one embodiment, the optical parameter of said biological entity comprises transparency, reflectance, contour, morphology, fluorescence or any combination of the foregoing.”), and Bonnet teaches that a solution for high-throughput sorting based on fertility is needed for assuring that non-fertilized eggs do not contaminate fertilized eggs ([0006] “the current bottleneck for high-throughput screening is an efficient sorting and dispensing process of individual organisms into single reservoirs such as wells of a multiwell plate. One of the main tasks for the people working with zebrafish in laboratories is to take care of their reproduction and egg sorting, because the eggs that will not produce larvae might contaminate the fertilized ones. This monotonous task as of today is performed manually and is very time-consuming, prone to errors and complicated due to the small dimensions of the eggs (~1 mm diameter).”).
Regarding claim 20, Bonnet fails to teach a fixation system. However, Ma teaches wherein a fixation system configured for mixing a sample of eggs comprising fertilized and unfertilized fish eggs with a fixative bringing an outer membrane of the eggs at least partly transparent and/or a blastodisc of the eggs at least partly opaque ([Methods Par. 1] “The eggs were incubated at 10 °C for about the first 24 h post fertilization, after which time a sample of about 25–60 embryos were collected from each family and fixed in Stockard’s solution to evaluate early embryonic survival and viability at about the 32-cell stage by enumerating the embryos reaching each stage of cell cleavage (Table 1).”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by utilizing a fixative to make the eggs transparent. This modification allows for the eggs to be transparent when imaging the cell cleavage of the embryo ([Methods Par. 1] “…to evaluate early embryonic survival and viability at about the 32-cell stage by enumerating the embryos reaching each stage of cell cleavage.”).
Regarding claim 25, Bonnet teaches wherein the conveyor system comprises a tubular conduit for conveying the eggs, the conduit having an inner diameter D within the range of Degg< D < 2*Degg, where Degg is the diameter of the eggs of the sample (Fig. 1 shows the biological entities passing the optical sensor 300 one-by-one in a channel.).
Claims 2-9, 21-23, and 26-27 are rejected under 35 U.S.C. 103 as being unpatentable over Bonnet, Ma, and Hisaoka, further in view of Sebesta (CN 102460124 A).
Regarding claim 2, Bonnet fails to teach capturing a plurality of images of each egg from a plurality of directions. However, Sebesta teaches wherein said step of capturing at least one image of individual eggs comprises capturing a plurality of images of each egg from a plurality of directions ([0094] “According to one embodiment of the method of the invention, analyzed images of the egg or embryo taken from different angles are combined by a tomographic algorithm for generating a true three-dimensional reconstruction.”).
Bonnet and Sebesta are analogous in the art because both teach methods for analyzing an egg for fertility using a camera and image analysis. Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by capturing a plurality of images from different angles when observing and analyzing the egg’s fertility for sorting. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 3, Bonnet fails to teach wherein capturing at least one image of individual eggs comprises capturing a first set of images of each individual egg when rotated in a first plane and capturing a second set of images of the individual egg when rotated in a second plane. However, Sebesta teaches wherein said step of capturing at least one image of individual eggs comprises capturing a first set of images of each individual egg when rotated in a first plane and capturing a second set of images of the individual egg when rotated in a second plane ([0092] “Another way to increase the information collected is to take several images from different angles. If the egg or embryo is rotated at a constant rotational speed and images are taken at specific time intervals, the images can be used for tomographic reconstruction of the embryo. Alternatively, the egg or embryo is rotated a specific angle at a time and an image is taken after each rotation. The target is rotated about an axis that is not parallel to the target beam. Preferably, the target rotates about an axis perpendicular to the target beam.”).
Therefore, it would have been obvious to one of ordinary skill in the art to modify Bonnet’s invention by rotating the egg during the imaging process. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 4, Bonnet fails to teach a second plane orthogonal to the first plane. However, Sebesta teaches wherein the second plane is orthogonal to the first plane ([0092] “Another way to increase the information collected is to take several images from different angles. If the egg or embryo is rotated at a constant rotational speed and images are taken at specific time intervals, the images can be used for tomographic reconstruction of the embryo. Alternatively, the egg or embryo is rotated a specific angle at a time and an image is taken after each rotation. The target is rotated about an axis that is not parallel to the target beam. Preferably, the target rotates about an axis perpendicular to the target beam. Rotation of the egg or embryo enables a true three-dimensional reconstruction of the egg or embryo.” Additionally, Sebesta discloses optical orthogonal microscopy in the prior art, which includes imaging an object from orthogonal planes. [0007] “The phase image is generated by interpreting the interferograms from four different cameras, where the reference light passes through a 1/4 wave plate to produce a 90-degree phase shift of the orthogonal P-plane and S-plane polarizations. A polarization beam splitter is then used to separate the signals of the two polarization directions, which are then detected on two separate cameras.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by capturing images from two orthogonal planes for imaging and analysis of each individual egg. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 5, Bonnet teaches wherein capturing the first set of images comprises capturing the first set of images with a first camera having an optical axis located in said first plane (See the camera labeled 300 in Fig. 1. [0016] “…means for detecting and analysing an optical parameter of said biological entity;).
Bonnet fails to teach wherein capturing the second set of images comprises capturing the second set of images with a second camera having an optical axis located in said second plane. However, Sebesta teaches wherein capturing the second set of images comprises capturing the second set of images with a second camera having an optical axis located in said second plane (Sebesta discloses optical orthogonal microscopy in the prior art, which includes imaging an object from orthogonal planes with multiple cameras simultaneously. [0007] “The phase image is generated by interpreting the interferograms from four different cameras, where the reference light passes through a 1/4 wave plate to produce a 90-degree phase shift of the orthogonal P-plane and S-plane polarizations. A polarization beam splitter is then used to separate the signals of the two polarization directions, which are then detected on two separate cameras.”).
Therefore, it would have been obvious for one of ordinary skill in the art to modify Bonnet’s invention by capturing images from a plurality of directions when imaging and analyzing each egg. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 6, Bonnet fails to teach where capturing any one of the first set of images and the second set of images comprises capturing images of said individual egg when it is rotated an entire revolution. However, Sebesta teaches wherein capturing any one of the first set of images and the second set of images comprises capturing images of said individual egg when it is rotated an entire revolution ([0092] “Alternatively, the egg or embryo rotates by a specific angle each time, and the image is taken after each rotation. The target rotates around an axis that is not parallel to the target beam. Preferably, the target rotates about an axis perpendicular to the target beam. The rotation of an egg or embryo makes it possible to reconstruct the egg or embryo in real three dimensions.” The egg is continuously rotated, and this process can continue as images are captured at multiple time points. [0098] “Under the correct culture conditions, entire developmental processes can be studied by noninvasive time-lapse imaging.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by rotating the egg an entire revolution when imaging from multiple directions. This modification would allow for the maximum amount of angles and information to be collected when observing the egg; additionally, observing the egg over time allows for meaningful developmental changes to be analyzed (Sebesta [0097] The two methods, that is, using at least two wavelengths or taking images from different angles, can also be used in combination to maximize the information collected.” [0099] “By acquiring images at different time points or continuously, the development of eggs or embryos can be studied. For example, it would be meaningful to study possible changes in eggs, such as changes in zona pellucida when fertilization occurs.”).
Regarding claim 7, Bonnet fails to teach where capturing any one of the first set of images and the second set of images comprises rotating said individual egg between image captures within the range of 45° to 135°. However, Sebesta teaches wherein capturing any one of the first set of images and the second set of images comprises rotating said individual egg between image captures within the range of 45° to 135° ([0092] “Another way to increase the information collected is to take several images from different angles. If the egg or embryo is rotated at a constant rotational speed and images are taken at specific time intervals, the images can be used for tomographic reconstruction of the embryo. Alternatively, the egg or embryo is rotated a specific angle at a time and an image is taken after each rotation. The target is rotated about an axis that is not parallel to the target beam. Preferably, the target rotates about an axis perpendicular to the target beam. Rotation of the egg or embryo enables a true three-dimensional reconstruction of the egg or embryo.” Additionally, Sebesta discloses optical orthogonal microscopy in the prior art, which includes imaging an object from orthogonal planes. [0007] “The phase image is generated by interpreting the interferograms from four different cameras, where the reference light passes through a 1/4 wave plate to produce a 90-degree phase shift of the orthogonal P-plane and S-plane polarizations. A polarization beam splitter is then used to separate the signals of the two polarization directions, which are then detected on two separate cameras.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by capturing images from two orthogonal planes for imaging and analysis of each individual egg. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 8, Bonnet fails to teach where capturing at least one image of individual eggs comprises capturing, in a single exposure, a plurality of images of each egg from a plurality of directions. However, Sebesta teaches wherein said step of capturing at least one image of individual eggs comprises capturing, in a single exposure, a plurality of images of each egg from a plurality of directions (Sebesta discloses optical orthogonal microscopy in the prior art, which includes imaging an object from orthogonal planes with multiple cameras simultaneously. [0007] “The phase image is generated by interpreting the interferograms from four different cameras, where the reference light passes through a 1/4 wave plate to produce a 90-degree phase shift of the orthogonal P-plane and S-plane polarizations. A polarization beam splitter is then used to separate the signals of the two polarization directions, which are then detected on two separate cameras.”).
Therefore, it would have been obvious for one of ordinary skill in the art to modify Bonnet’s invention by capturing images from a plurality of directions when imaging and analyzing each egg. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 9, Bonnet fails to teach using one or a plurality of prisms and/or mirrors to capture, in said single exposure, said images of the egg from said plurality of directions. However, Sebesta teaches a method characterised by using one or a plurality of prisms and/or mirrors to capture, in said single exposure, said images of the egg from said plurality of directions (Fig. 4 shows the light beams, a prism, mirrors, and an image sensor; [0109] “The sensor 500 is arranged to detect the interference pattern caused by the target beam 21 and the reference beam 23. Since the target beam 21 and the reference beam 23 are coherent with each other, they will produce an interference pattern at the sensor 500. In particular, since the at least one target beam 21 passes through the sample 10, the target beam 21 and the reference beam 23 propagate different optical path lengths and thus the interference pattern indicates a phase shift between the target beam 21 and the reference beam 23. The sensor 500 may be, for example, a digital sensor such as a CCD (Charge Coupled Device) or a CMOS (Complementary Metal Oxide Semiconductor) image sensor.” Also see paragraph 0110 discussing reconstructing the image.).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by applying Sebesta’s method of imaging and reconstruction when analyzing an egg for cell division in the blastodisc. This modification allows for a reconstruction image of the egg to be used for evaluation of the egg quality or development stage ([0110] “This information can also be used, for example, to determine the shape and optical density of the at least one egg or embryo 11. In particular, the processing unit 13 is arranged to determine at least one egg or embryo quality representative parameter from said phase and/or amplitude information.”).
Regarding claim 21, Bonnet fails to teach a first camera being configured for capturing a first set of images of each of said individual fish eggs when rotated in a first plane; and a second camera being configured for capturing a second set of images of the individual fish egg when rotated in a second plane. However, Sebesta teaches wherein said at least one imaging system comprises; a first camera being configured for capturing a first set of images of each of said individual fish eggs when rotated in a first plane; and a second camera being configured for capturing a second set of images of the individual fish egg when rotated in a second plane ([0092] “Another way to increase the information collected is to take several images from different angles. If the egg or embryo is rotated at a constant rotational speed and images are taken at specific time intervals, the images can be used for tomographic reconstruction of the embryo. Alternatively, the egg or embryo is rotated a specific angle at a time and an image is taken after each rotation. The target is rotated about an axis that is not parallel to the target beam. Preferably, the target rotates about an axis perpendicular to the target beam. Rotation of the egg or embryo enables a true three-dimensional reconstruction of the egg or embryo.” Additionally, Sebesta discloses optical orthogonal microscopy in the prior art, which includes imaging an object from orthogonal planes. [0007] “The phase image is generated by interpreting the interferograms from four different cameras, where the reference light passes through a 1/4 wave plate to produce a 90-degree phase shift of the orthogonal P-plane and S-plane polarizations. A polarization beam splitter is then used to separate the signals of the two polarization directions, which are then detected on two separate cameras.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by capturing images from two orthogonal planes for imaging and analysis of each individual egg. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 22, Bonnet fails to teach the second plane orthogonal to the first plane. However, Sebesta teaches wherein the second plane is orthogonal to the first plane (Sebesta discloses optical orthogonal microscopy in the prior art, which includes imaging an object from orthogonal planes. [0007] “The phase image is generated by interpreting the interferograms from four different cameras, where the reference light passes through a 1/4 wave plate to produce a 90-degree phase shift of the orthogonal P-plane and S-plane polarizations. A polarization beam splitter is then used to separate the signals of the two polarization directions, which are then detected on two separate cameras.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by capturing images from two orthogonal planes for imaging and analysis of each individual egg. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 23, Bonnet fails to teach the first camera has an optical axis located in said first plane and the second camera has an optical axis located in said second plane. However, Sebesta teaches wherein the first camera has an optical axis located in said first plane and the second camera has an optical axis located in said second plane (Sebesta discloses optical orthogonal microscopy in the prior art, which includes imaging an object from orthogonal planes. [0007] “The phase image is generated by interpreting the interferograms from four different cameras, where the reference light passes through a 1/4 wave plate to produce a 90-degree phase shift of the orthogonal P-plane and S-plane polarizations. A polarization beam splitter is then used to separate the signals of the two polarization directions, which are then detected on two separate cameras.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by capturing images from multiple planes for imaging and analysis of each individual egg. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 26, Bonnet fails to teach capturing images of individual eggs from a plurality of directions. However, Sebesta teaches wherein said at least one imaging system is configured for capturing images of individual eggs from a plurality of directions ([0094] “According to one embodiment of the method of the invention, analyzed images of the egg or embryo taken from different angles are combined by a tomographic algorithm for generating a true three-dimensional reconstruction.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by capturing a plurality of images from different angles when observing and analyzing the egg’s fertility for sorting. This modification allows for the egg to be analyzed from more angles, which increases the information collected and allows for the full three-dimensional shape of the egg to be observed (Sebesta [0092] “Another way to increase the information collected is to take several images from different angles.”).
Regarding claim 27, Bonnet fails to teach teaches one or a plurality of mirrors or prism allowing a camera of the imaging system to capture, in a single exposure, a plurality of images of each egg from different directions. However, Sebesta teaches wherein each of said at least one imaging system comprises one or a plurality of mirrors or prism allowing a camera of the imaging system to capture, in a single exposure, a plurality of images of each egg from different directions (Fig. 4 shows the light beams, a prism, mirrors, and an image sensor; [0109] “The sensor 500 is arranged to detect the interference pattern caused by the target beam 21 and the reference beam 23. Since the target beam 21 and the reference beam 23 are coherent with each other, they will produce an interference pattern at the sensor 500. In particular, since the at least one target beam 21 passes through the sample 10, the target beam 21 and the reference beam 23 propagate different optical path lengths and thus the interference pattern indicates a phase shift between the target beam 21 and the reference beam 23. The sensor 500 may be, for example, a digital sensor such as a CCD (Charge Coupled Device) or a CMOS (Complementary Metal Oxide Semiconductor) image sensor.” Also see paragraph 0110 discussing reconstructing the image.).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by applying Sebesta’s method of imaging and reconstruction when analyzing an egg for cell division in the blastodisc. This modification allows for a reconstruction image of the egg to be used for evaluation of the egg quality or development stage ([0110] “This information can also be used, for example, to determine the shape and optical density of the at least one egg or embryo 11. In particular, the processing unit 13 is arranged to determine at least one egg or embryo quality representative parameter from said phase and/or amplitude information.”).
Claims 13, 15, 18, and 28 are rejected under 35 U.S.C. 103 as being unpatentable over Bonnet, Ma, and Hisaoka, further in view of Zinskie et al. (A mechanical microcompressor for high resolution imaging of motile specimens. Exp Cell Res. 2015), hereafter Zinskie.
Regarding claim 13, Bonnet fails to teach a step of compressing the egg to a predetermined thickness prior to said step of capturing said at least one image of the egg. However, Zinskie teaches wherein by the step of compressing the egg to a predetermined thickness prior to said step of capturing said at least one image of the egg ([Section 1, Par. 2] “Controlled compression of the sample can be obtained with the use of a rotocompressor or a mechanical microcompressor (MMC), a device that allows for manual compression of a specimen between two glass planes.”).
Bonnet and Zinskie are analogous in the art because both teach methods of imaging small biological entities. Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by compressing the egg in a controlled manner before imaging. This modification allows for the egg to be restrained without damaging or distorting the egg beyond recovery (Zinskie [Section 1, Par. 2] “Physical inhibition of a specimen includes its placement in a highly viscous fluid, adherence to, or imbedding in, a surface such as agarose, or uncontrolled squeezing of the sample between a coverslip and slide, often without regard for the amount of distortion. Many times these practices do not affect biochemical processes, however recovery of the specimen can be difficult or impossible.”).
Regarding claim 15, Bonnet fails to teach trapping the egg between two holding elements and bringing the holding elements together compressing the trapped egg. However, Zinskie teaches wherein said step of compressing the egg comprises the sub-steps of: trapping the egg between two holding elements; and bringing the holding elements together compressing the trapped egg ([Section 1, Par. 2] “Controlled compression of the sample can be obtained with the use of a rotocompressor or a mechanical microcompressor (MMC), a device that allows for manual compression of a specimen between two glass planes.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by compressing the egg in a controlled manner before imaging. This modification allows for the egg to be restrained. (Zinskie [Section 1, Par. 2] “Physical inhibition of a specimen includes its placement in a highly viscous fluid, adherence to, or imbedding in, a surface such as agarose, or uncontrolled squeezing of the sample between a coverslip and slide, often without regard for the amount of distortion. Many times these practices do not affect biochemical processes, however recovery of the specimen can be difficult or impossible.”).
Regarding claim 18, Bonnet fails to teach arranging an image-capturing device on a first side of the compressed egg, arranging a light source on a second side of the compressed egg, said second side being opposite the first side, and transmitting light from the light source through the compressed egg and into the image-capturing device.
However, Zinskie teaches wherein said step of capturing an image of the compressed egg comprises the sub-steps off: arranging an image-capturing device on a first side of the compressed egg; arranging a light source on a second side of the compressed egg, said second side being opposite the first side; and transmitting light from the light source through the compressed egg and into the image-capturing device ([Section 3.1, Par. 2] “Again, we were able to gently compress them and imaged them on an OI polarized light microscope that allows real-time birefringence imaging using a video-enhanced polychromatic polscope. We obtained remarkable images (Fig. 1d and e) of trapped cells and also obtained cells contracting their myonemes (see Videos 4 and 5, respectively).” Compressing an object, shining light through the object towards a lens and/or camera, and imaging the object is a well-known use case of utilizing microscopes for microscopic imaging.).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to image the egg using this technique of shining a light through an organism and imaging from the opposite side, allowing light to pass into the camera. This modification uses known techniques in the art, and the compression of the organism allows for the organism to be restrained during observation and imaging (Zinskie [Section 1, Par. 2] “Physical inhibition of a specimen includes its placement in a highly viscous fluid, adherence to, or imbedding in, a surface such as agarose, or uncontrolled squeezing of the sample between a coverslip and slide, often without regard for the amount of distortion. Many times these practices do not affect biochemical processes, however recovery of the specimen can be difficult or impossible.”).
Regarding claim 28, Bonnet fails to teach a pressing device arranged upstream of said at least one imaging system, said pressing device being configured to compress each egg to a predetermined thickness. However, Zinskie teaches wherein a pressing device arranged upstream of said at least one imaging system, said pressing device being configured to compress each egg to a predetermined thickness ([Section 1, Par. 2] “Controlled compression of the sample can be obtained with the use of a rotocompressor or a mechanical microcompressor (MMC), a device that allows for manual compression of a specimen between two glass planes.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by compressing the egg in a controlled manner before imaging. This modification allows for the egg to be restrained without damaging or distorting the egg beyond recovery (Zinskie [Section 1, Par. 2] “Physical inhibition of a specimen includes its placement in a highly viscous fluid, adherence to, or imbedding in, a surface such as agarose, or uncontrolled squeezing of the sample between a coverslip and slide, often without regard for the amount of distortion. Many times these practices do not affect biochemical processes, however recovery of the specimen can be difficult or impossible.”).
Claims 14 is rejected under 35 U.S.C. 103 as being unpatentable over Bonnet, Ma, Hisaoka, and Zinskie, further in view of Weber et al. (Multilayer mounting for long-term light sheet microscopy of zebrafish. Journal of Visualized Experiments. 2014.), hereafter Weber.
Regarding claim 14, Bonnet fails to teach the predefined thickness within the range of 0.1 to 1.5mm. However, Weber teaches, wherein the predefined thickness is any one of: within the range of 0.1 to 1.5 mm; within the range of 0.3 to 1.0 mm; within the range of 0.4 to 0.6 mm; and 0.5 mm ([Imaging of Early Zebrafish Embryogenesis, Par. 1] “The 8hpf zebrafish embryo was mounted with its intact chorion inside an E3-filled tube with an inner diameter of 1mm. That results in a slight clamping of the chorion).
Bonnet and Weber are analogous in the art because both teach methods of imaging fish. Therefore, it would have been obvious to one of ordinary skill before the effective filing date of the claimed invention to mount or clamp the fish eggs within a thickness of 1mm. This modification allows for zebrafish embryos to be mounted and slightly clamped, but the developmental processes are unaffected ([Imaging of Early Zebrafish Embryogenesis, Par. 1] “All developmental processes during embryogenesis are unaffected by the mounting and can be optimally visualized using light sheet microscopy.”).
Claims 16-17 are rejected under 35 U.S.C. 103 as being unpatentable over Bonnet, Ma, Hisaoka, and Zinskie, further in view of Parizat et al. (US 2018/0194913 A1), hereafter Parizat.
Regarding claim 16, Bonnet fails to teach holding elements comprosing of ductile strips of transparent sheet metal. However, Parizat teaches wherein said holding elements comprises ductile strips of transparent sheet material ([Claim 17] “A highly transparent film based on cellulose triacetate, comprising, a support film acting as a support layer comprising cellulose triacetate or a mixture of cellulose esters and cellulose triacetate as a primary component.” [Abstract] “The highly transparent films of the invention can be used as… enclosing film, slide film for microscopy, cover slip replacement film and/or protective film.”).
Zinskie and Parizat are analogous in the art because both teach methods of utilizing slide film in microscopy. Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Zinskie’s invention by using cellulose triacetate (CTA), which is both ductile and clear, as the material for holding the organisms being observed by a microscope and camera. This modification simply applies a method that is well known in the art, which is utilizing CTA as a clear sheet in imaging applications ([0002] “Films based on cellulose derivatives have been produced for more than 100 years. They are also used as substrates for photographic active layers, as support material for membranes or—especially in the last 10 years—as optically inert protective material for (LCD) crystal layers of liquid crystal displays and polarizing layers in LCD screens. Cellulose esters such as cellulose triacetate (CTA), diacetate cellulose (DAC), cellulose acetate butyrate (CAB) or cellulose acetate propionate (CAP) or combinations of these cellulose derivatives are used here as preferred materials.”).
Regarding claim 17, Bonnet fails to teach a transparent sheet material comprising regenerated cellulose. However, Parizat teaches wherein said transparent sheet material comprises regenerated cellulose ([Claim 17] “A highly transparent film based on cellulose triacetate, comprising, a support film acting as a support layer comprising cellulose triacetate or a mixture of cellulose esters and cellulose triacetate as a primary component.” [Abstract] “The highly transparent films of the invention can be used as… enclosing film, slide film for microscopy, cover slip replacement film and/or protective film.”).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Zinskie’s invention by using cellulose triacetate (CTA), which is regenerated cellulose, as the material for holding the organisms being observed by a microscope and camera. This modification simply applies a method that is well known in the art, which is utilizing CTA as a clear sheet in imaging applications ([0002] “Films based on cellulose derivatives have been produced for more than 100 years. They are also used as substrates for photographic active layers, as support material for membranes or—especially in the last 10 years—as optically inert protective material for (LCD) crystal layers of liquid crystal displays and polarizing layers in LCD screens. Cellulose esters such as cellulose triacetate (CTA), diacetate cellulose (DAC), cellulose acetate butyrate (CAB) or cellulose acetate propionate (CAP) or combinations of these cellulose derivatives are used here as preferred materials.”).
Claim 24 is rejected under 35 U.S.C. 103 as being unpatentable over Bonnet, Ma, Hisaoka, further in view of Boerjan (US 6,149,956 A).
Regarding claim 24, Bonnet teaches wherein said at least one imaging system comprises a first imaging system (See the camera labeled in Fig. 1).
Bonnet fails to teach a second imaging system. However, Boerjan teaches a second imaging system being positioned one after the other along the conveyor system (Fig. 2 shows the scanning apparatus 106 for capturing images of the eggs. Fertilized eggs travel along the conveyor system 104 to the incubator 109 where a second scanning means 111 captures additional images of the eggs.).
Bonnet and Boerjan are analogous in the art to the claimed invention, because both teach observing a parameter eggs on a conveyor system for sorting. Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Bonnet’s invention by using a second imaging system downstream of the first imaging system. This modification allows for eggs to be further examined for fertility to further ensure that non-fertile eggs are not occupying space in the sorted eggs compartment, such as an incubator (Boerjan [Col. 5, lines 25-31] “…second scanning means 111 are arranged for obtaining NMR images of one or more eggs 107 in the incubator 109. These second scanning means 111 are arranged for determining whether a living embryo is present in the or each egg 107 in question, in order to determine whether the egg in question should be maintained in the incubator or should be removed from it.”).
Claim 29 is rejected under 35 U.S.C. 103 as being unpatentable over Bonnet, Ma, Hisaoka, further in view of Dirvanauskas et al. (Embryo development stage prediction algorithm for automated time lapse incubators. Computer Methods and Programs in Biomedicine. Volume 177. 2019.), hereafter Dirvanauskas.
Regarding claim 29, Bonnet fails to teach wherein the image analysis system employs an algorithm employs an algorithm built by a machine learning system. However, Dirvanauskas teaches wherein said image analysis system employs an algorithm built by a machine learning system utilising mathematical models based on sample or training images of eggs known to be fertilized (Fig. 1 shows images of different development stages of a fertilized egg cell. Table 2 shows results and each class, showing that the machine learning classifiers are capable of classifying images as having divided cells or not.).
Bonnet and Dirvanauskas are analogous in the art because both teach methods of examining cell division in the development stage of an embryo or egg. Therefore, it would have been obvious to one of ordinary skill in the art to modify Bonnet’s invention by utilizing machine learning to examine the plurality of images of each egg to determine if cell division in the blastodisc has occurred or not. This modification allows for machine learning to automate the process and save time over visual inspection by humans ([Section 1] “Embryologists select embryos by visual inspection, which requires time and is prone to error.”).
Conclusion
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure.
Kimmel et al. (Stages of embryonic development of the zebrafish. Dev. Dyn., 203: 253-310.) teaches observing the stages of fish eggs post fertilization using a dissecting stereo microscope.
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/Eric Shoemaker/
Patent Examiner
/JENNIFER MEHMOOD/Supervisory Patent Examiner, Art Unit 2664