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 .
Applicant’s submission filed on February 6, 2026 has been entered and considered. Rejections and/or objections not reiterated from the previous action mailed October 6, 2025 are hereby withdrawn. The following rejections and/or objections are either newly applied or are reiterated and are the only rejections and/or objections presently applied to the instant application. The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Election/Restrictions
Applicant’s election without traverse of Group 1, claims 1-15 drawn to a method to collect brain chads comprising extracting a brain region from a subject, slicing a 100-micron to 500-micron thick section on a cutting substrate comprising a gelling agent substrate and in a cutting solution; and coring the sliced section to form a brain chad; wherein the brain chad is employed in a subsequent cell and molecular biology/analysis and election of the species wherein the actuator is motorized (claim 3) in the reply filed on September 8, 2025 is acknowledged.
Applicant’s response dated September 8, 2025 indicates withdrawal of claims 4 and claims 15-20. Since claim 15 is encompassed by group 1 which has been elected for examination, this appears to be a typographical error in intending to refer to non-elected group 2, claims 16-20 and is interpreted as such for the purposes of examination. Claims 4 and 16-20 withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim.
Applicant has amended claims 1, 5, 8, 12, and 15. Claims 4 and 16-20 stand withdrawn.
Claims 1-3 and 5-15 are examined on the merits.
Priority
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. The instant application claims domestic benefit from U.S. provisional application 63/288117 filed on December 10, 2021.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on March 13, 2023 is in compliance with the provisions of 37 CFR 1.97 and has been considered by the examiner.
Objections to the Drawings
The drawings are objected to because:
FIG. 3 is objected to as failing to comply with 37 CFR 1.84(p)(5) because they do not include the following reference sign(s) mentioned in the description: Paras. [0020] and [0022] reference number 305 as indicating the sliced sample which is not present in FIG. 3. FIG 3 appears to use 105’ to indicate the sliced sample (also listed in Para. [0020] as the reference number for the sliced sample).
FIG. 4 is objected to as failing to comply with 37 CFR 1.84(p)(4) because reference characters "304’" and "404’" have both been used to designate the sample corer. See also Paras. [0025] and [0027] of the instant specification.
FIG. 4 is objected to as failing to comply with 37 CFR 1.84(p)(4) because reference characters "105’" (Para. [0024]), "405" (Paras. [0025], [0027]), and “105’’” (Paras. [0025], [0027]) are used to designate the sliced sample.
FIG. 4 is objected to as failing to comply with 37 CFR 1.84(p)(5) because they do not include the following reference sign(s) mentioned in the description: Para. [0024] references number 303 as indicating a sample chad. Number 303 is not present in FIG. 4.
FIGs. 5 and 6 are objected to as failing to comply with 37 CFR 1.84(p)(5) because they do not include the following reference sign(s) mentioned in the description: Para. [0029] references number 303 as indicating a sample chad. Number 303 is not present in FIGs. 5 or 6.
FIGs 5 and 6 are objected to as failing to comply with 37 CFR 1.84(p)(4) because reference characters "402’" and "402’’" have both been used to designate the exoscope. See also Paras. [0029], [0030], and [0036] of the instant specification.
This is not intended to be a comprehensive list. It is recommended that Applicant carefully review the specification for other instances of discrepancies between the specification and the drawings.
Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
New Claim Rejections - 35 USC § 112(a)
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.
This is a new rejection necessitated by Applicant’s amendment.
Claims 1-3 and 5-15 are 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.
Claim 1, amended in the response filed February 6, 2026 recites “a method to collect and analyze brain chads consisting of neuronal assemblies having multiple layers of cells organized as a neural circuit…to evaluate neuro modulators, neurotransmitters, and proteins at synapses and receptors…” and “wherein each tissue punch of tissue collected under visual guidance comprises one or more neuronal assemblies…”. Claim 1 recites “coring the sliced 100 micron to 500 micron thick section to form a brain chad while guided via a wide-angle field scope…”. Claim also 1 recites “molecular analysis on cellular and molecular changes across the brain region of neuronal assemblies…”.
Dependent claim 5, also amended in the response filed February 6, 2026 recites “a visual guide wide-angle-field-of-view exoscope…”.
Applicant discloses in their remarks that support for the amended claims is to be found in Paras. [0005]-[0006] and [0040]-[0051] of the instant specification. Para. [0006] discloses a singular mention of brain chads which comprise cells organized as “a neural circuit”, but does not disclose “neuronal assemblies” or “one or more neuronal assemblies”. Para. [0041] discloses that the brain chads contain synapses and receptors and can allow for sampling of neuromodulators, neurotransmitters, and proteins, however there is no disclosure of measurement of neuromodulators, neurotransmitters, and proteins which are found at synapses and receptors. For example, Applicant’s instant specification does not indicate an ability to detect global presence of neuromodulators in a tissue punch from neuromodulators which are specifically present at a synapse or at a receptor. Therefore, clear support for the new limitations of regarding neuronal assemblies and the ability to precisely measure neuromodulators, neurotransmitters, and proteins found at the synapse and/or receptor cannot be found in the instant application. Further, para. [0004] of the instant specification discloses cellular and molecular changes across “juxtaposed brain regions”, there is no support for the ability to analyze cellular and molecular changes across a single “brain region of neuronal assemblies”, as instantly claimed. Since Applicant’s method involves analysis of the entire brain chad, there is no indication of the ability to analyze changes across a single brain chad.
Similarly, while applicant has support for use of an exoscope and claim 5 prior to the current amendment recited “a visual guide having a wide field view” there is no support found in instant specification regarding use of a “wide-angle field scope” as recited in amended claim 1 or “a visual guide wide-angle-field-of-view exoscope” as recited in amended claim 5. Accordingly, the amendments to claims 1 and 5 are considered to constitute new matter.
MPEP 2163.06 notes “If new matter is added to the claims, the examiner should reject the claims under 35 U.S.C. 112, first paragraph - written description requirement. In re Rasmussen, 650 F.2d 1212, 211 USPQ 323 (CCPA 1981).” MPEP 2163.02 teaches that “Whenever the issue arises, the fundamental factual inquiry is whether a claim defines an invention that is clearly conveyed to those skilled in the art at the time the application was filed...If a claim is amended to include subject matter, limitations, or terminology not present in the application as filed, involving a departure from, addition to, or deletion from the disclosure of the application as filed, the examiner should conclude that the claimed subject matter is not described in that application”. MPEP 2163.06 further notes “When an amendment is filed in reply to an objection or rejection based on 35 U.S.C. 112, first paragraph, a study of the entire application is often necessary to determine whether or not “new matter” is involved. Applicant should therefore specifically point out the support for any amendments made to the disclosure”.
Applicant has not specifically pointed out support in the originally filed application for the amended claims in the remarks filed on February 6, 2026. If Applicant believes that support for the new limitations, as now recited in Claims 1 and 5 is present and clearly envisaged in the instant application or earlier filed priority documents, applicant must, in responding to this Office Action, point out with particularity, where such support may be found. Declarations and new references cannot demonstrate possession of a concept after the fact.
Dependent claims 2-3 and 5-15 have been included in the rejection base on their dependency on a claim 1.
New Claim Rejections - 35 USC § 112(b)
Claims 1, 8 and 14-15 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 1 recites that brain chads are formed by “coring” of a 100-micron to 500-micron thick section and also “wherein the brain chad includes tissue punches and tissues from acute brain slices within the sliced 100-micron to 500-micron thick section” which renders the scope of the claim indefinite. As claimed, it is not clear if the brain chads are cored, which would indicate that the brain chads are a tissue punch or if the brain chads are tissues from thinner sub-slices from the 100-micron to 500-micron thick section or both. Appropriate correction is required.
Claim 1 recites “the brain region of neuronal assemblies” in lines 17-18. There is insufficient antecedent basis for this limitation in the claim as the prior recitation is to “a brain region” which is then cored to form a “chad” consisting of neuronal assemblies. Appropriate correction is required.
Claim 8 which depends from claim 1, recites the limitation "the subsequent area-specific brain cell and molecular analysis" in lines 1-2. There is insufficient antecedent basis for this limitation in the claim as claim 1 recites “area-specific brain cell and molecular analysis”. Appropriate correction is required.
Claims 14 and 15, which depend from claim 1, recite the limitation "the subsequent cell and molecular analysis" in lines 1-2. There is insufficient antecedent basis for this limitation in the claim as claim 1 has been amended to recite “area-specific brain cell and molecular analysis”. Appropriate correction is required.
New Claim Rejections - 35 USC § 103
This is a new rejection necessitated by Applicant’s amendment. However, this rejection shares some similarity to the rejection as previously set forth in the office action dated October 6, 2025. Any aspect of Applicant’s traversal that pertains to the rejection as newly set forth will be provided following the new statement of rejection.
Applicant has amended claim 1 to recite “multiple layers of cells organized as a neural circuit”. The broadest reasonable interpretation of neural circuit is, at minimum, two cells which are connected at a synapse. As Applicant’s claims are drawn to collection of tissue punches from brain slices (i.e., the Palkovits micropunch technique), which is well-known and commonly used by those having ordinary skill in the art, and whole tissue punch analysis, all tissue punches would be generally understood by a skilled artisan to comprise multiple layers of cells and have at least two cells which are organized as a neural circuit. Similarly, since all tissue punches retain the structure of the area from which they were obtained, all tissue punches are considered to be “area specific” and would be generally be understood by a skilled artisan to contain neuromodulators, neurotransmitters, proteins, etc. which were present in the area at the time that the “area specific” tissue punch was obtained.
Claims 1-3, 6-12, and 14-15 are rejected under 35 U.S.C. 103 as being unpatentable over Merrill et al. (2017, Patch clamp-assisted single neuron lipidomics. Scientific Reports, 7(1), 5318, hereafter “Merrill” in view of Jacobowitz and Kallarakal (2004, Flotillin-1 in the substantia nigra of the Parkinson brain and a predominant localization in catecholaminergic nerves in the rat brain. Neurotoxicity Res., 6(4), 245-257) and Johnson et al. (2018, Vital ex vivo tissue labeling and pathology-guided micropunching to characterize cellular heterogeneity in the tissue microenvironment. Biotechniques, 64(1), 13-19, hereafter “Johnson”).
With regard to claim 1, Merrill teaches a method of obtaining brain tissue for use in patch clamp, lipid, and protein analysis which comprises obtaining tissue punches, which are considered to read on brain chads, from brain slices. Merrill teaches a method of obtaining brain chads wherein brains are extracted; sectioned into 400 µm thick slices to isolate brain areas containing cells of interest (e.g., dentate gyrus); placed on a submerged net, which is considered to read on a cutting substrate; incubated in artificial cerebrospinal fluid, which is considered to read on a cutting solution (Materials and Methods, Slice preparation); and cylinders of brain tissue (i.e., brain tissue punches) were collected using a manual punch to be used in lipid analysis and protein extraction and quantification, which is considered to read on analyzing brain chads to evaluate neuromodulators and proteins (Materials and Methods, tissue punch Collection, lines 5-7). Merrill teaches that punches housed in vials were processed for lipid analysis using the same method as for single neurons (Pg. 6, Tissue punch collection) which comprises centrifugation of the brain chad and subjecting the output of the centrifugation to a biological assay (i.e., liquid chromatography/mass spectrometry (LC-MS)) (Pg. 7, Lipid extraction from single neurons).
Although Merrill teaches subjecting the brain chad to centrifugation and subjecting the output of that centrifugation to a biological assay. Merrill is silent as to use of any of the instantly claimed biological assays.
Merrill teaches that accumulation of certain lipids (i.e., sphingomyelin) could be indicative of expansion of lipid rafts and accelerated trafficking of glutamate receptors (Pg. 6, 1st para.).
Jacobowitz and Kallarakal teach a study of gene expression in Parkinson’s disease which identified flotillin-1, which is a membrane protein found in lipid rafts as a gene/protein of interest upregulated in certain areas of the brain in Parkinson’s disease (Abstract). Jacobowitz and Kallarakal further teach that lipid rafts and flotillin-1 are associated with Alzheimer’s disease (Pg. 255, right col, 3rd full para.). Jacobowitz and Kallarakal teach use of a micropunch procedure to isolate dopamine containing cells in human brains from approximately 120 micron sections (Pg. 246, left col., middle para.), and to collect samples from various areas within the rat brain (including substantia nigra pars compacta, supraoptic nucleus, locus coeruleus, hippocampus, cortex, and cerebellum) (Pg. 247, right col, last para.); which is considered to reasonably read on collection of brain chads. Jacobowitz and Kallarakal teach that the collected micropunches from rat brain were used for a western blot (i.e., immunoblotting) for flotillin-1 (Pg. 247, right col, last para.).
Therefore, it would have been obvious to one having ordinary skill in the art, before the effective filing date of the claimed invention, to use an immunoblotting assay for analysis of brain chads isolated various parts of the brain as taught by Jacobowitz and Kallarakal in the method of collecting and analyzing brain chads for lipids as taught by Merrill with a reasonable expectation of success. A skilled artisan would have been motivated to do so in order to evaluate localized expression of lipid-associated proteins such as flotillin-1 which are known to be associated with neurological diseases such as Alzheimer’s and Parkinson’s diseases. As Merrill teaches analysis of lipids in localized brain areas and that lipid homeostatsis is known to play a role in neurodegenerative diseases such as Alzheimer’s and Parkinson’s diseases, a skilled artisan would have recognized that immunoblotting could be used to assay lipid associated proteins such as flotillin-1 which are altered in the same neurological diseases. One having ordinary skill in the art would have had a reasonable expectation of success as both Merrill and Jacobowitz and Kallarakal teach use of collection of brain chads from certain brain areas and subsequent localized analysis of those chads using biological assays.
Additionally, it is noted that, the brain tissue punch technique is considered routine and conventional and cellular and molecular analysis of brain tissue punches would be widely known to those having ordinary skill in the art. Additionally, immunoblotting, ELISA, genotyping, cloning, and PCR techniques are commonly used cellular/molecular assays and would also be widely known to those having ordinary skill in the art.
Merrill is silent as to the use of a cutting substrate comprising a gelling agent. However, cutting substrates comprising agar are widely used and well known to those having ordinary skill in the art. Use of a cutting substrate comprising agar is considered to reasonably read on a cutting agent comprising a gelling substrate.
While Merrill and Jacobowitz and Kallarakal both teach collection of brain chads from various brain areas, both are silent as to the specific visual guide. However, one of ordinary skill would be aware that these techniques are generally performed under visual guide from a microscope due to the small sizes of the brain chads being collected - less than 1 mm in Merrill (Pg. 6, Tissue punch collection) and 1 mm in Jacobowitz and Kallarakal (Pg. 246, left col., middle para.).
Johnson teaches a method of obtaining tissue micropunches from animal tissue slices comprising slicing approximately 200 micron thick sections (Pg. 14, left col., last para.) and subsequent micropunching of tissue which is performed using a 2x objective and an inverted microscope (Pg. 14, right col., last para.), which is considered to reasonably read on coring of sections to form brain chads while guided via a wide angle field scope.
Therefore it would have been obvious, to one having ordinary skill in the art, before the effective filing date of the claimed invention, to use a wide angle field scope as taught by Johnson in the method of collecting and analyzing brain chads as taught by Merrill and Jacobowitz and Kallarakal with a reasonable expectation of success. A skilled artisan would have been motivated to use a wide angle field scope as taught by Johnson because Johnson teaches that this is a successful technique for being able to isolate micropunches from sliced tissue sections. One of ordinary skill in the art would have had a reasonable expectation of success as Merrill, Jacobowitz and Kallarakal, and Johnson all teach collection of tissue micropunches from tissue slices.
With regard to claims 2 and 3, the combination of Merrill and Jacobowitz and Kallarakal discloses the claimed invention except Merrill and Jacobowitz and Kallarakal teach use of a manual punch. It would have been obvious to one having ordinary skill in the art at the time the invention was made to replace a manual punch with a motorized mechanically guided actuator, since it has been held that broadly providing a mechanical or automatic means to replace manual activity which has accomplished the same result involves only routine skill in the art. In re Venner, 120 USPQ 192.
With regard to claim 6, Merrill teaches wherein brains were sectioned into 400 µm thick sections, placed on a submerged net (i.e., a cutting substrate), and incubated in artificial cerebrospinal fluid (Materials and Methods, Slice preparation, lines 4-7).
With regard to claim 7, as detailed above, gelling agent substrates comprising agar are widely known to those having ordinary skill in the art. A skilled artisan would be aware that agar is a powder that must be mixed with a liquid. As Merrill teaches that the cutting substrate is aCSF, it would have been obvious to a skilled artisan to mix of agar with aCSF.
With regard to claims 8 and 14-15, Applicant has amended claims 8 and 14-15 to recite “localized changes in cellular and molecular parameters…”. As detailed above in claim 1, all tissue punches are interpreted to be “area specific” and therefore the recitation of “localized” is interpreted to refer to mean local to the area of the brain chad. Therefore, as claim 1 recites analysis of the entirety of the collected brain chad, all analysis is interpreted to be indicative of localized changes.
As detailed above in claim 1, Merrill teaches that collected brain chads where collected from the dentate gyrus and the hippocampus proper (Materials and Methods, Tissue punch collection, line 6), which is considered to read on juxtaposed brain regions, followed by subsequent lipid analysis using biological assays (liquid chromatography/mass spectrometry) and protein quantification using a BCA assay of the brain chads (Materials and Methods, Tissue punch collection, lines 8-9) which is considered read on providing information regarding localized changes. Additionally, Merrill teaches that disruptions of lipid homeostasis is known to contribute to neurological diseases such as Alzheimer’s disease and Parkinson’s disease (Pg. 1, 2nd Para., lines 2-4) and Merrill further indicated that their methods are applicable to other studies in healthy and diseased brains (Pg. 6, 2nd para). Jacobowitz and Kallarakal teach altered expression of flotillin-1, which is a membrane protein found in lipid rafts, in certain areas of the brain in Parkinson’s disease (Abstract). Jacobowitz and Kallarakal further teach that lipid rafts and flotillin-1 are also associated with Alzheimer’s disease (Pg. 255, right col, 3rd full para.). Jacobowitz and Kallarakal teach collection of brain chads from various areas in the rat brain in order to perform biological assays to analyze area specific differences in flotillin-1 expression. Therefore, the combined teachings of Merrill and Jacobowitz and Kallarakal are considered to provide support for area specific analysis of differences in cellular and molecular parameters which are associated with neurological diseases such as Alzheimer’s and Parkinson’s diseases.
With regard to claim 9, Merrill teaches wherein brain chad samples were homogenized in a buffer prior to protein quantification (Materials and Methods, Tissue punch collection, lines 11-12). A skilled artisan would recognize that tissue homogenization would cause cell lysis and cellular membrane disruption and result in protein extraction.
With regard to claim 11, Merrill teaches wherein protein quantification of brain chads was performed using a Pierce BCA kit (Materials and Methods, Tissue punch collection, line 10).
With regard to claim 12, Merrill teaches wherein brain slices were placed on a cutting substrate, incubated with artificial cerebrospinal fluid “containing (in mM) containing (in mM) 119 NaCl, 26 NaHCO3, 2.5 KCl, 1.0 NaH2PO4, 2.5 CaCl2, 2.7 MgSO4, and 11 glucose, saturated with 95% O2/5% CO2 (pH 7.4)” (Materials and Methods, Slice preparation, lines 7-8) and further that brain slices were transferred to a chamber and continuously perfused with oxygenated artificial cerebrospinal fluid (Materials and Methods, Tissue punch collection, lines 2-3). This is considered to reasonably read on storing the brain region “alive and equilibrated”.
Claims 2-3 and 5 are rejected under 35 U.S.C. 103 as being unpatentable over Merrill, Jacobowitz and Kallarakal, and Johnson as applied to claim 1 above and further in view of Zhao et al. (CN113358432A, Espacenet translation attached, hereafter “Zhao”)
With regard to claims 2-3 and 5, as detailed above, Merrill teaches a method of obtaining brain tissue punches and subsequent analysis of proteins and lipids of the collected “brain chads”. Jacobowitz and Kallarakal teach a study of lipid associated proteins comprising collection of brain chads and subsequent immunoblotting via Western blot. Johnson teaches a method of obtaining tissue micropunches from animal tissue slices where the punches are collected under visual guidance using a wide angle scope.
The combination of Merrill, Jacobowitz and Kallarakal, and Johnson is silent as to the use of a motorized mechanically guided actuator or coring of the section via an exoscope.
Zhao teaches a device for micro region sampling from tissue sections (Para. [0004]), which can be fresh tissue sections (Para. [0011]), comprising an inverted microscope imaging module (Para. [0005]) and a microneedle positioned above the tissue section (Para. [0006]). Zhao teaches that movement of the platform holding the tissue sample and the micromanipulation system are controlled by software (Para. [0010], [0021]). Zhao teaches that the device scans the tissue section to select a region of interest, the site is moved below the microneedle, and the microneedle is lowered using an electric manipulation system (Para. [0025]), which is considered to reasonably read on coring via mechanically guided actuator which is motorized. Zhao teaches that this system provides the benefit of obtaining location information of the microregion and can obtain high quality RNA, DNA, and proteins for downstream analysis (Para. [0015]). Further, Zhao teaches that the automated sampling system is equipped with multiple imaging systems for tissue sample imaging laterally as well as below, these systems are connected to imaging sensors see (Para. [0014]), and that these systems are connected to a computer which allows for conversion of images into digital information and subsequent display of the collected images via software (Paras. [0021] and [0023]), which is considered to reasonably read on an exoscope. Zhao teaches that this system allows from real time monitoring of the tissue collection (Paras. [0023], [0025]).
Therefore, it would have been obvious to one having ordinary skill in the art, before the effective filing date of the claimed invention to use the automated sampling system configured as an exoscope as taught by Zhao in the method of collecting and analyzing brain chads as taught by the combination of Merrill, Jacobowitz and Kallarakal, and Johnson with a reasonable expectation of success. A skilled artisan would have been motived to use the automated system as taught by Zhao because Zhao teaches that the system can retain positional information related to the area of sample collection and also obtain high quality RNA, DNA, and proteins for downstream analysis. One having ordinary skill in the art would have had a reasonable expectation of success because Merrill, Jacobowitz and Kallarakal, Johnson, and Zhao all teach collection of micrometer sized samples from tissue sections.
Claim 13 is rejected under 35 U.S.C. 103 as being unpatentable over Merrill, Jacobowitz and Kallarakal, and Johnson, as applied to claim 1 above and further in view of Ziv and Schiller (2007, Differential interference contrast (DIC) imaging of living cells. Cold Spring Harbor Protocols, 2007(7), pdb-prot4787).
With regard to claim 13, as detailed above in claim 1, Merrill teaches a method of obtaining brain tissue punches and subsequent analysis of proteins and lipids of the collected “brain chads”. Jacobowitz and Kallarakal teach a study of lipid associated proteins comprising collection of brain chads and subsequent immunoblotting via Western blot. Johnson teaches a method of obtaining tissue micropunches from animal tissue slices where the punches are collected under visual guidance using a wide angle scope.
The combination of Merrill, Jacobowitz and Kallarakal, and Johnson is silent as to the use of differential interference contrast for coring of the brain chad.
Ziv and Schiller teach that use of differential interference contrast (DIC) is a useful technique for imaging of living cells and can be applied for use in brain slices. Ziv and Schiller teach that DIC can be used with inverted microscopes and is valuable for complementing fluorescent images (Introduction).
Therefore, it would have been obvious to one having ordinary skill in the art, before the effective filing date of the claimed invention, apply the technique of coring brain chads under DIC as taught by Ziv and Schiller to the method of collecting and analyzing brain chads where brain chads are cored under visual guidance via a wide angle field scope as taught by the combination of Merrill, Jacobowitz and Kallarakal, and Johnson with a reasonable expectation of success. A skilled artisan would have been motivated to combine coring under DIC as taught by Ziv and Schiller with coring under a wide angle field scope because Johnson’s coring method uses fluorescent images and Ziv and Schiller teach that use of DIC imaging provides a valuable complement to fluorescent microscopy data. One having ordinary skill in the art would have had a reasonable expectation of success as both Johnson and Ziv and Schiller teach microscopy techniques useful for imaging of tissue slices using an inverted microscope.
Response to Arguments
Applicant's arguments filed February 6, 2026 have been fully considered but they are not persuasive.
Applicant traverses on Pg. 10 that Merrill and Stachniak do not teach or disclose all the features of amended claim 1, specifically “coring the 100 micron to 500 micron thick section to form a brain chad while guided via a wide-angle field scope configured to visually guide harvesting of sample chads wherein the brain chad includes tissue punches and tissues from acute brain slices within the sliced 100 micron to 500 micron thick section, wherein each tissue punch or tissue collected under visual guidance comprises one or more neuronal assemblies having (i) multiple layers of cells organized as a neural circuit and (ii) neuromodulators, neurotransmitters, and proteins at synapses and receptors in the neural circuit” and “performing area-specific brain cell and molecular analysis…by subjecting the entirety of the brain chad to centrifugation and subjecting an output of the centrifugation to (i) a biological assay selected from the group consisting of immune blotting…ELISA, and MEKC or (ii) a biological assay selected from the group consisting of genotyping, cloning, and qPCR”. Applicant asserts that none of the cited references cure the deficiencies of Merrill and Stachniak as it relates to amended claim 1.
Applicant asserts that Merrill discloses a method for single-neuron lipidomics including collecting a tissue punch from a brain slice and is silent as to performing direct analysis of the entirety of the tissue punch. Applicant asserts that Merrill states a “punch was taken from the hippocampus proper for protein quantification” and “Lipids were extracted, separated, and analyzed in the same manner as single-neuron samples” (Pg. 10, 2nd para.).
Applicant asserts that Stachniak discloses a whole cell analysis performed on cell culture which comprises collection of tissue punches which are transferred to a culture dish with dorso-ventral orientation maintained and for slice cultures, brains were bisected in the horizontal plane and the ventral half was placed on a block of agar ventral side up before manual sectioning.
Applicant asserts on Pg. 11 that none of the other references cure the deficiencies of Merrill and Stachniak as Yoda is directed to collection of multiple tissue microdissections while maintaining positional information using frozen brain sections and Badnish is directed to electrophysiology of brain tissue punches.
Applicant’s arguments have been fully considered but are not persuasive.
With regard to Applicant’s assertion that Merrill is “silent as to performing direct analysis on the entirety of the tissue punch”, Merrill does teach collection of punches from areas of the brain for protein quantification (as Applicant has admitted on Pg. 10) as well as lipid analysis. Applicant’s arguments with respect the prior art of Stachniak, Yoda, and Badnish are moot based on the new ground of rejection.
Conclusion
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 ERIN V PAULUS whose telephone number is (571)272-6301. The examiner can normally be reached Mon-Fri 8 AM-5 PM.
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/ERIN V PAULUS/Examiner, Art Unit 1631
/ARTHUR S LEONARD/Examiner, Art Unit 1631
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