DETAILED ACTION
The amendment filed on 09/17/2026 has been entered and fully considered. Claims 1-20 are pending, of which claim 1, 4, 11 and 13 are amended.
Response to Amendment
In response to amendment, the examiner maintains rejection over the prior art established in the previous Office 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 .
Claim Rejections - 35 USC § 103
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claim(s) 1-3 and 11-12 is/are rejected under 35 U.S.C. 103 as being unpatentable over Shikimura et al. (JP 2019-213464, IDS) (Shikimura) in view of Hanson (US 2013/0210063).
Regarding claim 1, Shikimura discloses a measurement unit used for measuring a characteristic of a biological sample (par [0010]), the measurement unit comprising:
a first container that includes a first housing space (par [0049]), a first opening communicating with the first housing space (par [0050), and a light transmission region through which light generated in the first housing space is transmitted (par [0004]); and
a first solution that is a mixture of a plurality of reagents containing an indicator reacting with a component generated from the biological sample (par [0049][0026]), the indicator being an indicator reacting with hypochlorous acid or a halogen equivalent thereof produced by myeloperoxidase (par [0026] [0027]), or an indicator reacting with superoxide (par [0028]).
Shikimura does not specifically disclose wherein the first solution is housed in the first housing space in a state of being frozen.
However, Hanson teaches a pre-made, ready-to-use indicator-containing reagent solution that may be stored frozen. Hanson states:
“It is preferred that the reagent solution be present in the form of a pre-made, ready to use reagent solution.” (par [0072]).
Hanson further states:
“The preferred reagent solution is stable at room temperature (e.g., about 25° C.), refrigerated (e.g., about 4° C.) or frozen (e.g., about −20° C.).” (par [0072]).
Hanson also expressly teaches that the pre-made reagent solution is housed in a container:
“In some embodiments, the kit includes a reagent solution comprising resazurin, a phosphate buffer and an aqueous solvent . . . disposed in a container.” (par [0073][0024]).
Hanson’s reagent solution is a mixture of reagents including resazurin, buffer, and aqueous solvent, and resazurin is an indicator that is reduced by the metabolic activity of viable cells to produce fluorescent resorufin (par [0006], [0043], [0065]).
Shikimura and Hanson are analogous art because they are from the same field of endeavor and contain functional similarities. Both relate to optical measurement of biological-cell activity using a premixed, indicator-containing reagent solution and detection of fluorescence generated through interaction between the indicator and the biological cells.
It would have been obvious to one of ordinary skill in the art before the effective filing date to modify Shikimura’s measurement unit by preloading Shikimura’s mixture of APF, MCLA, CaCl₂, and buffer into the housing space of Shikimura’s sample container and storing the preloaded solution in a frozen state, as taught by Hanson. Hanson expressly identifies a pre-made, ready-to-use reagent solution disposed in a container and expressly identifies frozen storage at approximately −20° C. as a suitable storage condition. One of ordinary skill in the art would have been motivated to preserve the premixed indicator reagent in a stable, ready-to-use condition until the biological assay is performed, as suggested by Hanson, while reducing the reagent preparation and transfer required at the time of measurement. The modification would have involved the predictable use of Hanson’s disclosed frozen-storage option for the premixed reagent solution in Shikimura’s known optical biological-sample measurement container, with a reasonable expectation that the solution could be thawed and used for its intended optical assay.
Regarding claim 2, Shikimura teaches that wherein the biological sample contains a white blood cell (neutrophil cells) (par [0022]).
Regarding claim 3, Shikimura discloses that wherein the indicator is a fluorescent indicator (par [0026]).
Regarding claim 11, Shikimura teaches a method for manufacturing a measurement unit used for measuring a characteristic of a biological sample, the method comprising:
a first preparation step of preparing a first container that includes a first housing space, a first opening communicating with the first housing space (Fig. 1, par [0049]), and a light transmission region through which light generated in the first housing space is transmitted (par [0004]); wherein
the indicator is an indicator reacting with hypochlorous acid or a halogen equivalent thereof produced by myeloperoxidase (par [0026] [0027]), or an indicator reacting with superoxide (par [0028]).
Shikimura does not specifically teach a first freezing step of freezing a first solution that is a mixture of a plurality of reagents containing an indicator reacting with a component generated from the biological sample in a state of being housed in the first housing space.
Hanson teaches that an indicator-containing reagent solution is preferably provided as:
“a pre-made, ready to use reagent solution” and
that the reagent solution is stable when:
“frozen (e.g., about −20° C.).” (par [0072])
Hanson further teaches a kit containing a reagent solution comprising resazurin, phosphate buffer, and aqueous solvent “disposed in a container.” (par [0073]).
Shikimura and Hanson are analogous art because both relate to optically measuring biological-cell activity using a premixed indicator-containing reagent solution. It would have been obvious to one of ordinary skill in the art to manufacture Shikimura’s measurement unit by introducing Shikimura’s premixed indicator solution into the housing space of the sample container and freezing the solution in that container, as suggested by Hanson’s frozen, ready-to-use reagent solution disposed in a container. One of ordinary skill in the art would have been motivated to maintain the premixed indicator solution in a stable, ready-to-use condition until the assay is performed, as expressly suggested by Hanson.
Regarding claim 12, Shikimura in view of Hanson teaches:
a solution production step of producing the first solution, wherein in the solution production step, the plurality of reagents are mixed outside the first housing space.
Shikimura teaches producing the solution by mixing MCLA, APF, CaCl₂, HEPES, NaCl, and KCl before the solution is combined with the biological sample (para. [0049]).
Hanson additionally teaches that the reagent solution is “pre-made” and subsequently “disposed in a container.” (par [0072]–[0073]). Thus, Hanson teaches producing the completed reagent solution before placing it in the container, rather than separately adding the individual reagents to the measurement container.
It would have been obvious to mix Shikimura’s plurality of reagents outside the sample-container housing space and then dispense the completed solution into the container. This is the predictable manner of implementing Hanson’s pre-made, ready-to-use reagent solution and permits the same premixed solution to be loaded into one or more measurement containers before frozen storage.
Claim 4 and 13 is rejected under 35 U.S.C. § 103 as being unpatentable over Shikimura in view of Hanson, and further in view of Nakata et al. (US 2015/0219548) (Nakata).
Regarding claim 4, Shikimura and Hanson do not expressly teach:
a rotor that is housed in the first housing space to be immersed in the first solution.
However, Nakata teaches this additional limitation. In particular, Nakata discloses:
“Reference numeral 7 denotes a rotor that allows the sample solution S contained in the sample container 5. The rotor 7 is housed in the sample container 5 along with the sample solution S and an emission reagent, and is rotationally driven by a magnetic stirrer 27 incorporated in the measurement section 6 when the sample container 5 is housed in the measurement section 6.” (par [0063]).
Nakata’s rotor 7 is therefore housed in the internal space of sample container 5 together with, and immersed in, the liquid sample solution and emission reagent. Nakata further explains that the rotor permits the sample solution and emission reagent to be stirred and mixed at a predetermined temperature, thereby permitting microorganisms to emit light rapidly and enabling the optical measurement to be performed quickly (par [0063]).
Shikimura, Hanson, and Nakata are analogous art because they relate to optically measuring a biological characteristic using a sample container containing a biological sample and an indicator-containing reagent solution. Shikimura measures fluorescence and chemiluminescence from a biological sample containing blood and indicator reagents; Hanson teaches storing an indicator-containing reagent solution in a frozen, ready-to-use state; and Nakata teaches placing a rotor within an optically measured sample container to stir and mix the biological sample and emission reagent.
It would have been obvious to one of ordinary skill in the art to provide Nakata’s rotor within the housing space of Shikimura’s sample container so that the rotor is immersed in Shikimura’s indicator-containing reagent solution. One of ordinary skill in the art would have been motivated to promote mixing of the biological sample with the indicator-containing reagent solution and thereby facilitate rapid and consistent optical measurement, as suggested by Nakata. Because the Shikimura-Hanson combination stores the reagent solution frozen in the sample container, the rotor preloaded within that container would remain housed in the housing space and immersed in the solution when the solution is frozen, producing the measurement unit recited in claim 4. The modification would have been a predictable application of Nakata’s known internal rotor to the optical biological-sample measurement container of Shikimura and Hanson, with a reasonable expectation of successfully mixing the solution after thawing.
Nakata teaches the functional arrangement: a rotor inside the container, immersed in the solution, used for stirring/mixing. It would have been obvious to modify Nakata's container to include a protruding support (such as a bottom protrusion or bearing projection) to physically support the rotor. Such supports are conventional in magnetic stirring vessels to reduce friction, maintain rotor position, and improve stable rotation.
Regarding claim 13, Regarding:
in the first freezing step, the first solution is frozen in a state where a rotor housed in the first housing space is immersed in the first solution,
Nakata teaches that:
“The rotor 7 is housed in the sample container 5 along with the sample solution S and an emission reagent.” (par [0063]).
Nakata’s rotor is therefore housed within and immersed in the indicator-containing liquid in the sample container. Nakata further teaches rotating the rotor using a magnetic stirrer so that the sample and emission reagent are stirred and mixed at a predetermined temperature, permitting rapid optical measurement (par [0063]).
Shikimura, Hanson, and Nakata are analogous art because they relate to optical measurement of biological samples using indicator-containing reagent solutions housed in sample containers. It would have been obvious to place Nakata’s rotor into Shikimura’s sample container before introducing and freezing the indicator-containing solution. The rotor would consequently remain immersed in the solution during the freezing step. One of ordinary skill in the art would have been motivated to provide the rotor so that, after thawing, the biological sample and indicator-containing solution could be rapidly and uniformly mixed, as suggested by Nakata.
Nakata teaches the functional arrangement: a rotor inside the container, immersed in the solution, used for stirring/mixing. It would have been obvious to modify Nakata's container to include a protruding support (such as a bottom protrusion or bearing projection) to physically support the rotor. Such supports are conventional in magnetic stirring vessels to reduce friction, maintain rotor position, and improve stable rotation.
Claims 5-6 are rejected under 35 U.S.C. § 103 as being unpatentable over Shikimura in view of Hanson, and further in view of Wohlstadter et al. (US 2004/0022677 A1).
Regarding claim 5, Shikimura in view of Hanson does not expressly teach:
“the first container has a flat plate shape.”
Wohlstadter teaches an assay container having a flat plate shape, stating:
“Preferred embodiments of the invention are multi-well assay plates that use industry standard multi-well plate formats for the number, size, shape and configuration of the plate and wells,” and “[t]he assay plates are preferably flat, but may also be curved (not flat).” (par [0086] [0087]).
Wohlstadter further teaches that the plates are suitable for fluorescence, chemiluminescence, bioluminescence, and other optically measured assays (par [0015] [0016] [0096])
Shikimura, Hanson, and Wohlstadter are analogous art because they relate to optical measurement of biological samples using indicator-containing reagents housed in assay containers. It would have been obvious to one of ordinary skill in the art to configure the sample container of the Shikimura-Hanson measurement unit as Wohlstadter’s flat assay plate. One of ordinary skill in the art would have been motivated to improve compatibility with conventional optical-assay equipment and facilitate efficient handling and optical measurement of biological samples, as suggested by Wohlstadter’s use of industry-standard flat assay plates. Therefore, the combination teaches the limitation of claim 5.
Regarding claim 6, Shikimura in view of Hanson does not expressly teach:
“the first container includes an inner surface having a curved surface shape.”
Wohlstadter teaches a well forming the sample-housing space of an assay plate and having a curved inner surface:
“Each of wells 158, preferably, comprise a wall 162, an interior surface 164, a counter electrode 166, and a working electrode 168. As shown, wall 162 may define a cylindrical volume.” (par [0138]).
Wohlstadter further states:
“Wall 162 has an interior surface 164 that is preferably cylindrical in shape and defines a volume of well 158.” (par [0139]).
Wohlstadter explains that the cylindrical interior surface, together with the bottom surfaces, forms a container suitable for holding liquids (par [0138] [0139]).
It would have been obvious to one of ordinary skill in the art to provide the sample-housing space of the Shikimura-Hanson measurement unit with Wohlstadter’s cylindrical inner surface. One of ordinary skill in the art would have been motivated to improve liquid containment and provide a conventional well geometry suitable for receiving, mixing, and optically measuring the biological sample and indicator-containing reagent solution, as suggested by Wohlstadter.
Claims 7-8, 10, 14-16 and 18-19 are rejected under 35 U.S.C. § 103 as being unpatentable over Shikimura in view of Hanson, and further in view of Cathcart et al. (US 5,443,791) (Cathcart).
Regarding claim 7, Regarding “a second container that includes a second housing space and a second opening communicating with the second housing space,”
Cathcart teaches a pipette tip configured to receive and subsequently dispense an aspirated liquid. Cathcart states:
“After the liquid is aspirated, the probe tip is slowly withdrawn…. The probe is then moved to wherever is required to dispense the liquid that has been aspirated.” (col. 21, line 14-22).
Cathcart’s pipette tip necessarily includes an internal space in which the aspirated liquid is held and a tip opening communicating with that internal space through which the liquid is aspirated and dispensed. Thus, the internal space of Cathcart’s pipette tip corresponds to the second housing space, and the distal opening corresponds to the second opening.
Regarding “a second solution that contains a stimulant for activating a function of the biological sample,” Shikimura teaches:
“After 2.5 minutes, a protein kinase C activator (PMA (phorbol 12-myristate 13-acetate): neutrophil stimulator) was added to the biological sample to a concentration of 0.1 μM.” (par [0050]).
PMA is therefore a stimulant that activates a function of the biological sample, namely neutrophil myeloperoxidase activity and superoxide-production activity.
Shikimura and Cathcart do not expressly teach “wherein the second solution is housed in the second housing space in a state of being frozen.”
Hanson teaches that a premade biological-assay reagent solution may be stored frozen:
“It is preferred that the reagent solution be present in the form of a pre-made, ready to use reagent solution…. The preferred reagent solution is stable at room temperature (e.g., about 25° C.), refrigerated (e.g., about 4° C.) or frozen (e.g., about −20° C.).” (par [0072]).
Hanson further teaches a kit containing a reagent solution “disposed in a container.” Hanson, para. [0073].
Shikimura, Hanson, and Cathcart are analogous art because they relate to biological-assay systems in which liquid reagents are prepared, stored, transferred, and added to biological samples. It would have been obvious to one of ordinary skill in the art to load a premeasured quantity of Shikimura’s PMA-containing stimulant solution into the internal space of Cathcart’s pipette tip and store the loaded solution frozen, as taught by Hanson. One of ordinary skill in the art would have been motivated to preserve the stimulant as a stable, premade, ready-to-use reagent and permit the premeasured stimulant to be delivered directly from the pipette tip to Shikimura’s biological sample, as suggested by Hanson’s frozen ready-to-use reagent solution and Cathcart’s liquid-transfer pipette tip.
Regarding claim 8, Shikimura in view of Hanson and Cathcart teaches the measurement unit of claim 7 as discussed above.
Regarding “a temporary housing space is present between the second solution and the second opening,” Cathcart teaches:
“For liquids with low viscosity, such as water, it is frequently desirable to aspirate an air gap at the pipette tip after aspirating a volume of liquid, so movement of the pipette by the robot does not cause liquid to be dislodged from the pipette.” (col. 21, lines 28-32).
Because Cathcart aspirates the air gap through the pipette-tip opening after the liquid has been aspirated, the subsequently aspirated air moves the liquid inward and occupies a space between the liquid and the pipette-tip opening. Cathcart’s air gap therefore constitutes the claimed temporary housing space between the second solution and the second opening.
It would have been obvious to aspirate Cathcart’s air gap after loading Shikimura’s PMA-containing stimulant solution into the pipette tip and before freezing the solution according to Hanson. One of ordinary skill in the art would have been motivated to prevent the stimulant solution from being dislodged from or leaking through the pipette-tip opening during movement and handling, as expressly suggested by Cathcart.
Regarding claim 10, Shikimura in view of Hanson and Cathcart teaches the measurement unit of claim 7 as discussed above.
Regarding “the second container is a pipette tip,” Cathcart expressly teaches a liquid-handling apparatus having a “pipette tip” into which liquid is aspirated and from which the liquid is dispensed (col. 21, line 14-32).
Accordingly, Cathcart’s pipette tip corresponds to the claimed second container.
Regarding claim 14, Regarding “a second preparation step of preparing a second container that includes a second housing space and a second opening communicating with the second housing space,” Cathcart teaches preparing a pipette having a probe tip into which liquid is aspirated. Cathcart states:
“First, probe tip 33 is positioned over the surface 265 of a liquid to be aspirated, as shown in FIG. 10A. Next the tip is lowered to touch the surface, sensed by the capacitance sensing ability associated with the probe tip, as shown in FIG. 10B. Aspiration of a programmed amount is accomplished slowly, typically at about 1 micro-liter per second, while the tip is at the surface as shown in FIG. 10B.” (col. 20, line 61-68).
Cathcart further states:
“After the liquid is aspirated, the probe tip is slowly withdrawn…. The probe is then moved to wherever is required to dispense the liquid that has been aspirated.” (col. 21, lines 14-22).
Cathcart’s probe tip has an internal space that receives and retains the aspirated liquid and a distal opening through which the liquid is aspirated and dispensed. The internal space therefore corresponds to the second housing space, and the distal tip opening corresponds to the second opening communicating with the second housing space.
Regarding “a second solution containing a stimulant for activating a function of the biological sample,” Shikimura teaches:
“After 2.5 minutes, a protein kinase C activator (PMA (phorbol 12-myristate 13-acetate): neutrophil stimulator) was added to the biological sample to a concentration of 0.1 μM.” (par [0050]).
PMA is therefore a stimulant that activates a function of the biological sample, namely the myeloperoxidase activity and superoxide-production activity of neutrophils.
Shikimura and Cathcart do not expressly teach “a second freezing step of freezing [the] second solution . . . in a state of being housed in the second housing space.”
Hanson teaches providing an assay reagent as a premade solution and storing the solution frozen:
“It is preferred that the reagent solution be present in the form of a pre-made, ready to use reagent solution.” (par [0072]).
Hanson further teaches:
“The preferred reagent solution is stable at room temperature (e.g., about 25° C.), refrigerated (e.g., about 4° C.) or frozen (e.g., about −20° C.).” (par [0072]).
Hanson also teaches that the reagent solution is “disposed in a container.” (par [0073]).
Shikimura, Hanson, and Cathcart are analogous art because they relate to biological-assay systems in which reagent solutions are prepared, stored, transferred, and added to biological samples. It would have been obvious to one of ordinary skill in the art to aspirate a premeasured quantity of Shikimura’s PMA-containing stimulant solution into the internal space of Cathcart’s pipette tip and freeze the stimulant solution while it is retained in the pipette tip, in accordance with Hanson’s frozen-storage teaching. One of ordinary skill in the art would have been motivated to maintain the stimulant solution as a stable, premade, ready-to-use reagent, as suggested by Hanson, while allowing the premeasured stimulant to be delivered directly from Cathcart’s pipette tip to Shikimura’s biological sample.
Regarding claim 15, Regarding “in the second freezing step, the second solution is frozen in a state where a temporary housing space is present between the second solution and the second opening,” Cathcart teaches:
“For liquids with low viscosity, such as water, it is frequently desirable to aspirate an air gap at the pipette tip after aspirating a volume of liquid, so movement of the pipette by the robot does not cause liquid to be dislodged from the pipette.” (col. 21, lines 28-32).
Because the air is aspirated through the distal opening after the liquid has been aspirated, the subsequently aspirated air moves the liquid farther into the pipette tip and occupies a space between the liquid and the distal opening. Cathcart’s air gap therefore constitutes a temporary housing space present between the second solution and the second opening.
It would have been obvious to one of ordinary skill in the art to aspirate Cathcart’s air gap after aspirating Shikimura’s PMA-containing stimulant solution and then freeze the stimulant solution according to Hanson while the air gap remains between the solution and the opening. One of ordinary skill in the art would have been motivated to prevent the stimulant solution from being dislodged from the pipette tip during movement and handling, as expressly suggested by Cathcart.
Regarding claim 16, Shikimura in view of Hanson and Cathcart teaches the manufacturing method of claim 15 as discussed above.
Regarding “a solution introduction step of introducing the second solution into the second housing space before the second freezing step,” Cathcart teaches that:
“Aspiration of a programmed amount is accomplished slowly, typically at about 1 micro-liter per second, while the tip is at the surface.” (col. 20, line 65–68).
Cathcart further teaches that, after aspiration, the liquid is retained within the probe tip and transported to a location at which it is dispensed (col. 21, lines 13–22). Thus, Cathcart teaches introducing the liquid through the distal opening into the internal housing space of the pipette tip.
Regarding “in the solution introduction step, air is sucked into the second housing space via the second opening after the second solution is sucked into the second housing space via the second opening,” Cathcart expressly teaches:
“For liquids with low viscosity, such as water, it is frequently desirable to aspirate an air gap at the pipette tip after aspirating a volume of liquid.” (col. 21, lines 28-30).
Cathcart therefore teaches the claimed sequence: the liquid solution is first sucked through the pipette-tip opening into the tip’s internal housing space, and air is subsequently sucked through the same opening into that housing space. It would have been obvious to perform Cathcart’s aspiration sequence before freezing Shikimura’s stimulant solution according to Hanson because the sequence forms the temporary housing space recited in claim 15 and prevents the solution from being dislodged during subsequent handling.
Regarding claim 18, Shikimura teaches a biological sample measurement method for measuring a characteristic of a biological sample. (par [0021]).
Regarding:
“a preparation step of preparing the measurement unit according to claim 7,”
Shikimura teaches preparing an optical measurement container containing an indicator reagent solution and separately adding a PMA-containing stimulant solution.
Hanson teaches storing a premade indicator-containing assay solution frozen in a container, and Cathcart teaches housing and dispensing liquid through a pipette tip.
Thus, the references teach preparing the claim 7 measurement unit having the frozen first solution in the optical measurement container and the frozen second stimulant solution in the pipette-tip container, as discussed regarding claim 7.
Regarding:
“a first defrosting step of defrosting the first solution,”
Hanson teaches:
“It is preferred that the reagent solution be present in the form of a pre-made, ready to use reagent solution.”
Hanson further teaches:
“The preferred reagent solution is stable at room temperature (e.g., about 25° C.), refrigerated (e.g., about 4° C.) or frozen (e.g., about −20° C.).” (par [0072]).
Shikimura performs the assay using the reagent solution in liquid form at 37°C. Therefore, when Shikimura’s first reagent solution is stored frozen according to Hanson, the solution is defrosted before the biological sample is added and the optical assay is conducted.
Regarding:
“a first adding step of adding the biological sample to the first housing space via the first opening after the first defrosting step,”
Shikimura teaches:
“A biological sample was prepared by adding 0.5 μM of MCLA . . . 10 μM of APF . . . and CaCl₂ as 1 mM to RH buffer (10 mM HEPES, 154 mM NaCl, 5.6 mM KCl, pH 7.4) kept at 37° C., incubating the mixture at 37° C. for 4 minutes, and adding 3 μL of blood (whole-blood) collected from a target.” (par [0049]).
Thus, Shikimura adds the whole-blood biological sample through the opening of the sample container into the housing space containing the indicator reagent solution. In the modified method, this addition occurs after Hanson’s frozen first solution has been defrosted and brought to Shikimura’s liquid assay condition.
Regarding:
“a second defrosting step of defrosting the second solution,”
Shikimura teaches a second solution containing PMA as the neutrophil stimulant:
“After 2.5 minutes, a protein kinase C activator (PMA (phorbol 12-myristate 13-acetate): neutrophil stimulator) was added to the biological sample to a concentration of 0.1 μM.” (par [0050]).
As discussed regarding claim 7, Hanson’s frozen-storage teaching is applied to the PMA-containing solution housed in Cathcart’s pipette tip. The frozen PMA solution is defrosted before it is dispensed as a liquid from the pipette tip.
Regarding:
“a second adding step of leading out the second solution from the second opening and adding the second solution to the first housing space via the first opening after the second defrosting step,”
Shikimura teaches adding the PMA stimulant to the biological sample after measurement has begun, as quoted above.
Cathcart teaches that, after a liquid is aspirated into the pipette tip:
“The probe is then moved to wherever is required to dispense the liquid that has been aspirated.” (col. 21, lines 20-22).
Thus, Cathcart leads the liquid out through the opening of the pipette tip and dispenses it at a selected destination. Applying Cathcart’s dispensing operation to Shikimura’s defrosted PMA solution results in leading the stimulant solution from the second opening and adding it through the first opening into the first-container housing space containing the biological sample.
Regarding:
“a measurement step of measuring light generated in the first housing space and transmitted through the light transmission region of the first container,”
Shikimura teaches:
“The above-described irradiation of the biological sample was started while blinking an LED . . . as exciting light. After 2.5 minutes, a protein kinase C activator . . . was added to the biological sample . . . and the fluorescence intensity of APF (myeloperoxidase activity) and the chemiluminescence intensity of MCLA (superoxide-producing activity) were measured.” (par [0050]).
The APF fluorescence and MCLA chemiluminescence are generated within the sample-container housing space and pass through the light-transmitting container wall to the optical detector. Accordingly, Shikimura in view of Hanson and Cathcart teaches or suggests each step of claim 18.
Shikimura, Hanson, and Cathcart are analogous art because they relate to biological-assay methods involving indicator reagents, biological samples, optical measurements, and controlled delivery of liquid reagents. It would have been obvious to one of ordinary skill in the art to thaw the frozen, premade indicator and stimulant solutions taught by Hanson and then use Cathcart’s pipette dispensing operation to perform Shikimura’s disclosed sequence of adding blood, adding PMA, and measuring fluorescence and chemiluminescence. One of ordinary skill in the art would have been motivated to provide the assay reagents in stable, premeasured, ready-to-use forms, as suggested by Hanson, while accurately dispensing the stimulant into the biological sample using Cathcart’s liquid-handling technique.
Regarding claim 19, Regarding:
“a temperature adjustment step of adjusting a temperature of the first solution after the first adding step,”
Shikimura teaches that, after adding whole blood to the indicator-containing solution:
“The adjusted biological sample was incubated at 37° C. for 1 minute, and then measurement was started using a fluorescence and luminescence simultaneous measurement apparatus.” (par [0049]).
Thus, after the biological sample is added, Shikimura subjects the resulting first solution to a temperature-controlled incubation at 37°C.
Regarding:
“in the first defrosting step, the first solution is continuously heated such that the temperature of the first solution reaches an appropriate temperature for the biological sample,”
Hanson teaches storing the premade reagent solution frozen at approximately −20°C. (par [0072]).
Shikimura identifies 37°C as an appropriate assay temperature, teaching that the RH buffer is:
“kept at 37° C., incubating the mixture at 37° C. for 4 minutes.” (par [0049]).
It would have been obvious to continuously apply heat to Hanson’s frozen first solution until the solution is defrosted and reaches Shikimura’s disclosed 37°C assay temperature. Heating the solution without interruption until it reaches the selected assay temperature is a predictable method of converting the frozen reagent solution into the liquid, temperature-conditioned solution required by Shikimura.
Regarding:
“in the temperature adjustment step, the first solution is heated such that the temperature of the first solution is maintained at the appropriate temperature for the biological sample,”
Shikimura teaches maintaining both the reagent mixture and the biological sample at 37°C: the reagent mixture is incubated at 37°C before blood is added, and the adjusted biological sample is then incubated at 37°C after the blood is added (par [0049]).
Accordingly, one of ordinary skill in the art would have heated the frozen first solution until it reached 37°C and thereafter continued temperature control to maintain the first solution at 37°C after the biological sample was added. One of ordinary skill in the art would have been motivated to maintain the temperature selected by Shikimura so that the neutrophil assay is performed under Shikimura’s disclosed biological-assay conditions.
Claim 9 and 17 is rejected under 35 U.S.C. § 103 as being unpatentable over Shikimura in view of Hanson and Cathcart, and further in view of Solotareff.
Regarding claim 9, Shikimura in view of Hanson and Cathcart teaches the measurement unit of claim 8 as discussed above.
The combination does not expressly teach “a volume of the temporary housing space is equal to or larger than a volume of the second solution.”
Solotareff teaches a programmable pipette in which the aspirated-air volume and aspirated-liquid volume are user-selectable parameters. Solotareff states:
“Air aspiration. A sequence of air intake volumes through the pipette 30 are defined…. [T]he pipetting module 242 displays … a value of the volume of air to aspirate.” (par [0107])
Solotareff further teaches:
“Dilution. The user may specify the following parameters: the volume of aspirated air or the air gap between each liquid sample, [and] the volume of each liquid to aspirate….” (par [0110]).
Thus, Solotareff teaches that the air-gap volume and liquid volume are independently selectable variables. Cathcart expressly teaches that the air gap is provided to prevent the aspirated liquid from being dislodged from the pipette. It would have been obvious to one of ordinary skill in the art to select Solotareff’s adjustable air-gap volume relative to the volume of Shikimura’s stimulant solution to provide sufficient separation between the stimulant solution and the pipette-tip opening. Selecting an air-gap volume equal to or larger than the stimulant-solution volume would have been a routine optimization of known, independently adjustable, result-effective variables to more reliably retain the stimulant solution within the pipette tip during handling and frozen storage.
Regarding claim 17, Shikimura in view of Hanson and Cathcart teaches the method for manufacturing a measurement unit according to claim 16 for the reasons discussed above.
Regarding:
“a volume of the air sucked in the solution introduction step is equal to or larger than a volume of the second solution sucked in the solution introduction step,”
Solotareff teaches a programmable electronic pipette in which the volume of aspirated air is a selectable parameter:
“Air aspiration. A sequence of air intake volumes through the pipette 30 are defined. During this procedure, the pipetting module 242 displays on display 232 a name, a value of the volume of air to aspirate, and a speed of aspiration.” (par [0072]).
Solotareff further teaches that the air-gap volume and the liquid volume are separately selectable:
“Dilution. The user may specify the following parameters: the volume of aspirated air or the air gap between each liquid sample, the volume of each liquid to aspirate, the aspiration speed of each liquid, a name of each of the two to five liquids intended to be displayed in the display 170 of the pipette 30, and the name of the task.” (par [0110]).
Thus, Solotareff teaches independently specifying both the volume of the air gap and the volume of the aspirated solution.
Cathcart teaches the purpose of providing the air gap:
“For liquids with low viscosity, such as water, it is frequently desirable to aspirate an air gap at the pipette tip after aspirating a volume of liquid, so movement of the pipette by the robot does not cause liquid to be dislodged from the pipette.” (col. 21, lines 28-32).
Shikimura, Hanson, Cathcart, and Solotareff are analogous art because they relate to biological-assay reagent handling and the controlled aspiration, storage, and dispensing of liquid reagents. It would have been obvious to one of ordinary skill in the art to use Solotareff’s independently programmable air and liquid volumes when forming Cathcart’s air gap after aspirating Shikimura’s stimulant solution. Selecting the aspirated-air volume to be equal to or larger than the stimulant-solution volume would have been a routine adjustment of Solotareff’s disclosed programmable volumes to provide a sufficiently large separation between the stimulant solution and the pipette-tip opening. One of ordinary skill in the art would have been motivated to improve retention of the stimulant solution in the pipette tip during handling, as suggested by Cathcart’s express teaching that the air gap prevents the aspirated liquid from being dislodged.
Claim 20 is rejected under 35 U.S.C. § 103 as being unpatentable over Shikimura in view of Hanson and Cathcart, and further in view of Nakata.
Regarding claim 20, Shikimura in view of Hanson and Cathcart teaches the biological-sample measurement method of claim 19 for the reasons discussed above. The combination does not expressly teach:
“in the preparation step, a rotor immersed in the first solution is prepared, and in the temperature adjustment step, the first solution is heated while the rotor is rotated.”
Nakata teaches an optical biological-sample measurement apparatus having a rotor disposed directly in the reagent-containing sample solution:
“Reference numeral 7 denotes a rotor that allows the sample solution S contained in the sample container 5. The rotor 7 is housed in the sample container 5 along with the sample solution S and an emission reagent, and is rotationally driven by a magnetic stirrer 27 incorporated in the measurement section 6 when the sample container 5 is housed in the measurement section 6.”
Nakata further teaches:
“Thus, the number of microorganisms in the sample solution S can be counted while the sample solution S containing the sample and emission reagent in the sample container 5 is being stirred and mixed at a predetermined temperature.” (par [0063]).
Nakata’s rotor is housed within and immersed in the sample solution and emission reagent. Nakata also rotates the rotor while the solution is maintained at a predetermined temperature. When Nakata’s rotor is incorporated into the Shikimura-Hanson first container, the rotor is prepared immersed in the first solution. Performing Nakata’s rotation during Shikimura’s 37°C temperature-adjustment step results in heating the first solution while the rotor is rotated.
Shikimura, Hanson, Cathcart, and Nakata are analogous art because they relate to biological-sample measurement methods using reagent-containing sample containers and optical detection. It would have been obvious to one of ordinary skill in the art to place Nakata’s rotor in the first container of the frozen-reagent neutrophil-activity measurement method and rotate the rotor while the solution is heated and maintained at Shikimura’s 37°C assay temperature. Nakata expressly suggests the benefit of this modification, explaining that, compared with measuring a stationary solution, stirring and mixing:
“allows the microorganisms to emit light in a very short time, enabling the amount of the microorganisms . . . to be easily and quickly measured.” (par [0063]).
One of ordinary skill in the art therefore would have been motivated to improve mixing of the biological sample and indicator reagent during the temperature-controlled assay, thereby facilitating rapid and consistent optical measurement, as suggested by Nakata.
Response to Arguments
Applicant's arguments filed 09/17/2026 have been fully considered but they are not persuasive.
Applicant first notes that the Office Action refers to JP 2019-213464 as “Shikimura” and contends that the reference should instead be referred to as Kazumura. Examiner notes that the reference is properly identified as Shikimura et al. (JP 2019-213464 A).
Claims 1–3 and 11–12
Applicant argues on pages 8-10 that the combination of Shikimura and Hanson fails to teach or suggest the amended claim 1 limitation requiring:
“a first solution that is a mixture of a plurality of reagents containing an indicator reacting with a component generated from the biological sample, the indicator being an indicator reacting with hypochlorous acid or a halogen equivalent thereof produced by myeloperoxidase, or an indicator reacting with superoxide, wherein the first solution is housed in the first housing space in a state of being frozen.” (remark, page 7-8).
Applicant contends that Shikimura is directed to immediate measurement in a liquid state and does not disclose frozen storage. Applicant further argues that Hanson’s resazurin reagent is fundamentally different from the indicators recited in the claims because resazurin detects metabolic reduction, whereas the claimed indicators detect reactive oxygen species such as hypochlorous acid and superoxide. Applicant additionally argues that APF and MCLA have different stability characteristics from resazurin and that a person of ordinary skill in the art would not have had a reasonable expectation of success in applying Hanson’s frozen-storage teaching to Shikimura. (remark, page 8-10).
Examiner respectfully disagrees.
First, Applicant’s arguments regarding the difference between Hanson’s resazurin and the claimed indicators are not persuasive because Hanson is not relied upon for teaching the claimed indicator. Shikimura itself teaches the claimed indicator system. Specifically, Shikimura teaches use of APF and MCLA in a biological sample measurement method. Shikimura teaches that APF is an indicator that reacts with hypochlorous acid (or a halogen equivalent thereof) produced by myeloperoxidase (para. [0026] [0027]). Shikimura further teaches that MCLA is used to detect superoxide generated from the biological sample (para. [0028]). Accordingly, Shikimura teaches the claimed indicator reacting with hypochlorous acid produced by myeloperoxidase and/or the claimed indicator reacting with superoxide.
Therefore, the rejection does not rely on substituting Hanson’s resazurin indicator for Shikimura’s APF/MCLA indicator system. Rather, Shikimura provides the claimed indicator chemistry, and Hanson is relied upon only for the separate teaching of storing a premixed biological assay reagent solution in a frozen state.
Second, Applicant acknowledges that Hanson teaches a:
“pre-made, ready-to-use reagent solution” containing resazurin that can be stored frozen at about −20°C. 18641606-REM_2026-09-17
Hanson therefore demonstrates that freezing a premixed biological assay reagent solution is a known technique for storage and preservation of assay reagents. It would have been obvious to one of ordinary skill in the art to apply this known storage technique to Shikimura’s premixed APF/MCLA reagent solution in order to provide a prepared reagent solution that could be stored until needed and subsequently thawed before performing Shikimura’s biological measurement.
Applicant argues that APF and MCLA allegedly have different chemical stability and storage characteristics from Hanson’s resazurin. Applicant cites Exhibits indicating that MCLA and APF may have limitations when stored in aqueous solution. (remark, page 9).
However, Applicant has not established that freezing Shikimura’s reagent solution would have been unsuccessful or that one of ordinary skill in the art would have been discouraged from applying a known freezing-storage technique to Shikimura’s reagent solution. The cited evidence merely identifies certain properties of APF and MCLA; it does not establish that frozen storage of the claimed reagent mixture would have been technically infeasible.
Moreover, the proposed combination does not require maintaining Shikimura’s measurement solution in a frozen state during measurement. The proposed modification merely stores the premixed reagent solution in a frozen state before use. After thawing, the reagent solution would be used in the same manner as taught by Shikimura, including adding the biological sample and measuring the resulting fluorescence and/or chemiluminescence.
Applicant further argues that Shikimura itself provides no motivation to freeze the reagents because Shikimura performs measurement in a liquid state at 37°C. (remark, page 10).
Examiner respectfully disagrees. The fact that Shikimura performs the assay using a liquid reagent does not teach away from storing the reagent in a frozen state before use. A person of ordinary skill in the art would have understood that storage conditions for biological reagents need not be identical to assay operating conditions. Hanson provides an express teaching that premixed assay reagent solutions may be stored frozen and later used in a biological assay.
Accordingly, Applicant has not shown that the combination of Shikimura and Hanson would have lacked a reasonable expectation of success or that the rejection is based on improper hindsight. Applicant’s arguments are therefore not persuasive.
Claims 4 and 13
Applicant argues that Nakata fails to teach or suggest:
“a support that protrudes into the first housing space and rotatably supports the rotor.” (remark, page 11).
Applicant contends that Nakata merely discloses a magnetic rotor positioned within a sample container and rotated by an external magnetic stirrer, but does not disclose a support protruding into the housing space for rotatably supporting the rotor. Applicant further argues that none of the references recognize the problem of maintaining rotor positioning during freezing. (remark, page 11).
Examiner respectfully disagrees.
Nakata teaches the use of a rotor housed within the sample container and immersed in the reagent-containing solution. Specifically, Nakata teaches:
“The rotor 7 is housed in the sample container 5 along with the sample solution S and an emission reagent, and is rotationally driven by a magnetic stirrer 27 incorporated in the measurement section 6 when the sample container 5 is housed in the measurement section 6.” (par [0063]).
Thus, Nakata teaches the claimed arrangement of a rotor located within the solution for mixing the biological sample and reagent.
Although Nakata rotates the rotor through magnetic coupling with an external magnetic stirrer, it would have been obvious to provide a protruding support within the sample container to rotatably support the rotor. Providing a support for a rotating stirring element within a liquid container is a well-known mechanical arrangement. For example, magnetic stirring vessels commonly employ a bottom projection, bearing, or similar support structure to maintain the stirring bar at a desired location and reduce friction during rotation.
One of ordinary skill in the art would have been motivated to incorporate such a support into Nakata's sample container to improve stability of the rotor position and promote reliable rotation during mixing. The modification would not change Nakata's intended function because the rotor would continue to rotate within the solution to mix the sample and reagent. Rather, the modification merely provides a known mechanical support arrangement for implementing Nakata's disclosed rotor-based mixing operation.
Further, Applicant's argument concerning freezing does not establish non-obviousness. Hanson teaches storing a premixed reagent solution in a frozen state. Once the rotor and support are incorporated into the sample container before freezing, the rotor would remain immersed in the frozen reagent solution and available for rotation after thawing.
Conclusion
THIS ACTION IS MADE FINAL. 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 XIAOYUN R XU, Ph. D. whose telephone number is (571)270-5560. The examiner can normally be reached M-F 8am-5pm.
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/XIAOYUN R XU, Ph.D./ Primary Examiner, Art Unit 1797
