The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on October 23, 2025 has been entered.
The drawings are objected to under 37 CFR 1.83(a). The drawings must show every feature of the invention specified in the claims. Therefore, the sampling device configured for discharging sucked test blood sample or reagent to the reaction cell through suction and discharge actions must be shown or the feature(s) canceled from the claim(s). No new matter should be entered.
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. The figure or figure number of an amended drawing should not be labeled as “amended.” If a drawing figure is to be canceled, the appropriate figure must be removed from the replacement sheet, and where necessary, the remaining figures must be renumbered and appropriate changes made to the brief description of the several views of the drawings for consistency. Additional replacement sheets may be necessary to show the renumbering of the remaining figures. 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.
With respect to the claims, examiner attempted to determine what constitutes a “large platelet”.
Paragraphs [0053]-[0054] of the instant specification teach that for small platelets (such as platelets with a volume of less than 10 fL), due to small volumes, the platelets are further reduced in volume after being treated with the hemolytic agent. Furthermore, the inventors believe that after a blood sample is hemolyzed, the volumes of platelets are decreased, but the relative sizes are still kept unchanged, that is, the volumes of platelets that are originally large in volume after hemolysis are still large, the platelets that are large in volume can be detected in hemolytic channel through flow cytometry. Instant paragraph [00101] discusses figures 7A-7D teaching that they show data with respect to large platelets with volumes above 10 fL, 12 fL 15 L and 20 fL. There is no distinction relative to the volume and the name that might be used or why one might be interested in a platelet that is larger than any of the stated sizes. There is basis in this paragraph for the size information added to claim 38, but no specific reason for choosing any of the volume thresholds. Thus according to the instant disclosure a large platelet is simply one that is larger in size than the small platelets. For examination purposes, examiner will look for language indicative of a large platelet that has a volume above 20 fL.
Paragraph [0066] of the previously cited and below applied Nishimori reference (US 2014/0149938) provides examples of the blood cells to be displayed in an object scattergram. These include, similar to a conventional scattergram, red blood cells (including proerythroblasts which are a premature stage of red blood cells, basophillic erythroblasts, polychromatic erythroblasts, orthochromatic erythroblasts, reticulocytes, and abnormal forms thereof), platelets (including abnormal forms such as large platelets, giant platelets, and the like), and leukocytes (including neutrophils, eosinophils, basophils, lymphocytes, monocytes, atypical lymphocytes, abnormal lymphocytes, large lymphocytes, juvenile cells, and the like). From this it appears that large platelets and/or giant platelets are recognized terms in the art that were normally displayed on scattergrams.
The previously cited and below applied Oguni reference (US 2005/0002826) teaches apparatus and method for measuring immature platelets. Paragraph [0024] teaches the finding of a fluorescent dye which produces different staining characteristics between immature platelets and mature platelets, and which is excitable by a laser beam emitted from a semiconductor light source. An assay sample can be prepared using a staining reagent containing the fluorescent dye to detect optical information emitted from the assay sample that can be used to classify and count the immature platelets (this concept includes reticulated platelets). Paragraph [0055] describes figure 7 as an example of the two-dimensional scattergram prepared. This two-dimensional scattergram plots the forward scattered light intensity on the vertical axis, and the fluorescent light intensity on the horizontal axis. The region IP in which the immature platelets appear, and the region MP in which the mature platelets appear, are set. The immature platelets have a greater RNA content within the cell than do mature platelets, and they are larger than the mature platelets. Therefore, the region IP is set at a higher position of forward scattered light and fluorescent light intensities compared to the region MP. From this it is clear that immature platelets such as reticulated platelets are considered larger than mature platelets and could be considered as reticulated platelets for examination purposes. It is also clear that one of ordinary skill in the art is capable of producing a scattergram of fluorescent intensity and forward scattered light intensity and determining areas related to small and large platelets based on size (scatter) and fluorescent intensity (RNA content) for counting purposes.
The newly cited and below applied Merchez reference (US 2012/0296570) teaches a method of classifying and flow measuring the refringence of at least two populations of particles present in a fluid. Paragraph [0013] that the ability to distinguish between platelets of large size (macroplatelets) and microcytes (which are red blood cells of very small size), is important in order to avoid making certain errors of diagnosis. Unfortunately, among hematology analyzers other than those based on lasers, there is at present no solution that is simple and effective for avoiding this potential confusion between macroplatelets and microcytes. Paragraph [0089] teaches that the method includes a sub-classification step of classifying normal platelets, activated platelets, microplatelets, and macroplatelets. Since the concept of a "macroplatelet" relates to volume, the sub-classification may be adapted for each laboratory, as with erythrocytes. Paragraph [0090] teaches that the step of classifying events separates populations of leukocytes as constituted in particular by lymphocytes, monocytes, granulocytes, neutrophils, and eosinophilic granulocytes. Paragraphs [0299]-[0300] teach that the platelets are sub-classified in order to distinguish between activated platelets, small platelets, and macroplatelets. This produces four sub-populations of platelets as can be seen in figure 20. If their volume is less than 20 fL, then they are small platelets, and if it is greater than 20 fL, they are macroplatelets. Figures 22A and 22B compare classifications for a case of macroplatelets showing the region of the scattergram that is selected. Thus Merchez shows that one of ordinary skill in the art can identify regions of a scattergram that reflect the presence of large platelets (macroplatelets) having a volume greater than 20 fL and that knowledge of the number of macroplatelets is an important property to determine/measure.
With respect to the newly added sampling device language in claim 38, examiner notes that its principle description is found in instant paragraph of the originally filed specification. Of particular interest is the fact that it is not shown in the drawings. Also of note is the fact that the description covers just about any structure capable of adding sample and the other agents mixed therewith to the reaction cell. Thus as long as there is a reaction cell for mixing sample with a lysing agent and a fluorescent dye, the sampling device language of claim 38 will be considered as met.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 38-41, 46-51 and 56-60 are rejected under 35 U.S.C. 103 as being unpatentable over Bo (WO 2016/106688 or US 2018/0003634, both newly cited, applied and publications based on PCT/CN2014/095905 so that a description of the English language publication US 2018/0003634 will be appropriate for both publications) in view of Narisada (US 2003/0032193), Sakata (US 5,496,734), Merchez (US 2012/0296570, newly cited and applied) and Oguni (US 2005/0002826) or Nishimori (US 2014/0149938). The Bo publications teach a device and method for warning about the presence of nucleated red blood cells using flow cytometers. The warning device may obtain forward-scattered light information, side-scattered light information and fluorescence information when cells in the blood sample pass through a detection region of the flow cytometer. The warning device may generate a side-scattered light-fluorescence dot plot, so as to classify leucocytes into four groups, and may generate a forward-scattered light-fluorescence dot plot, where the forward-scattered light-fluorescence dot plot can include a leucocyte group region. The warning device may obtain a predetermined feature region located at the left side of the leucocyte group region, perform statistics on the amount of characterization cells in the predetermined feature region, and provide a warning when the number of the characterization cells exceeds a threshold value. Figures 2A, 3A, 5A and 6A show the generated side-scattered light-fluorescence scattergram (dot plot) used to classify leucocytes into four groups (a lymphocyte population, a monocyte population, an eosinophil population, and a neutrophil and basophil population). Figures 2B, 3B, 5B, 6A and 12 show the generated a forward-scattered light-fluorescence scattergram (dot plot). Paragraph [0056] describing figure 1 teaches that at step S10, a flow cytometer and a blood sample containing leucocytes may be provided, where the blood sample may be preprocessed to obtain a processed blood sample. In some examples, the blood sample may be mixed in proportion with a reagent containing a fluorescent dye and a hemolytic component to form the processed blood sample. The reagent may be used to hemolyze red blood cells in the blood sample, so as to avoid interference to the counting of the leucocytes and the nucleated red blood cells. Also, the fluorescent dye in the reagent may bind with nucleic acids in the leucocytes and the nucleated red blood cells to label those cells, where since different types of cells may have different binding capabilities to the fluorescent dye, different fluorescence information may be produced. Moreover, since various types of cells may have different sizes, different forward-scattered light information may be produced, and different side-scattered light information may be produced due to different intracellular morphologies or complexities. It may be understood that the flow cytometer may be used to cause the cells to pass through a detection region one by one to collect the information above, and then various types of cells can be distinguished on a dot plot through analysis. Paragraph [0107] describes figure 11 as showing a schematic structural diagram of the detection device 20. The detection device can include: a light source 1025, a flow chamber 1022 serving as the detection region, a forward-scattered light collection device 1023 provided on an optical axis, and a side-scattered light collection device 1026 and a side-scattered fluorescence collection device 1027 provided by a side of the optical axis. The cells of the blood sample processed by the preprocessing device (30, figure 10) may successively enter the flow chamber (the detection region) via a tube 3, where the forward-scattered light collection device, the side-scattered light collection device and the side-scattered fluorescence collection device in the same channel may successively detect and collect the forward-scattered light, the side-scattered light and the side-scattered fluorescence information of each cell, and may transmit these information to the analysis device (40, figure 10) via a communication interface 4. Paragraph [0103] teaches that the preprocessing device 30 may preprocess the blood sample so as to obtain a processed blood sample, where the preprocessing may at least include performing fluorescence labeling on cells in the blood sample. Paragraphs [0105]-[0106] teach that the analysis device 40 can be implemented in a CPU. The analysis device 40 may generate a forward-scattered light-fluorescence dot plot of the blood sample according to the forward-scattered light information, the side-scattered light information and the fluorescence information detected by the detection device, acquire a predetermined feature region of the forward-scattered light-fluorescence dot plot, perform statistics on the amount of characterization cells in the predetermined feature region, determine whether the amount of the characterization cells exceeds a threshold value, and provide a warning when it is determined that the amount of the characterization cells exceeds the threshold value. Here, the forward-scattered light-fluorescence dot plot may include a leucocyte population region, and the predetermined feature region may be located at the left side of the leucocyte population region, where the location and the size of the predetermined feature region may be fixed or dynamically set. Paragraphs [0091]-[0092] gives additional description on how the processor (CPU) is configured to produce the different dot plots and statistically analyze the predetermined feature region. Paragraph [0061] teaches that the location and the size of the predetermined feature region may be unchanged (i.e. decided in advance), or may be dynamically adjusted as a function of the position of the leucocyte population or a blood shadow region in the forward-scattered light-fluorescence dot plot. In some examples, two forward-scattered light-fluorescence dot plots may be obtained by respectively detecting a normal blood sample containing no nucleated red blood cells and an abnormal blood sample known to contain nucleated red blood cells in the same detection system by means of the above-mentioned method. Through comparison, a region may be found at the left side of the leucocyte population, in which case a particle population appears in this region of the forward-scattered light-fluorescence dot plot of the abnormal blood sample, while no particle population exists in this region of the forward-scattered light-fluorescence dot plot of the normal blood sample. In this way, this region can be determined as the predetermined feature region. Paragraphs [0068]-[0069] teach ways that the predetermined feature region can be dynamically adjusted. Paragraph [0030] teaches that in order to detect nucleated red blood cells, a specialized detection reagent and method may be used aiming at the nucleated red blood cell. Although this method using a specialized reagent system and detection device can count the nucleated red blood cells, it may lead to increased volume and complexity of detection instruments, and the costs of clinical examination may also be increased. Paragraph [0110] teaches that in the embodiments of this disclosure, a warning for nucleated red blood cells can be provided while classifying and detecting leucocytes without using specialized nucleated red blood cell reagent and without counting the amount of the nucleated red blood cell, whereby information for clinical screening can be provided quickly, simply and conveniently without increasing complexity and costs of the measurement. Paragraphs [0112]-[0113] teach that the technical features or operation steps illustrated in the embodiments of this disclosure can be combined in any way. Those of ordinary skill in the art may easily understand that the sequence of steps or actions in the methods illustrated by the embodiments of this disclosure can be altered. Therefore, unless a certain sequence is specified otherwise, any sequence in the figures or the detailed description is merely for the purpose of illustration and not an obligatory sequence. The descriptions provided are merely preferred implementations of this disclosure and should not be taken as limiting of the disclosure. Based on the above description, relative to claim 38, Bo teaches a blood analyzer, comprising: a test sample preparation apparatus for preparing a test sample for measurement, wherein the test sample preparation apparatus at least comprises a sampling device and a reaction cell (the sample is mixed with a reagent and provided to the flow chamber of figure 11 through flow line 3), wherein the sampling device is configured for discharging sucked test blood sample or reagent to the reaction cell through suction and discharge actions and the reaction cell is configured for providing a place for mixing and incubation of the test blood sample and the reagent (see two embodiments of the process described in paragraphs [0074]-[0082]); a flow chamber communicating with the reaction cell through a pipeline (see elements 3 and 1022 of figure 11); a measurement apparatus comprising a light source and an optical detection device, wherein the light source is configured for emitting a light beam to irradiate the flow chamber, and the optical detection device is configured for receiving scattered light and fluorescent light See the description of figure 11 in paragraph [0107]); and a data processor configured for: causing the sampling device to collect a test blood sample and injecting a certain amount of the test blood sample into the reaction cell; causing the sampling device to add a fluorescence staining agent and a hemolytic agent to the reaction cell, so that the test blood sample, the fluorescence staining agent and the hemolytic agent are mixed and incubated in the reaction cell, which causes red blood cells in the test blood sample to be lysed and causes cell particles in the sample to be stained, so as to prepare the test sample (see at least paragraphs [0056] and [0074]-[0082]); flowing, through fluid path control, a certain amount of the test sample in the reaction cell to the flow chamber, so that cell particles in the test sample are passed through the flow chamber one by one under the irradiation of the light source (see at least figures 2A-2B, 3A-3B, 5A-5B and 6A-6B along with paragraphs [0056] and [0074]-[0082]); causing the optical detection device to receive first angle scattered light, second angle scattered light and fluorescent light generated by the cell particles passing through the flow chamber under the irradiation of light and to output first angle scattered light information, second angle scattered light information and fluorescent light information, wherein the first angle scattered light information is used for reflecting volume size information of the cell particles and the second angle scattered light information is used for reflecting complexity information of the cell particles (see at least figures 2A-2B, 3A-3B, 5A-5B and 6A-6B along with paragraphs [0027]; a scattered light signal and a fluorescence signal thereof reflect physicochemical features of the cells, such as the size, granularity and the expression of antigen molecules of the cells, [0056] and [0074]-[0082]); and classifying white blood cells in the test sample as lymphocytes, monocytes, neutrophils and eosinophils, according to the second angle scattered light information and the fluorescent light information of the cell particles (see at least figures 2A, 3A, 5A and 6A along with paragraphs [0056] and [0074]-[0082]). Bo also teaches identifying and/or classifying other particles of interest using a predetermined region in a dot plot of first angle scattered light information and fluorescent light information of the cell particles although those particles are not reticulocyte particles or large platelets, wherein the data processor is further configured for generating a scatter diagram at least according to the first angle scattered light information and the fluorescent light information, obtaining a first region and a second region from a blood ghost region of the scatter diagram, and identifying particles in the first region as the reticulocyte particles, and identifying particles in the second region as large platelets larger than 20fL.
With respect to claim 38, Narisada teaches a particle analyzer which includes a detecting section for detecting respective characteristic parameters of a plurality of particles, a distribution map preparing section for preparing at least two kinds of two-dimensional frequency distribution maps of the particles by using the detected characteristic parameters, a classifying section for classifying the particles into particle clusters on the two kinds of distribution maps, a calculating section for calculating and comparing the respective numbers of particles in the particle clusters containing particles of common kind to the two kinds of distribution maps, and a judging section for judging a classification error on the distribution maps based on a comparison result. Specifically with respect to claim 38, Narisada teaches a test sample preparation apparatus for preparing a test sample for measurement (see figure 2), wherein the test sample preparation apparatus at least comprises a reaction cell (48,51,52,53), and the reaction cell is configured for providing a place for mixing and incubation of a test blood sample, a fluorescence staining agent and a hemolytic agent, and the mixing and incubation causes red blood cells in the test blood sample to be lysed and causes cell particles in the sample to be stained (see at least paragraph [0038], reaction chamber 48 in which the sample liquid is reacted with a reagent); a flow chamber configured for providing an area for the cell particles in the test sample to pass through one by one and to be irradiated with light (sheath flow cell 1, see at least figure 1); a measurement apparatus comprising a light source (21) and an optical detection device, wherein the light source is configured for emitting a light beam to irradiate the flow chamber, the optical detection device is configured for receiving scattered light and fluorescent light generated by the cell particles under the irradiation of light and outputting scattered light information and fluorescent light information (photomultiplier tube 31, see at least figure 1 with its associated discussion), and the scattered light information at least comprises first angle scattered light information which is used for reflecting volume size information of the cell particles (photodiode 26, see at least figure 1 with its associated discussion) and second angle scattered light signal which is used for reflecting complexity information of the cell particles (photomultiplier tube 29, see at least figure 1 with its associated discussion); and a data processor (analysis section 35, also see figure 3 with its associated discussion) configured for identifying reticulocyte particles according to the first angle scattered light information and the fluorescent light information of the cell particles (figure 7 shows a two-dimensional frequency distribution map with a cluster of reticulocytes, a cluster of matured erythrocytes and a cluster of platelets that are respectively classified) and classifying white blood cells in the test sample as lymphocytes, monocytes, neutrophils and eosinophils according to the second angle scattered light information and the fluorescent light information of the cell particles (figure 6 shows a two-dimensional frequency distribution map having a cluster of lymphocytes, a cluster of monocytes, a cluster of neutrophils and basophils and a cluster of eosinophils that are respectively classified). With respect to claim 39, the data processor is configured for generating a scatter diagram at least according to the first angle scattered light information and the fluorescent light information of the cell particles, and identifying particles in a first region of the scatter diagram as reticulocyte particles, wherein the first region refers to a region in a blood ghost region (see figures 7 and 9 and paragraphs [0060]-[0062]). With respect to claim 40, the data processor is further configured for acquiring reticulocyte information according to the identified reticulocyte particles (see at least figure 9 step S8d and paragraphs [0060]-[0062] and [0069]). With respect to claim 41, the reticulocyte information comprises at least one of the followings: reticulocyte marker information, reticulocyte count information, high fluorescent reticulocyte count information, middle fluorescent reticulocyte count information, low fluorescent reticulocyte count information, immature reticulocyte count information, and nucleic acid content in reticulocytes (see at least figure 9 step S8d and paragraphs [0060]-[0062] and [0069]). With respect to claims 44-47, see figures 4-7. Claims 48-51 and 54-57 are methods equivalent to use of the blood analyzer described above. Narisada describes a method of preparing and measuring reticulocytes in paragraphs [0060]-[0062]. Claim 58 is of a scope which is broader than claim 48 so that any combination of teachings showing the obviousness of claim 48 also covers claim 58 as well. With respect to claim 59, paragraphs [0057]-[0059] teach that a hemolytic agent in combination with a fluorescent stain are added to blood to hemolyze the erythrocytes and stain the leukocytes to prepare the two-dimensional frequency distribution map of figure 6 showing the required leukocyte groups so that the hemolytic agent does not affect the white blood cell measurement.
In the patent Sakata teaches a method for blood analysis which provides a rapid treatment of a blood sample so that it can be analyzed for counting and classification of leukocytes. Leukocytes are permeabilized by treatment with a surfactant solution and labeled, preferably with a fluorescent dye. The method provides labeled leukocytes that can be counted and classified by optical means, including flow cytometry by analysis of a fluorescence signal. The paragraph bridging column 2-3 teaches that the blood sample is treated with an aqueous solution comprising at least one surfactant selected from the group consisting of a cationic surfactant and an amphoteric surfactant, and with a labeling substance in which; the aqueous solution of the surfactant is used at a concentration that does not destroy the whole cell membrane of a leukocyte but is sufficient to slightly damage a part of the cell membrane so as to make it permeable. The labeling substance is any substance which is capable of passing through the damaged cell membrane and combining with a component contained in the treated leukocyte; and the treatment is performed at a pH of 3.0 to 11.0. Column 7 gives further details on the types of cationic and amphoteric surfactants which can be used and the concentration needed to damage the cell membrane so as to make it permeable. In particular, column 7, lines 56-67 teach that the presence of the surfactant above a certain concentration lyses cells. Thus the surfactant is considered a lysing or hemolytic agent. Column 9 teaches that a nonionic surfactant may be added to the aqueous reagent solution of the cationic or amphoteric surfactant and the labeling substance. The column also provides further details relative to the types and concentration of the nonionic surfactants that can be used. The paragraph bridging columns 9-10 discusses the function of the nonionic surfactant as controlling the action of the ionic surfactant toward the cell membrane, thereby inhibiting the lysis of cell components with the ionic surfactants. For example, the use of the ionic surfactant associated with the nonionic surfactant is preferable when the ionic surfactant has so potent a lysing activity that leukocytes are unnecessarily damaged. The nonionic surfactant also will act to accelerate the lysis of erythrocytes, which causes a problem in the case of measuring leukocytes contained in a blood sample. Further, it can act to solubilize substances which are precipitated from a reaction solution by neutralization of anionic substances included in cell components or others of the blood sample, with the cationic surfactant. In addition, it can exert to inhibit aggregation of erythrocytes which remain as ghosts without their complete lysing, which will occur in the case where erythrocytes are rich as in a test sample of blood product and the dilution of a said test sample with the aqueous reagent solution is obliged to be low. A further description of the function of the ingredients of the reagent composition is further described in column 10, line 63 to column 11, line 22 as follows. The surfactant is believed to function to remove a part of substances which constitute a cell membrane, probably lipid molecules, thereby yielding pores in cell membrane which can pass a substance which does not usually pass the membrane. This is the damage of the cell membrane through which a labeling substance such as a dye is allowed to come into a cell and rapidly combine with an intracellular component. As a secondary effect, the cationic and amphoteric surfactants having a positive charge in their molecule have a function that the positive charge is ionically bound with the intracellular components having negative charge (for example, RNA with a phosphoric group, protein with a carboxyl group, and the like), whereby the intracellular components become insoluble. The insolubilized components do not leak out from the cell even when its membrane is damaged. Moreover, the cell which accumulates the insolubilized components is prevented from leaking of most of its cytoplasm, nucleus, and granules. Several examples follow of which examples 3 and 4 are particularly relevant. In example 3, column 13, lines 1-13 teach a reagent composition which is added to blood and mixed. Following this, red fluorescent light, forward scattered light, and side scattered light were measured by a flow cytometer. Figures 17 and 20 along with their associated discussion as part of example 3, teach that for certain compositions of the blood sample treating solution a scattergram based on fluorescence and side scattered light is capable of classifying white blood cells into four groups that appear to be equivalent to those taught by Narisada. In example 4, a reagent composition including a cationic surfactant, a nonionic surfactant and a labeling substance is given in column 13, lines 40-55. Column 13, lines 56-62 teaches that this reagent composition was mixed with venous blood. After 20 seconds at a room temperature, the red fluorescence light, green fluorescence light, forward scattered light, and side scattered light were measured by a flow cytometer. The paragraph bridging columns 13-14 teach that the scattergram in figure 28 is a scattergram that shows measured forward scattered light (FSC) and red fluorescence light (RFL). The “De” label indicates reduced erythrocyte membrane and blood platelet (erythrocyte ghost). Figure 37 is a scattergram showing the relationship between the intensity of the forward scattered light (FSC) and the intensity of the red fluorescence light (RFL) when subjects with erythroblast and immature granulocytes is treated with the reagent composition of Example 4. Column 15, lines 7-20 teach that figures 35 to 37 show the results obtained from the samples in which erythroblasts (Er) and immature granulocytes (Im) were erupted. As seen from these figures, Erythroblasts (Er) and immature granulocytes (Im) were classified and counted. Since Blast cells (Er) and (My), Atypical lymphocytes (Aly), Immature granulocytes (Im) include RNA richly therein, they can be detected by staining RNA. In addition, Reticulocytes (Ret) were partially detected in figure 37. Examiner notes that figure 37 includes an unlabeled region similar to that labeled with “De” characterized as reduced erythrocyte membrane and blood platelet (erythrocyte ghost) in figure 28. Examiner also notes that it is similar to figures 2B, 3B, 5B, 6B and 12 of the primary Bo references. In this example, erythrocytes are not completely lysed, but rather, only the membrane of the erythrocyte was damaged. Therefore, cell membranes of the cells which are converted into ghosts and RNA of the cell components are stained, thereby detecting reticulocytes.
In the patent publication Merchez teaches devices and methods for performing flow measurements to characterize microparticles, and in particular biological cells. Paragraph [0013] that the ability to distinguish between platelets of large size (macroplatelets) and microcytes (which are red blood cells of very small size), is important in order to avoid making certain errors of diagnosis. Unfortunately, among hematology analyzers other than those based on lasers, there is at present no solution that is simple and effective for avoiding this potential confusion between macroplatelets and microcytes. Paragraph [0089] teaches that the method includes a sub-classification step of classifying normal platelets, activated platelets, microplatelets, and macroplatelets. Since the concept of a "macroplatelet" relates to volume, the sub-classification may be adapted for each laboratory, as with erythrocytes. Paragraph [0090] teaches that the step of classifying events separates populations of leukocytes as constituted in particular by lymphocytes, monocytes, granulocytes, neutrophils, and eosinophilic granulocytes. Paragraphs [0299]-[0300] teach that the platelets are sub-classified in order to distinguish between activated platelets, small platelets, and macroplatelets. This produces four sub-populations of platelets as can be seen in figure 20. If their volume is less than 20 fL, then they are small platelets, and if it is greater than 20 fL, they are macroplatelets. Figures 22A and 22B compare classifications for a case of macroplatelets showing the region of the scattergram that is selected. Thus Merchez shows that one of ordinary skill in the art can identify regions of a scattergram that reflect the presence of large platelets (macroplatelets) having a volume greater than 20 fL and that knowledge of the number of macroplatelets is an important property to determine/measure.
In the patent publication, Oguni teaches apparatus and method for measuring immature platelets. The apparatus includes (a) a sample preparation unit for preparing an assay sample by adding a reagent to a blood specimen; (b) a detection unit having a semiconductor laser light source for irradiating the assay sample with laser light, and a detector for detecting optical information emitted from particles within the assay sample irradiated by laser light; and (c) a controller for differentiating and counting immature platelets based on the detected optical information. Paragraph [0003] teaches that the measurement of reticulated platelets, that is immature platelets, is considered to reflect the platelet production function in the marrow, and has been reported to be useful in differentiating conditions such as idiopathic thrombocytopenic purpura (ITP) and other thrombopenic diseases (for example, aplastic anemia (AA)). Paragraph [0024] teaches a fluorescent dye was found which produces different staining characteristics between immature platelets and mature platelets, and which is excitable by a laser beam emitted from a semiconductor light source. An assay sample can be prepared using a staining reagent containing the fluorescent dye to detect optical information emitted from the assay sample that can be used to classify and count the immature platelets (this concept includes reticulated platelets). Paragraph [0029] teaches that the sample preparation unit (2) is provided with a dilution fluid container (21) for accommodating a dilution fluid used for diluting a specimen, a staining fluid container (22) for accommodating a staining fluid used for staining a specimen, and a reaction vessel (23) for mixing the dilution fluid, staining fluid, and a specimen. Paragraph [0033] teaches that a control unit (4) is provided with a microcomputer (40) which includes a central processing unit (400) and a memory (401), a control circuit (41) for controlling the operation of each unit of the apparatus for measuring immature platelets, and a signal processing circuit (42) for subjecting the forward scattered light signals and fluorescent light signals sent from the detection unit to a noise elimination process and extracting the required data. Stored in the memory are control programs for controlling the operation of each part of the apparatus for measuring immature platelets through the control circuit and executing the series of assay operations, and analysis programs for analyzing the extracted data processed by the signal processing circuit and counting the immature platelets and mature platelets contained in the specimen. The analysis results obtained by the analysis program are output to a liquid crystal touch panel (10). Paragraph [0036] teaches that the sample preparation unit is controlled to prepare an assay sample from the specimen and predetermined reagents. An amount of suctioned specimen, 4.5 µL, is discharged into the reaction vessel. Then, 0.8955 mL of dilution fluid is supplied from the dilution fluid container to the reaction vessel. Next, 18 µL of staining fluid is supplied from the staining fluid container to the reaction vessel. Thereafter, the fluids are mixed for 31 seconds to stain the diluted specimen. Paragraph [0037] teaches that the fluorescent dye in the staining fluid is capable of bonding with RNA in cells. Therefore, a difference in stainability occurs between cells having a large RNA content (for example, immature platelets) and cells having a low RNA content (for example, mature platelets), such that there is also a difference in the intensities of the fluorescence detected. Paragraph [0055] describes figure 7 as an example of the two-dimensional scattergram prepared. This two-dimensional scattergram plots the forward scattered light intensity on the vertical axis, and the fluorescent light intensity on the horizontal axis. The region IP in which the immature platelets appear, and the region MP in which the mature platelets appear, are set. The immature platelets have a greater RNA content within the cell than do mature platelets, and they are larger than the mature platelets. Therefore, the region IP is set at a higher position of forward scattered light and fluorescent light intensities compared to the region MP. Thus immature platelets are large platelets (larger than mature platelets or small platelets). Paragraphs [0069]-[0070] describe figure 14 as a two-dimensional scattergram obtained by assaying a blood specimen using the apparatus for measuring immature platelets. In addition to the region IP in which reticulated platelets appear, and the region MP in which mature platelets appear, a region RBC in which red blood cells appear and a region RET in which reticulocytes appear are also shown. In the scattergram, the region RBC and region RET are set at positions having greater forward scattered light intensity than the region IP and region MP. This condition is set because the red blood cells and reticulocytes are larger than the immature platelets and mature platelets. The region RET is set at a position of greater fluorescent light intensity than the region RBC. This condition is set because the reticulocytes, which are immature red blood cells, have RNA within the cells, and are more readily fluorescently stained than the mature red blood cells, which do not have RNA within the cells. In this embodiment, not only are the reticulated platelets and mature platelets classified and counted, the red blood cells and reticulocytes are also classified and counted.
In the patent publication Nishimori teaches a display device composed of at least a data processor and a display for displaying a scattergram on the display. Paragraph [0001] teaches that the display device is for displaying a scattergram (also called scatter diagram, scatter plot, scatter plot graph, and the like) showing the distribution state of cells, particularly blood cells. Paragraphs [0002]-[0009] describe the cell types (red blood cells, platelets, and leukocytes) depicted in the scattergrams and conditions related to the relative amounts of certain types. Also described is at least one kind of scattergram relating to four kinds of leukocytes (namely, Lymphocytes (L), Monocytes (M), Neutrophils (N), and Eosinophils (E)), stained using a cell staining technique such as fat staining, peroxidase staining, and the like. Paragraph [0066] that examples of the blood cells to be displayed as the object scattergram include, similar to conventional scattergram, red blood cells (including proerythroblasts which are a premature stage of red blood cells, basophillic erythroblasts, polychromatic erythroblasts, orthochromatic erythroblasts, reticulocytes, and abnormal forms thereof), platelets (including abnormal forms such as large platelets, giant platelets, and the like), and leukocytes (including neutrophils, eosinophils, basophils, lymphocytes, monocytes, atypical lymphocytes, abnormal lymphocytes, large lymphocytes, juvenile cells, and the like).
Relative to claims 38, 48 and 58, it would have been obvious to one of ordinary skill in the art to modify the device and method of Bo by adding predetermined feature regions in the areas identified by Sakata for reticulocytes and platelets and combining the detection of reticulocytes and large platelets or immature platelets as taught by Merchez, Oguni or Nishimori with the leucocytes using scattered light and fluorescence as taught by Sakata because of the ability and desire to detect and/or measure reticulocytes and platelets in/with the apparatus of Narisada having a structure and detection principle similar to Bo, the quickness, simplicity and convenience without increasing the complexity and cost of the measurement as taught by Bo and the ability to combine the detection of reticulocytes with immature or large platelets and leucocytes as taught by Merchez, Oguni, Nishimori and/or Sakata.
Claims 41-43 and 51-53 are rejected under 35 U.S.C. 103 as being unpatentable over Bo in view of Narisada, Sakata Merchez and Oguni or Nishimori as applied to claims 40-41 and 50-51 above, and further in view of Mizukami (US 2004/0132196). Bo teaches an analyzer that provides a warning when the particle detected in the predetermined feature region reaches a threshold value. Bo does not teach a similar alarm based on reticulocyte information or the measurement of high fluorescent reticulocyte count information, the middle fluorescent reticulocyte count information, and the low fluorescent reticulocyte count information according to fluorescent light signal intensity distribution of the reticulocyte particles.
In the patent publication Mizukami teaches methods and devices are for monitoring a therapeutic effect in a red blood cell-related disease by detecting the size of a reticulocyte using a full-automatic blood cell counting device that figure 1 shows is similar to the device of Bo in figure 11. Paragraphs [0003]-[0004] teach several diseases that are associated with increases or decreases in the reticulocyte count. Paragraph [0026] teaches that the size of a reticulocyte (and red blood cell) is proposed to relate closely to the level of hemoglobin contained in its cell (see, for example: "New Red Cell Parameters on the Sysmex XE-2100 as Potential Markers of Functional Iron Deficiency" by C. Briggs, R. Rogers, B. Thompson, S. J. Machin in Infus. Ther. Transfus Med. 2001, 28, 256-262). Thus, to investigate the size of a reticulocyte is to investigate the hemoglobin level in the reticulocyte in an indirect manner, whereby allowing a hypochromic red blood cell whose hemoglobin level is reduced to be detected. Paragraph [0033] teaches that the principles of measurement of a reticulocyte by flow cytometry are described below. First, RNA present in the reticulocyte is bound to a fluorescent dye to effect staining, and then the stained reticulocyte is allowed to flow into a flow cell of a flow cytometer. The stained reticulocyte is irradiated with an excitation light. A scattering light and a fluorescent light thus emitted are then detected by a detector. The wavelength of the excitation light may be selected appropriately depending on the fluorescent dye employed. Paragraph [0034] teaches that a two dimensional distribution diagram (scattergram) whose two axes correspond to lateral fluorescent light intensity and forward scattering light intensity, respectively, is produced from a forward scattering signal and a lateral fluorescent light signal among the signals detected by the detector. A mature red blood cell region and a reticulocyte region are specified by known methods. Since a mature reticulocyte contains no RNA in its cell, the resultant fluorescence is extremely low. A reticulocyte, which contains RNA, exhibits a relatively intense fluorescence. Thus, a cell having fluorescence at a certain level or higher may be regarded as a reticulocyte. Moreover, since fluorescent light intensity varies depending on the amount of RNA contained in a cell, the maturity of the reticulocyte can be determined on the basis of the resultant fluorescent light intensity. A higher fluorescent light intensity reflects a cell containing a higher amount of RNA, which corresponds to a more juvenile reticulocyte. In the case of a multiparameter automated hematology analyzer, such as the Model XE-2100 produced by SYSMEX CORPORATION, a reticulocyte is divided depending on its fluorescent light intensity into any of the three regions shown in figure 3: HFR (high fluorescence ratio), MFR (middle fluorescence ratio) and LFR (low fluorescence ratio). Most juvenile reticulocytes are located in the HFR region. Paragraph [0035] teaches that in contrast to conventional methods, upon measuring a reticulocyte, the method focuses on the size of a cell in addition to the fluorescent light intensity (i.e., RNA quantity). Since a forward scattering light intensity obtained by measuring a reticulocyte by flow cytometry reflects the size of the cell, it may be employed as information for the cell size. For example, a mature red blood cell region and a reticulocyte region are specified on the two dimensional distribution diagram (scattergram). The ordinate of the diagram corresponds to forward scattering light intensity and the abscissa of the diagram corresponds to lateral fluorescent light intensity. The specified reticulocyte region is subjected to determination of the mean forward scattering light intensity. After a patient is treated with a therapeutic agent, the reticulocyte in the blood of the patient is measured, and the change in the measured value is monitored at after certain time interval, thereby monitoring the therapeutic effect. In addition, the regions classified based on the maturity of the reticulocyte (HFR, MFR, LFR described above) are also subjected to a determination of mean forward scattering light intensities in the respective regions, thereby obtaining additional detail regarding the therapeutic effect.
It would have been obvious to one of ordinary skill in the art at the time the application was filed to incorporate the determination of high fluorescent reticulocyte count information, the middle fluorescent reticulocyte count information, and the low fluorescent reticulocyte count information according to fluorescent light signal intensity distribution of the reticulocyte particles as taught by Mizukami into the Bo processor and method because of the benefits of diagnosing particular diseases and/or disease treatments as taught by Mizukami. It also would have been obvious to one of ordinary skill in the art at the time the application was filed to incorporate an alarm such as taught by Bo based on the reticulocyte information because of the ability to detect diseases and other conditions based on the reticulocyte information as taught by Mizukami.
Applicant's arguments filed October 23, 2025 have been fully considered but they are not persuasive. In response to the changes made by applicant, a new drawing objection has been made and the obviousness rejection reference combination has been substantially modified with the addition of a newly cited and applied primary reference, using the previous primary reference as a secondary reference and adding a newly cited and applied secondary reference to the previously applied references.
With respect to the arguments, since they are directed toward Narisada as the primary reference and the current rejection uses the Bo references as the primary reference, the arguments are moot.
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure. The additionally cited art is related to flow cytometers, measurement of various blood particles and composition of diluents and staining solutions used in platelet and/or reticulocyte analysis apparatus.
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/ARLEN SODERQUIST/ Primary Examiner, Art Unit 1797