FINAL OFFICE ACTION
This application has been assigned or remains assigned to Technology Center 1700, Art Unit 1774 and the following will apply for this application:
Please direct all written correspondence with the correct application serial number for this application to Art Unit 1774.
Telephone inquiries regarding this application should be directed to the Electronic Business Center (EBC) at http://www.uspto.gov/ebc/index.html or 1-866-217-9197 or to the Examiner at (571) 272-1139. All official facsimiles should be transmitted to the centralized fax receiving number (571)-273-8300.
Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
Receipt is acknowledged of papers submitted under 35 U.S.C. § 119, which papers have been placed of record in the file.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
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 10 and 11 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of KLEARMAN (US 5376072) and WO 2013/0485546 and OBERLI (US 3578291).
JP 2006-051505 A discloses in Figures 1-14 and per the machine translation reproduced below a method including steps that include providing a sample tube receptacle 30 (Figure 3) with a cylindrical wall and a first end and a second end; wherein the first end is closed by a bottom surface/base 33 to form a receiving space to receive an introduced sample therein; the sample tube receptacle 30 and receiving space therein having a longitudinal axis disposed between the bottom surface/base 33 and the cylindrical wall (at 30); the second end has a closure piece 31 selectively and releasably connected thereto for closing the sample tube receptacle 30 and the sample 74 therein; the closure piece 31 having a lower face directed toward the receiving space presenting a surface facing the receiving space that is capable of inherently producing frictional effects, at least to some degree, as previously argued; wherein an agitator weight element 32 is accommodated within the receiving space and which is shaped and dimensioned such that the agitator weight element 32 can rotate freely in the receiving space only about the longitudinal axis of the receiving space (cf. d, D); the weight element having friction surfaces 34 in the form of structural elements or grooves 34; a device (see Figure 7) for dispersing or homogenizing a sample that comprises a housing 15 with at least one receptacle 22 for a sample tube receptacle 30 which can be converted into a rotational movement (by drive 8);
the method for dispersing or homogenizing a sample 74 further including providing the sample tube receptacle 30 (Fig. 3); introducing the sample 74 into the receiving chamber of the sample tube receptacle 30; the second end has a closure piece 31 that is selectively and releasably connected to said second end for closing the second end of the sample tube receptacle 30 and the sample 74 therein as seen in Figures 3, 6, 11, and 14; the sample 74 can be located above or below the weight element 32 - Figures 3, 6, 11, 14; rotating or turning the sample tube receptacle 30 so that the sample 74 can move below the weight in the direction of gravity - Figures 3, 6, 7, 11, 14; and rotating the sample tube around an axis at 8 - Figure 7.
JP 2006-051505 A:
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Applicant argues that JP 2006-051505 A does not disclose the recited method steps and particular rotational movement specified in claims 10-12. Assuming that this argument has some merit, WO 2013/0485546 discloses steps (a) - (d) of claim 10, particularly described in the underlined and/or bolded wording below:
WO 2013/0485546 thus discloses a multifunctional bioreactor for cell culture and cell sorting, namely bioreactors and more particularly bioreactors for growing and separating cells. Many kinds of cells, especially hematopoietic stem cells and immunocytes, need to be isolated from original samples before they can be efficiently expanded and directed differentiated in culture. This isolation procedure is also called cell sorting or cell separation. Previously, the cell sorting and cell culture have been respectively conducted in separated systems, in which the target cells are isolated first and then are transferred into culture containers. This traditional method is quite cumbersome and has a higher risk for cell contamination and cell loss from the cell sorting to cell culture, in which two completely different devices and systems were involved. Our current invention with a novel design allows these two different procedures completed in one container (chamber), and so minimizes the risk of contamination and target cell loss, and significantly increases the efficiency of the operation.
Some cells are very sensitive to shear-stress in the culture. For example, shear-stress can cause the non-specific differentiation and the increased apoptosis in the stem cell culture, which significantly reduces the efficiency of the stem cell expansion and directed differentiation. The higher shear-stress also causes the more release of non-specific proteins in protein expression, in which the protein of interest takes less proportion in the culture and so result in the increase of protein purification. The static culture has the least shear stress but the cells in static culture normally sit at the bottom of the culture containers, some cells cannot get enough nutrition when cells are at higher density and so not suitable for large scale cell expansion. Some bioreactors, were designed for reducing the shear-stress. However, these bioreactors have to keep cells in suspension by continuously moving, stirring or/and agitating cells. Once the bioreactor stops running, cells will accumulate somewhere of the bottom but are not evenly distributed, which is harmful for most cell growth. Therefore, though the shear-stress has been reduced in these bioreactors, the reduced shear-stress has to continuously exert on the cultured cells when these bioreactors running. In our current invention, when the bioreactor is at static status, cells are allowed to evenly distribute at the bottom of the culture chamber or on the surface of the magnetic beads. Thus, our invention provides cells the best growth condition in both suspension status and static status.
Some bioreactors use magnet element (specifically blades or vans) controlled by magnet impeller to agitate culture media to keep cells in suspension status. This kind of bioreactor purposely enhances the shear-stress for the culture requirements of a certain cells. In addition to the differences in the application, the bioreactor in our invention does not use blades or canes to be the magnet element, and the magnet beads in our invention actually has no magnetism if they are not in magnet field and they can only gain magnetism when they are placed in magnet field. The magnet beads in our invention are not controlled by impeller but by the changes of magnetic field strength affecting the beads' moving. The proper microenvironments, or so called niches, are very important for the growth of some cells, such as stem cells. Some devices use solid materials to form niches or use gel-like materials to form niches. With these devices, after cells culture, cells need to be rinsed out from niches with a special procedure or the niche-forming materials has to be melted or digested with enzyme to release the target cells. In our current invention, the interspaces among beads naturally from the niches for cell growth, and the cells can be easily released when the beads are lifted by changing the magnet field strength. The current invented bioreactor is superior to the welled plate because the mobility of the beads creates dynamic microenvironments for cells needing increased flux of medium to grow. It is also superior to micro-chambers and micro-sieves because it poses less of a challenge to manufacture, is easier to sterilize after use, and the size of the niche can be easily modified by adjusting the size of the beads.
Many cell culture containers (chambers) have been designed for bioreactor use, such as common cell culture flasks, gas-permeable bags, rotation wall vessels, and so on. These containers can be used with common perfusion system with which the cells can be diluted and media can be changed. However, during the media change with these containers, the cytokines, proteins and other expensive substances for cell growth and cells' products are removed from the culture simultaneously. And, the efficiency of common dialysis process is not high enough. The cell culture chambers designed in our current invention take the advantage of colloid osmotic force differences between osmosis chambers at two side of culture chamber to allow the media exchange go through the dialysis membranes rapidly without any loss of cytokines, peptides, proteins and other materials in a certain size. This chamber design provides a novel perfusion strategy and the system with it is called as gradient osmosis perfusion system.
Most bioreactors were designed for culturing either adherent or suspension cells. No bioreactors have been reported to support the growth of partial adherent cells yet. The bioreactor in our current invention can be used for the culture of almost any cell, including suspension cells, adherent cells and partial adherent cells.
It is very common to use computer to control bioreactor running, and many bioreactors are programmable. It is emphasized in our current invention that (1) the strength and direction of magnetic fields, (2)the frequencies and speeds of cell culture chamber flip, and (3) frequencies and speeds of the magnet beads as results of above (1) and (2) are controlled by pre-selected programs or/and programs that response to the data it receives from detectors and send feedback to bioreactor.
Similar to the application of computer in bioreactor control, several cell density detectors with some special light sources (such as laser projector) have been designed for monitoring the cell concentration during the cell culture. The data obtained from the detector or sensor is used to determine if the cells need to be diluted if the culture needs media change. In our current invented bioreactor, the cell density detector uses the common light source and the data is specifically used to adjust the strength of the magnetic field, the flipping speed and frequency of cell culture chamber, as well as indirectly adjust the moving speed and frequency of the magnet beads.
Bioreactors are typically employed to grow cells within a culture. However, many kinds of cells need to be isolated from original samples before they can be efficiently expanded and directed differentiated in culture. This isolation procedure is also called cell sorting or cell separation. Typically, the cell sorting and cell culture have been conducted in separated systems, in which the target cells are isolated first and then are transferred into culture containers. This traditional method is quite cumbersome and has a higher risk for cell contamination and cell loss from the cell sorting to cell culture, in which two completely different devices and systems are involved. Additionally, many bioreactors employ rotating impellers or the like for mixing the contents of the bioreactor. Unfortunately, this imparts a shear stress on the cells which may damage the cells, reduces the efficiency of the system, causes a release of waste products, such as non-specific proteins expression, or the like. Additionally, in some bioreactors, cells may aggregate on the bottom of the cell culture chamber or other locations within the cell culture chamber. The aggregation of these cells in the chamber is not conducive to efficient cell growth. Therefore, there exists a significant need for an efficient bioreactor capable of growing and separating cells therewithin while also minimizing shear-stress imparted to the cells.
In one embodiment, a bioreactor system for growing and separating cells comprises a cell culture chamber comprising an interior having a first portion and a second portion; an agitator element of some weight disposed within the chamber interior, the agitator capable of moving between the interior first portion and interior second portion; and a control system coupled to the cell culture chamber, the control system operable to cause the agitator to move between the interior first portion and interior second portion.
In another embodiment, a method for growing and separating cells comprises providing a chamber comprising an interior having a first portion and a second portion; disposing an agitator within the interior, the agitator capable of moving between the interior first portion and interior second portion; delivering a cell culture media [sample] to the chamber interior; transferring target cells [sample] to the chamber interior; moving the agitator between the first portion and second portion so as to mix the cells and media; and removing waste from the chamber interior while maintaining a substantial number of the target cells within the chamber interior.
Referring now to FIG. 1, a bioreactor system 10 for growing and separating cells is shown. The system 10 includes a cell culture chamber 15, an agitator 20 and a control system 30. The cell culture chamber 15 includes an interior 35 for receiving and growing target cells in a cell culture media [sample] disposed therein, a first end 40 and a second end 45. As used herein, "target cells" refers to cells disposed within the chamber 15 and which are grown within the chamber 15. While the present disclosure is given the context of growing target cells, it will be appreciated that the system may be employed to mix chemicals or any other suitable solution or material. Also, while the first end 40 and second end 45 are shown as being at the top and bottom of the chamber 15 respectively, it will be appreciated that the ends 40, 45 may be in any suitable orientation relative to one another (e.g., in a horizontal plane) and remain within the scope of the present disclosure. As will be discussed below, the chamber 15 may include one or more interior compartments. In addition, as will be appreciated by those skilled in the art, the chamber 15 may be formed from any suitable material, including a rigid material, a flexible material, a combination of rigid and flexible materials, a gas permeable material or any other suitable material. The chamber 15 may further include one or more ports for providing fluid communication between the chamber interior 35 and one or more reservoirs. Illustrative reservoirs include, without limitation, a cell culture media reservoir, a waste reservoir, a buffer reservoir, a C02 reservoir, or any other suitable reservoir.
The agitator 20 is disposed within the chamber interior 35 and is capable of moving between the chamber first end 40 and chamber second end 45. Alternatively, the agitator 20 may be configured to be moved between any two or more points, or between any two or more portions, within the chamber interior 35. In the illustrative embodiment, the agitator comprises a plurality of beads 21. It will be appreciated that any illustrative embodiment showing beads 21 may use any alternative agitator configuration and remain with the scope of the present disclosure and that any particular illustrative embodiment is not limited to using beads exclusively as the agitator. In one embodiment, the beads 21 are be formed from a magnetizable material, such as silicon steel, Fe304, or any other suitable magnetizable material. As used herein, magnetizable means that the agitator, such as the beads, will hold a magnetic charge when subjected to a magnetic field but will not otherwise hold a magnetic charge once removed from the magnetic field, or the magnetic field removed from the vicinity of the agitator, for example, when a magnetic field generator is de-energized. The magnetizable material typically comprises the core of each bead 21. The magnetizable core may then be coated with any suitable material. In one embodiment, the magnetizable core is coated with polystyrene; however, it will be appreciated that the magnetizable core may be coated with any suitable material and remain within the scope of the present disclosure. For example, and without limitation, the magnetizable core may be coated with any suitable thermoplastic or thermoset polymer. While the beads 21 are shown as being formed from a magnetizable material, it will be appreciated that the beads may be formed from any suitable material, magnetizable or non-magnetizable, and remain within the scope of the present disclosure. Additionally, it will be appreciated that the beads 21 may each be coated with any suitable material such that the target cells will adhere to the beads as the cells grow within the chamber 15, yet it will be appreciated that beads not coated with a particular material to which target cells will adhere also remain within the scope of the present disclosure. In some embodiments, it may be desirable to have beads 21 that are buoyant within the cell culture media; therefore, the core of the beads may include air pockets or bubbles, a lightweight foam or plastic or any other suitable material for permitting the beads 21 to be buoyant within the media.
The beads 21 may be formed such that one or more niches, or micro-environments, may be formed or created in the voids between the beads 21 when the beads are stacked together. In some embodiments, these niches may promote growth of additional target cells therein. In one embodiment, where the beads are substantially spherical, the diameter of each bead 21 may be between 1mm and 10mm for the creation of suitable niches. However, it will be appreciated that the beads 21 may have any suitable size and/or shape such that one or more suitable niches may be formed when the beads 21 are stacked together. Also, it will be appreciated that at least some niches may be formed between some beads and one or more walls of the chamber interior.
In an alternative embodiment, as shown in FIG. 5, the agitator 20a may be a plate member 21a having a plurality of apertures 22 therein. The cross-section of the plate member 21a may be complimentary to the cross-sectional shape of the chamber 15 such that the agitator 20a may move within the chamber interior 35. The apertures 22 may permit the media to flow through the agitator 20a as the agitator 20a moves within the chamber interior 35. The agitator 20a may be formed from magnetizable materials or non-magnetizable material, formed to be buoyant or non-buoyant, and/or coated as previously discussed with respect to the beads 20.
Referring again to FIG. 1, the control system 30 may include one or both of a controller 55 and computer 60 for controlling operation of the system 10. Alternatively, the system 10 may be run manually. The control system 30 is configured to be releasably coupled to the chamber 15. The control system 30 may include a cassette 50 for receiving the chamber 15 but it will be appreciated that the chamber 15 may be coupled to the control system 50 via any suitable means or configuration (e.g., clips, hooks, magnets, hook-and-loop assemblies, friction fit, etc.) and remain within the scope of the present disclosure.
The control system 30 may also include a light source 2 and a cell detector 9 for detecting the number of cells within the chamber 15, detecting the change in the number of cells within the chamber 15 or the like and reporting the results back to the control system 30. However, it will be appreciated that any detector, mechanism or technique known in the art for monitoring the number of cells or the change in the number of cells may be employed and remain within the scope of the present disclosure. Additionally, the control system 30, via any suitable detection device, mechanism or method, may monitor the any suitable parameter involved in the growth of the target cells, for example and without limitation, the change in the number of target cells, pH, C02, glucose, calcium, potassium, sodium, temperature, humidity or any other suitable factor and adjust the frequency and/or speed of the movement of the agitator within the chamber and/or adjust the amount of media, the type of media, the amount of buffer, the type of buffer, the amount of C02, or make any other suitable adjustment based on any control system measurement so as to enhance or promote the growth of the target cells within the chamber 15.
The control system 30 is operable to cause the agitator to move within the interior 35 of the chamber 15. This may be accomplished a variety of ways. In the illustrative embodiment, the control system includes a motor 65 operable to rotate the chamber 15 between a first position and second position. As will be discussed below, the first position and second position are approximately 180° apart but it will be appreciated that first and second positions may have any suitable angular relationship relative to one another and remain within the scope of the present disclosure. The chamber 15 may be rotated in a horizontal plane, rotated in a vertical plane or rotated, shifted, slid or otherwise moved in any suitable manner to cause the agitator 20 to move within the chamber 15. In addition, the control system 30 may include first and second magnetic field generators 70, 72 for exciting the beads 21, or other agitator 20, so as to move the beads 21 within the chamber 15 to mix the target cells and culture media. In the illustrative embodiment, each magnetic field generator is an electromagnet that generates a magnetic field when energized and ceases to create a magnetic field when de-energized. When energized, each magnetic field generator draws the agitator 20, e.g. the beads 21, toward the energized magnetic field generator. In an alternative embodiment, a permanent magnet may be used wherein the control system 30 is operable to remove the magnet from the vicinity of the chamber 15 or otherwise block the magnetic field from the magnet from penetrating into the chamber 15. While the illustrative embodiment employs both chamber rotation and electromagnets for moving the agitator within the chamber, it will be appreciated that chamber rotation may be used alone or that electromagnets may be used alone. Moreover, it will be appreciated that any technique for moving the agitator within the chamber may be employed and remain within the scope of the present disclosure.
Referring now to FIG. 2a-2f, operation of the system 10 is illustrated by way of a non-limiting example. Target cells and cell culture media are delivered to the interior 35 of the chamber 15. In this embodiment, the beads 21 are buoyant and float near the top of the cell culture media within the chamber 15. In FIG. 2a, the first magnetic field generator 70 is energized and the beads 21 are held near the end 40 of the chamber 15 [thus the sample is located below the agitator weight element 21 in the vertical plane]. The chamber 15 is then rotated approximately 180° to a position as shown in FIG. 2b wherein the first magnetic field generator 70 maintains the beads 21 near the chamber end 45 [thus the sample is located between the agitator weight element 21 and the top end 40 in the vertical plane]. The first magnetic field generator 70 may then be de-energized whereby the beads 21 begin to float towards the end 40 of the chamber 15 as shown in FIG. 2c. In embodiments where the beads 21 include a coating which target cells will adhere to, movement of the beads 21 from one end to the other will collect newly grown target cells. The target cells may adhere to the beads 21 while waste is flushed from the chamber 15 and/or when new media is introduced to the chamber 15 such that a substantial number of the target cells, original and newly grown, remain within the chamber. Alternatively, magnetizable antibodies specific to the target cells may be added to the interior of chamber 15 whereby the antibodies will bind themselves to the target cells, and when a magnetic field is introduced to the chamber, the antibody bound target cells will be releasably coupled to the magnetizable beads 21 and/or the chamber wall(s) adjacent to the magnetic field generator(s). In this embodiment, one or both of the magnetic field generators 70, 72 may remain energized while unbound cells and/or waste are flushed from the chamber and/or while new media is introduced to the chamber such that a substantial number of the target cells, original and newly grown, remain within the chamber 15. Alternatively, magnetic reagents, such as Annexin V or other suitable reagent, may be employed to couple to damaged or dead cells to the beads and the healthy target cells flushed from the system 10. Further, it will be appreciated that magnetizable antibodies and/or reagents may be employed in a chamber 15 without the use of an agitator whereby the target cells or damaged/dead cells may be held against the chamber when the chamber is flushed.
Once the beads 21 are near the end 45 of the chamber, the second magnetic field generator 72 may be energized whereby the beads 21 are held near the chamber end 40 (FIG 2d) and the chamber rotated to the position shown in FIG. 2e. The second magnetic field generator 72 may then be de- energized whereby the beads 21 will float towards the chamber first end 40 as shown in FIG. 2f. As will be appreciated by those skilled in the art, a variety of additives, media, buffers, C02 and the like may be selectively added to the chamber at any desired point during this process and/or waste selectively removed in order to promote or enhance new cell growth based on measurements taken by the control system as previously discussed.
In an alternative embodiment, non-buoyant beads may be employed such that the beads are moved within the chamber by rotation of the chamber and without also being subjected to magnetic fields. Here, gravity and centrifugal force, by way of rotation of the chamber, are employed to move the beads between two or more points [ends] within the chamber 15 [thus locating the sample in the container between the agitator weight element 21 and one of the ends (one of 40 or 45, e.g., in Figures 1, 2b and 2e) and also locating the sample in the container below the agitator weight element and the other end, e.g., in Figures 2a and 2d upon rotation of the container 15] - see Figures 1 and 2a - 2f reproduced below.
In yet another alternative, the first and second magnetic field generators 70, 72 may be alternately energized so as to move the beads between two or more points within the chamber and without any rotation of the chamber 15. While the foregoing example employs beads 21 as the agitator, it will be appreciated that suitable device may be employed as the agitator and remain within the scope of the present disclosure, including but not limited to that of FIG. 5. Moreover, it will be appreciated that any means or technique for moving the agitator within the chamber may be employed and remain within the scope of the present disclosure.
Accordingly, in view of the teachings and suggestions in WO 2013/0485546, it would have been obvious to one skilled in the art before the effective filing date of the invention to have employed the recited steps that are disclosed WO 2013/0485546 in the analogous method of JP 2006-051505 A for the purpose of rotating the sample tube to therefore move the agitator weight element between the ends of the sample tube receptacle/container and thus through the volume of the sample within the sample tube to enhance mixing, dispersion, and/or homogenization of the contents of the sample tube via rotational/turning movement of the sample tube (i.e., the sample, chemicals, or other solutions as outlined in the description above, see underlined wording above and the Figures).
Modified JP 2006-051505 A thus discloses the recited subject matter as outlined above. Assuming, arguendo, that JP 2006-051505 A does not disclose the recited friction surface on the closure piece, KLEARMAN discloses a sample tube 22 with a closure piece 30 having a frictional surface 32 in the form of structural elements represented as teeth, pyramids, or grooves at 32 (Figures 2, 4, 5, and 8). It would have been obvious before the effective filing date of the invention to have provided the closure piece in modified JP ‘505 with a frictional surface having structural elements as disclosed by KLEARMAN for the purposes of enabling shearing/agitating effects to be produced within the substances present in the tube via said structural elements of the frictional surface.
Modified JP 2006-051505 A thus discloses rotational motion of the sample tube but not the specific oscillating rotational movement about the longitudinal axis of the sample tube recited by claims 10 and 11.
The patent to OBERLI discloses a test tube/sample tube in FIGS. 1 and 2 that is made preferably of an inexpensive impervious material, such as polypropylene for example, and includes a tubular sidewall portion 3 and an integral bottom wall portion 4. The upper surface of the bottom wall is planar and is arranged at a right angle with respect to the longitudinal axis 5. Disposed within the tube 3 are double knife edge stirring means including symmetrical pointed portions 6, 6' that extend diametrically radially outwardly from the longitudinal axis 5 and terminate in longitudinal edges 7, 7' that are adjacent and spaced form the inner wall surface to define narrow gaps 8, 8'. The stirring means is integral with and extends upwardly from the bottom wall 4 in concentrically spaced relation to the tubular wall 3. Preferably, the inner wall surface of the tube 3, the upper surface of the bottom 4 and the lateral surfaces of the stirring means 6, 6' all have completely smooth surfaces. Finally, the tube 3 is provided with guide pins 9 and 9' and 10, 10' that project laterally outwardly from the tube for reception in the tube holder that are driven by conventional oscillatory drive means as shown in FIG. 2. The test tube illustrated in the drawing is intended for a capacity within the milliliter range, the height of the tube 3 being on the order of 5.0 centimeters, for a tube having an inside diameter of approximately 1.3 centimeters. During actual use, the test tube is axially inserted or slid downwardly into the supporting means within which it is held securely against torsion by means of the guide pins 9, 9' and 10, 10'. The holder is oscillated about its longitudinal axis 5 by the oscillatory drive means. In the case of a test tube having the dimensions set forth above, it has been found advantageous to employ an amplitude of the rotary oscillations of approximately .+-.40 radians if the frequency of the rotary oscillations is between 5 and 10 cycles per second. Accordingly, the test tube having a capacity within the milliliter range which is designed preferably for being used only once and consists of an essentially cylindrical tube having an integral bottom wall. In accordance with the primary object of the invention, in order to obtain thorough mixing of the contents of the test tube by merely subjecting the test tube as a whole to oscillatory movement about the longitudinal axis thereof.
It would have been obvious to one skilled in the art before the effective filing date of the invention to have employed the rotational motion of OBERLI in the method of modified JP 2006-051505 A for the purposes of producing oscillations that are imparted to the tube thereby creating considerable relative velocities of the liquid with respect to the tubular to thereby produce a vigorous vortex formation which, in turn, brings about a rapid and thorough stirring or intermixture of the liquids in the sample tube and to prevent the formation of bubbles (col. 2, lines 20-33).
Claims 10 and 11 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of ZURCHER (US 8303914 B2) and WO 2013/0485546 and OBERLI (US 3578291).
Modified JP 2006-051505 A thus discloses the recited subject matter as outlined above. Assuming, arguendo, that modified JP 2006-051505 A does not disclose the recited friction surface on the closure piece, ZURCHER discloses a sample tube 12 with a closure piece 31 having a frictional surface layer 41 on the closure piece in the form of a ceramic material (a well-known abrasive or rough surface capable of providing frictional effects – col. 3, lines 16-21). It would have been obvious before the effective filing date of the invention to have provided the closure piece in modified JP 2006-051505 A with a frictional surface as disclosed by ZURCHER for the purposes of enabling shearing/agitating effects to be produced within the substances present in the tube via said frictional surface.
Claims 10 and 11 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of BROZELL et al. (US 6398051) and WO 2013/0485546 and OBERLI (US 3578291).
Modified JP 2006-051505 A thus discloses the recited subject matter as outlined above. Assuming, arguendo, that modified JP 2006-051505 A does not disclose the recited friction surface on the closure piece, BROZELL et al. discloses a sample tube 22 with a closure piece 30 or 40 having a frictional surface 36, 42, 64, 74, 80, or 92 in the form of structural elements 36, 42, 64, 74, 80, or 92 with teeth at 94. It would have been obvious before the effective filing date of the invention to have provided the closure piece in modified JP 2006-051505 A with a frictional surface having structural elements as disclosed by BROZELL et al. for the purposes of enabling shearing/agitating effects to be produced within the substances present in the tube via said structural elements of the frictional surface.
Claims 10 and 11 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of of LANCESSEUR et al. (US 4805789) and WO 2013/0485546 and OBERLI (US 3578291).
Modified JP 2006-051505 A thus discloses the recited subject matter as outlined above. Assuming, arguendo, that modified JP 2006-051505 A does not disclose the recited friction surface on the closure piece, LANCESSEUR et al. discloses a sample tube 1, 2 with a closure piece 3 having a frictional surface 3a in the form of structural elements 3a. It would have been obvious before the effective filing date of the invention to have provided the closure piece in modified JP 2006-051505 A with a frictional surface having structural elements as disclosed by LANCESSEUR et al. for the purposes of enabling shearing/agitating effects to be produced within the substances present in the tube via said structural elements of the frictional surface.
Claims 10 and 12 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of KLEARMAN (US 5376072) and WO 2013/0485546 and BROUHARD, JR. (US 4184777).
Modified JP 2006-051505 A thus discloses rotational motion of the sample tube but not the intermittent rotational movement about the longitudinal axis of the sample tube recited by claims 10 and 12. The patent to BROUHARD, JR. discloses a liquid agitator which intermittently oscillates a liquid container 44 about an upright axis. A liquid agitator is provided which intermittently oscillates a liquid container about an upright axis and which, at predetermined times, temporarily abruptly stops movement of the container. The agitator keeps the solution in the container properly mixed without constant supervision by intermittently oscillating the container about an upright axis and keeps the solution free of air bubbles and the solution properly mixed without constant supervision by intermittently oscillating the developing tank about an upright axis and periodically imparting a jarring action on the developing tank. An agitator is provided that which includes an upright oscillatable shaft which is spring-biased to a neutral position and is adapted to support a solution container, a motor-driven shaft normal to the upright shaft and carrying a pair of radial arms, and a pair of vertically spaced arms carried by the upright shaft extending across the path of movement of the radial arms.
It would have been obvious to one skilled in the art before the effective filing date of the invention to have employed the rotational motion of BROUHARD, JR. in the method of modified JP 2006-051505 A for the purposes of producing of providing a gentle and intermittent agitation to a solution within a container to keep the solution properly mixed, and, at the same time, imparts an occasional jarring action on the tank to knock free any gas bubbles (col. 2, lines 15-30).
Claims 10 and 12 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of ZURCHER (US 8303914 B2) and WO 2013/0485546 and BROUHARD, JR. (US 4184777).
Modified JP 2006-051505 A thus discloses the recited subject matter as outlined above. Assuming, arguendo, that JP 2006-051505 A does not disclose the recited friction surface on the closure piece, ZURCHER discloses a sample tube 12 with a closure piece 31 having a frictional surface layer 41 on the closure piece in the form of a ceramic material (a well-known abrasive or rough surface capable of providing frictional effects – col. 3, lines 16-21). It would have been obvious before the effective filing date of the invention to have provided the closure piece in modified JP 2006-051505 A with a frictional surface as disclosed by ZURCHER for the purposes of enabling shearing/agitating effects to be produced within the substances present in the tube via said frictional surface.
Claims 10 and 12 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of BROZELL et al. (US 6398051) and WO 2013/0485546 and BROUHARD, JR. (US 4184777).
Modified JP 2006-051505 A thus discloses the recited subject matter as outlined above. Assuming, arguendo, that JP 2006-051505 A does not disclose the recited friction surface on the closure piece, BROZELL et al. discloses a sample tube 22 with a closure piece 30 or 40 having a frictional surface 36, 42, 64, 74, 80, or 92 in the form of structural elements 36, 42, 64, 74, 80, or 92 with teeth at 94. It would have been obvious before the effective filing date of the invention to have provided the closure piece in modified JP 2006-051505 A with a frictional surface having structural elements as disclosed by BROZELL et al. for the purposes of enabling shearing/agitating effects to be produced within the substances present in the tube via said structural elements of the frictional surface.
Claims 10 and 12 are rejected under 35 U.S.C. 103 as being unpatentable over JP 2006-051505 A in view of LANCESSEUR et al. (US 4805789) and WO 2013/0485546 and BROUHARD, JR. (US 4184777).
Modified JP 2006-051505 A thus discloses the recited subject matter as outlined above. Assuming, arguendo, that JP 2006-051505 A does not disclose the recited friction surface on the closure piece, LANCESSEUR et al. discloses a sample tube 1, 2 with a closure piece 3 having a frictional surface 3a in the form of structural elements 3a. It would have been obvious before the effective filing date of the invention to have provided the closure piece in JP ‘505 with a frictional surface having structural elements as disclosed by LANCESSEUR et al. for the purposes of enabling shearing/agitating effects to be produced within the substances present in the tube via said structural elements of the frictional surface.
Allowable Subject Matter
None.
Response to Amendment
Applicant's response and arguments filed 19 MAY 2026 including no revisions to the claims have been considered but they are not persuasive.
Both JP 2006-051505 A and WO 2013/048546 disclose introducing a sample into a sample tube receptacle. From the available machine translation of JP 2006-051505 A reproduced below, the term “sample” appears, in bold, 67 times:
SAMPLE CRUSHING IMPLEMENT
Patent Number: 2006051505
Document ID: JP 2006051505 A
Date Published: 2006-02-23
Abstract
<P>PROBLEM TO BE SOLVED: To provide a sample crushing implement efficiently crushing even a large or hard sample. <P>SOLUTION: This sample crushing implement is provided with a thin and long bottomed cylindrical vessel 30 having a vessel body with a truncated conical recessed bottom, and a lid 31 openably/closably sealing its opening; and a crushing medium 32 contained in the vessel 30, with its body part columnarly formed so that the body part keeps inside the vessel body an attitude approximately along its axis, and relatively moves in the axial direction, with its one end truncated and conically protruded to correspond to the bottom shape of the vessel body and continuous to the body part. <P>COPYRIGHT: (C)2006,JPO&NCIPI
Description
TECHNICAL-FIELD
The present invention relates to a sample crushing tool used for crushing a sample of plant tissue, seeds, animal tissue, plastic material, mineral material or the like for chemical analysis and fractional separation.
BACKGROUND-ART
In order to chemically analyze and fractionate various samples as described above, the sample must first be uniformly pulverized. Conventionally, crushing using a mortar and pestle is known, but since it is not suitable for efficiently crushing many samples, it is not suitable for crushing containers containing samples and crushing media. A crushing apparatus that crushes many at the same time is used.
In order to crush a sample using the crushing apparatus, a crushing medium is stored together with the sample in a crushing container such as a centrifuge tube that is generally used for centrifuging the sample, and the crushing container is used as a crushing apparatus. Wear and apply vibration to the crushing container. As the crushing medium, fine beads such as glass, ceramic, and metal are known (see Patent Document 1). When vibration is applied to the crushing container by the crushing device, the crushing medium collides with the sample or is pressed against the wall surface of the crushing container and is crushed. Therefore, the sample is crushed by applying vibration for the required time to the crushing container.
A frozen sample crushing cell is known as an instrument for crushing a sample (see Patent Document 2). This is done by freezing the sample in order to make it easy to crush the sample, putting it into a cooled bottomed cylindrical container, and installing a lid with a protruding part having a shape corresponding to the inner shape of the container. The sample is inserted into a container and placed on the container, and the frozen sample is crushed by applying a vibration press to the lid.
[1] Japanese Examined Patent Publication No. 6-036732 (pages 2 and 3, FIG. 1) [2] JP-A-8-219957 (pages 1 and 2, FIG. 1)
DISCLOSURE
TECH-PROBLEM
However, in the crushing method using microbeads as a crushing medium, when the sample becomes a large plant cell, animal cell, mineral material, etc., there is a problem that the sample is difficult to move and the crushing medium is not crushed because the mass of the crushing medium is small. there were.
In addition, in order to use the crushed sample for processing such as centrifugation and analysis, the frozen sample crushing cell needs to transfer the crushed sample from the container to another container necessary for the processing. If the sample contains bacteria or the like, it may be dissipated. In addition, there is a problem that operations such as washing and sterilization are necessary for use in the next crushing.
The present invention has been made in view of the above-described conventional problems, and an object thereof is to provide a sample crushing tool capable of efficiently crushing large plant tissues, animal tissues, mineral materials, and the like.
TECH-SOLUTION
The first invention of the present application for achieving the above object is a sample crushing tool for storing a crushing medium together with a sample to be crushed in a crushing container, and for crushing the sample by vibrating the crushing container. A crushing container having a bottom cylindrical shape, and a bottom body having a substantially hemispherical shape, a semi-elliptical sphere shape or a conical conical shape, and a capping container that seals its opening so as to be openable and closable, and the crushing vessel The main body is formed in a cylindrical shape so as to move in the axial direction while maintaining a posture substantially along the axis of the container main body, and one end of the main body of the container main body. And a crushing medium having a substantially hemispherical shape, a semi-elliptical spherical shape or a truncated cone shape corresponding to the shape of the bottom surface and projecting so as to be continuous with the main body portion.
When the crushing medium is accommodated together with the sample in the crushing container, and the reciprocating vibration is applied to the crushing medium in a direction substantially along the axis thereof, preferably the reciprocating vibration in the shape of 8 is applied, Since the operation of colliding with the bottom surface is repeated, the crushing container acts like a mortar and the crushing medium acts like a pestle so that the sample is effectively crushed. Even if the sample is a large plant tissue, animal tissue, plastic material or mineral material, it can be efficiently crushed.
The second invention of the present application is a sample crushing tool for storing a crushing medium together with a sample to be crushed in a crushing container, and for crushing the sample by vibrating the crushing container. The container body recessed in a substantially hemispherical shape, a semi-elliptical spherical shape, or a truncated cone shape, and its opening are sealed so that it can be opened and closed, and the ceiling surface is recessed in a substantially hemispherical shape, a semi-elliptical spherical shape or a truncated cone shape. A crushing container provided with a lid, and a circular body so that the main body is accommodated in the crushing container and the main body portion is relatively moved in the axial direction while maintaining a posture substantially along the axis in the container main body. It is formed in a columnar shape, and has one end portion that is substantially hemispherical, semi-elliptical spherical, or conical so as to correspond to the shape of the bottom surface of the container main body, and is provided so as to be continuous with the main body portion. Almost hemispherical, semi-elliptical so that the end corresponds to the ceiling surface shape of the lid A Jo or frusto conical and so as to continuously to the main body portion and a convexly been crushed medium, characterized by comprising. According to this, since the sample can be crushed on both the bottom surface and the ceiling surface of the crushing container, more effective crushing can be performed.
In the above configuration, when the difference between the inner diameter of the crushing container and the outer diameter of the crushing medium is 2 mm or less, even when the sample is supple, the crushing medium can be acted reliably and can be efficiently crushed.
Further, by forming one or a plurality of grooves on the surface of at least one end of the crushing medium, the fiber is effective in the grooves even in the case of animal tissues or plant tissues that contain fibers that are difficult to cut. Disconnected.
In addition, if the sample is frozen in a liquid nitrogen bath and freeze-dried, even if the sample contains fibers that are supple and difficult to cut, the sample will be easily cut by freezing, and the sample will be crushed. Is made efficient.
In addition, if the crushing medium is made of titanium whose surface is polished, the sample can be crushed even in animal tissues such as bone, and tissue such as DNA and RNA remains on the surface of the crushing medium, This eliminates the possibility of cross contamination occurring during crushing.
ADVANTAGEOUS-EFFECTS
As described above, according to the sample crushing tool according to the present invention, the crushing medium is housed together with the sample in the crushing container, and the crushing medium is rotated relative to the bottom surface of the crushing container by reciprocating vibration. Or since it collides repeatedly with a bottom face and a ceiling surface, a crushing container acts like a mortar and a crushing medium acts like a pestle, and a sample is crushed effectively. Even if the sample is a large plant tissue, animal tissue, plastic material, mineral material, etc., it can be efficiently crushed.
BEST-MODE
Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. The present embodiment is an example embodying the present invention, and does not limit the technical scope of the present invention.
1 and 2 show configurations of the crushing container 30 and the crushing medium 32 according to the first embodiment. The crushing container 30 is formed of an elongated cylindrical container, and a screw 30a is formed on the outer periphery of the opening, and a mount cone part 33 is formed on the bottom, so that the lid 31 can be screwed into the opening and sealed. It is configured. An annular seal portion 31 a that fits to the inner periphery of the opening of the crushing container 30 is formed on the inner periphery of the lid 31. The crushing container 30 has a capacity of 2 ml to 50 ml depending on the material and amount of the sample.
The crushing medium 32 accommodated together with the sample in the crushing container 30 is composed of a single member having a length L larger than the inner diameter D of the crushing container 30 as shown in FIG. Corresponding to the bottom shape of the crushing container 30, a similar conical projecting end 32 a is formed in the part. The other end portion is formed in the small diameter portion 32b so as not to interfere with or fit into the annular seal portion 31a on the inner periphery of the lid body 32. Further, the outer diameter d of the crushing medium 32 is set to be 2 mm or less with respect to the inner diameter D of the crushing container 30, and is set to about 1 mm or less when the inner diameter D of the crushing container 30 is small. For example, when the capacity of the crushing container 30 is 2 ml and the inner diameter D is 8 mm, the outer diameter d of the crushing medium 32 is set to 7 mm.
Further, as shown in FIGS. 2B and 2C, the protruding end 32a of the crushing medium 32 may be formed with one or a plurality of grooves 34 formed radially or spirally as necessary. .
4 and 5 show a crushing container 30 and a crushing medium 32 according to the second embodiment. The crushing container 30 shown in FIG. 4 has a hemispherical or semi-elliptical spherical hemispherical portion 35 formed at the bottom thereof, and accordingly, at one end of the crushing medium 32, as shown in FIG. A protruding end portion 32a is formed. Further, as shown in FIGS. 5B and 5C, a groove 34 can be formed in the protruding end portion 32a.
As the material of the crushing container 30 shown in the first and second embodiments, synthetic resins such as polycarbonate, polypropylene, polyethylene, polystyrene, and polyfluorinated ethylene can be used, and there is no need to add a colorant. Therefore, the resin is molded to be transparent or translucent. When the crushing container 30 is transparent or translucent, it is suitable for the case where crushing is performed while checking the state of the sample accommodated in the crushing container 30, particularly the state of progress of crushing.
For example, a screw mouth type centrifuge tube manufactured by Sarstedt (SARSTED: Germany), a Safelock tube manufactured by Eppendorf (Eppendorf: Germany), a centrifuge tube manufactured by Norgen (NALGEN: USA), and the like.
Further, the crushing container 30 may be made of a metal such as stainless steel or a polyfluorinated ethylene coating on the inner surface thereof.
As the material of the crushing medium 32, magnetic stainless steel such as SUS430, stainless steel such as SUS304 hard stainless steel, carbon steel, titanium, tungsten, zirconia, tungsten carbide, ceramic, glass, polyfluoroethylene, etc. are used alone or in combination. What was comprised can be used suitably.
When the sample is crushed using the crushing container 30 and the crushing medium 32 shown in each of the first and second embodiments, as shown in FIGS. The crushing medium 32 and the sample 74 having a longer length are accommodated, the opening of the crushing container 30 is closed with the lid 31, and vibration is applied to the crushing container 30.
The vibration applied to the crushing container 30 is preferably applied so that the crushing medium 32 repeats collision with the bottom of the crushing container 30 while relatively rotating while maintaining the posture substantially along the axis of the crushing container 30. As a means for realizing the above, when the crushing container 30 is attached to the crushing apparatus and a figure-shaped vibration is applied, the crushing process can be efficiently performed on a large number of crushing containers 30 simultaneously.
FIG. 7 shows a configuration of a main part of a crushing apparatus capable of applying an 8-shaped vibration to a large number of crushing containers 30. An inclined shaft body 11 is fixed to a rotary shaft 8 that is rotationally driven by a driving means (not shown), and an annular body 15 is attached to the outer periphery of the inclined shaft body 11 via a pair of bearings 14 so as to be relatively rotatable. Yes. A magnet 16 is attached to an appropriate position at the lower periphery of the annular body 15. Further, a counter electrode magnet 18 supported by a mounting bracket 17 attached to the upper portion of the bearing portion 7 is fixedly disposed so as to face the magnet 16. The attractive force of the magnet 16 and the counter electrode 18 prevents the rotation of the annular body 15 when the rotating shaft 8 and the inclined shaft body 11 rotate, and the annular body 15 swings as the inclined shaft body 11 rotates. Is configured to do.
An attachment step portion 19 is provided on the upper portion of the annular body 15, and an annular holding body 20 is placed thereon and fastened and fixed by a fixing bolt 21. As shown in a plan view in FIG. 8, a large number of elongated container cases 22 are arranged on the outer periphery of the annular holder 20 in a posture parallel to the axis of the annular body 15, and the crushing container 30 is placed in the container case 22. It is configured to accommodate and support. Further, the lid 31 of the crushing container 30 is engaged with the upper end of the container case 22, and a plurality of pressing plates 23 for pressing the upper surface of the lid 31 to fix the crushing container 30 are provided on the annular holding body 20. It is configured to be installed on the provided mounting boss 24 and fastened and fixed with a fixing screw 25. The height of the mounting boss 24 is set to be substantially the same height as the upper surface height position of the lid 31.
When the crushing container 30 is accommodated in the container case 22 of the annular holder 20 and fixed by the pressing plate 23 in the crushing apparatus having the above-described configuration, and the rotary shaft 8 is driven to rotate by a driving means (not shown), the crushing container 30 is moved to its axis. A reciprocating vibration is formed in an 8-shape that combines a main reciprocating movement of a relatively long stroke in the core direction and a sub-reciprocating movement of a relatively short stroke in the direction orthogonal thereto, and the crushing medium 32 is accompanied by the axial core of the crushing container 30. The crushing container 30 acts like a mortar, the crushing medium 32 acts like a pestle, and the object to be crushed is a large plant cell or animal. Even tissue, plastic materials and mineral materials are efficiently crushed.
The crushing container 30 and the crushing medium 32 can be configured as shown below in addition to the configurations shown in the first and second embodiments.
9 and 10 show a crushing container 30 and a crushing medium 32 according to the third embodiment. The crushing container 30 shown in FIG. 9 has a head cone portion 33 formed on both the bottom and the lid 31, and accordingly, the crushing medium 32 is the same at both ends as shown in FIG. 10 (a). The projecting end 32a having a conical shape is formed. Further, as shown in FIGS. 10B and 10C, one or a plurality of radial or spiral grooves 34 can be formed over the entire length of the protruding end portion 32a and the crushing medium 32 as necessary.
12 and 13 show a crushing container 30 and a crushing medium 32 according to the fourth embodiment. The crushing container 30 shown in FIG. 12 has a hemispherical portion 35 formed on both the bottom portion and the lid 31, and a similar hemispherical protruding end portion 32 a is formed on both ends of the crushing medium 32. Further, as shown in FIGS. 13B and 13C, one or a plurality of radial or spiral grooves 34 can be formed over the entire length of the protruding end 32a and the crushing medium 32 as necessary.
As shown in FIGS. 9 and 12, both end portions of the crushing container 30 including the lid body 31 are the head cone portion 33, the hemispherical portion 35, the semi-elliptical sphere portion, etc. When the shape is substantially the same, the mortar-pestle action can be obtained more effectively, and more efficient crushing can be achieved.
In the crushing of the sample by the crushing container 30 and the crushing medium 32 described above, if the difference between the inner diameter of the crushing container 30 and the outer diameter of the crushing medium 32 is 2 to 1 mm or less, the crushing medium can be obtained even when the object to be crushed is supple. 32 can be made to act reliably and can be efficiently crushed.
In addition, in the case of animal and plant tissues that contain fibers that are pliable and difficult to cut, the crushed object is frozen in a liquid nitrogen bath with the crushed container 30 accommodated in the crushing container 30. However, when freeze-drying, the material to be crushed is easily crushed, so that it can be efficiently crushed. In the present invention, the same effect can be obtained only by cooling with dry ice. Furthermore, by cooling the crushing medium 32 together with the object to be crushed with liquid nitrogen or the like, heat generation during crushing can be prevented, and if the crushing medium 32 is cooled to a lower temperature than the object to be crushed, the heat generation is prevented. The effect is remarkable.
Further, when the object to be crushed is a similar animal or plant tissue, grooves 34 are provided as shown in FIGS. 2 and 4 (b) and (c), and FIGS. 10 and 13 (b) to (e). When the crushing medium 32 is used, the fiber that is difficult to be cut by the groove 34 is effectively cut without being freeze-vacuum dried as described above, so that the crushing medium 32 can be efficiently crushed. In this case, a significant effect is exhibited by cooling with dry ice as described above.
In addition, when a material made of titanium whose surface is polished is used as the crushing medium 32, a composition such as DNA or RNA is present on the surface of the crushing medium 32 even when the object to be crushed is a hard animal tissue such as bone. It is possible to eliminate the risk of cross contamination occurring during the next crushing.
In addition, by using a material obtained by coating the surface of a metal material such as carbon steel with polyfluorinated ethylene as the crushing medium 32, the crushing container 30 has a corrosion resistance when containing a corrosive buffer solution or an extract. It is possible to secure the necessary weight for the action.
Explanation of symbols
30 Crushing container 31 Lid 32 Crushing medium 33 Mounted cone part 34 Groove 35 Hemisphere part
DRAWING DESCRIPTION
[1]Sectional drawing which shows the structure of the crushing container which concerns on 1st Embodiment. [2]The side view which shows the structure of the crushing medium which concerns on 1st Embodiment. [3]Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container. [4]Sectional drawing which shows the structure of the crushing container which concerns on 2nd Embodiment. [5]The side view which shows the structure of the crushing medium which concerns on 2nd Embodiment. [6]Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container. [7]The principal part sectional drawing which shows the structure of a crushing apparatus. [8]The 1/2 top view which shows the structure of a cyclic | annular holding body. [9]Sectional drawing which shows the structure of the crushing container which concerns on 3rd Embodiment. [10]The side view which shows the structure of the crushing medium which concerns on 3rd Embodiment. [11]Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container. [12]Sectional drawing which shows the structure of the crushing container which concerns on 4th Embodiment. [13]The side view which shows the structure of the crushing medium which concerns on 4th Embodiment. [14]Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container.
Claims
In the sample crushing tool for storing the crushing medium together with the sample to be crushed in the crushing container, and for crushing the sample by vibrating the crushing container,
An elongated bottomed cylindrical shape, a container body with a bottom surface recessed in a substantially hemispherical shape, a semi-elliptical spherical shape, or a truncated cone shape, and a crushing container provided with a lid that seals its opening so that it can be opened and closed;
It is accommodated in the crushing container, and the main body is formed in a cylindrical shape so as to move in the axial direction while maintaining a posture substantially along the axial center in the container main body. A crushing medium projecting so as to be substantially hemispherical, semi-elliptical sphere or conical cone and corresponding to the main body so as to correspond to the bottom shape of the container main body,
A sample crusher characterized by comprising.
In the sample crushing tool for storing the crushing medium together with the sample to be crushed in the crushing container, and for crushing the sample by vibrating the crushing container,
An elongated cylindrical body with a bottom and a bottom that is approximately hemispherical, semi-elliptical, or conical, and its opening is sealed so that it can be opened and closed, and the ceiling is semi-spherical, semi-elliptical A crushing container having a lid recessed in a spherical shape or a truncated cone;
It is accommodated in the crushing container, and the main body is formed in a cylindrical shape so as to move in the axial direction while maintaining a posture substantially along the axial center in the container main body. It is substantially hemispherical, semi-elliptical sphere, or conical so as to correspond to the bottom shape of the container body, and is projected so as to be continuous with the main body, and the other end is formed into the ceiling surface shape of the lid. A crushing medium that is substantially hemispherical, semi-elliptical spherical, or conical in a corresponding manner and is provided so as to be continuous with the main body,
A sample crusher characterized by comprising.
The sample crushing tool according to claim 1 or 2, wherein a difference between the inner diameter of the crushing container and the outer diameter of the main body of the crushing medium is 2 mm or less.
The sample crushing tool according to any one of claims 1 to 3, wherein one or a plurality of grooves are formed on an end surface of the crushing medium.
The sample crushing tool according to any one of claims 1 to 4, wherein the sample is frozen in a liquid nitrogen bath and freeze-dried.
The sample crushing tool according to any one of claims 1 to 5, wherein the crushing medium is made of titanium whose surface is polished.
Accordingly, the base reference to JP 2006-051505 A clearly teaches employing a sample in the sample tube receptacle and any argument to the contrary is misplaced.
On this same issue, WO 2013/048546 also clearly teaches employing a sample in the sample tube receptacle. The explanation of WO 2013/048546 above is replete with term “cells” and “cell culture media” and this reference teaches that the invention thereof can be used for the culture of almost any cell, including suspension cells, adherent cells and partial adherent cells, i.e., reasonably equivalent to a “sample” within the broad expanse of that term (see page 12 above). Said sample is introduced into the sample receptacle 15 (otherwise one wonders in what manner it could be present in the receptacle). Thus, contrary to the remarks, WO 2013/048546 discloses the introduction of the broadly recited “sample” into a sample tube receptacle.
The base reference to JP 2006-051505 A shows the sample 74 located above or below the weight element 32 as depicted in Figures 3, 6, 11, 14 and the sample is rotated or turned by the apparatus seen in Figure 7.
Moreover, WO 2013/048546 shows and describes the receptacle 15 being turned/rotated such that the sample is located between the weight element/agitator beads 21 and the second [bottom] end of the receptacle 15 as seen in Figures 2a and 2d reproduced below:
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Figure 5 represents an embodiment 21a of the agitator that is not specifically relied upon since the claimed weight element, other than including a friction surface, is of a scope to encompass nearly any element possessing a mass, certainly encompassing the embodiment 21 of the agitator beads of WO 2013/048546 seen in the other Figures. The subject matter of Figure 5 is not directly relevant to the rejection and is only included for completeness.
Contrary to the implications made in the remarks, it is noted that the pending claims do not impose a specific order on the performance of the method steps. Altiris Inc. v. Symantec Corp., 318 F.3d 1363, 1371, 65 USPQ2d 1865, 1869-70 (Fed. Cir. 2003) (Although the specification discussed only a single embodiment, the court held that it was improper to read a specific order of steps into method claims where, as a matter of logic or grammar, the language of the method claims did not impose a specific order on the performance of the method steps, and the specification did not directly or implicitly require a particular order). MPEP 2111.01.
In the last line of page 16, WO 2013/048546 discloses that the “chamber 15 may be rotated in a horizontal plane, rotated in a vertical plane or rotated, shifted, slid or otherwise moved in any suitable manner to cause the agitator 20 to move within the chamber [receptacle] 15” which certainly suggests that other rotational protocols for a receptacle, such as those of OBERLI or BROUHARD, JR. or otherwise, may be employed in the rotational movement method for the receptacle 15 of WO 2013/048546.
The remarks then discuss the disclosures of the other references used in the rejections, however, the remarks expose subject matter that is not relied upon in the rejections (such as but not limited to the sharp blades of the stirring element in OBERLI; the drive elements such as motor, gearbox, striking arms, cam rods, etc. in BROUHARD; the rings and snap lock of ZURCHER; spring loaded device of BROZELL; flexible tongue of LANCESSEUR; etc.). Said subject matter is irrelevant to the essence of the rejections since OBERLI and BROUHARD, JR. are simply relied upon for rotational protocols of a receptacle that provide rotational movement about a longitudinal axis of the receptacle, oscillating rotational movement, and intermittent rotations, as recited in claims 10-12. Thus, these references are NOT utilized for all the extraneous subject matter outlined by Applicant in the remarks.
Likewise, the references to ZURCHER, BROZELL, and LANCESSEUR are merely relied upon for the teachings of the disclosed frictional surface(s) and a buttress against any future newly filed claims that may recite such surfaces. Again, these three references are not relied upon for all the superfluous subject matter outlined by Applicant in the remarks.
Such remarks are clearly a typical case of attacking the prior art references individually in a piecemeal fashion. "Non-obviousness cannot be established by attacking references individually where the rejection is based upon the teachings of a combination of references," In re Merck & Co., Inc., 800 F.2d 1091, 1097 (Fed. Cir. 1986). In response to these arguments against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981).
Moreover, all the recited subject matter of the cancelled apparatus claims (such as the particular structural elements, the grooves, kit, weight element, materials, etc.) are indeed known in the art as expressed in prior office actions. Any revised or new claims reciting such subject matter will be rejected as in prior office actions since such subject matter merely embodies knowledge well-known and obvious to one skilled in the art particularly with respect to the frictional surfaces disposed in a sample tube or receptacle as taught by the ZURCHER, BROZELL, and LANCESSEUR references.
With respect to the applied references herein, the examiner has considered all of the disclosure of each reference for what it would have fairly taught one of ordinary skill in the art. In re Boe, 355 F.2d 961, 148 USPQ 507 (CCPA 1966). Additionally, the specific teachings of each reference and the inferences which one skilled in the art would have reasonably been expected to draw from the disclosure has been taken into account. In re Preda, 401 F.2d 825, 159 USPQ (CCPA 1968). On the basis of the knowledge and level of skill in the art at the time of applicant's invention, as reflected by the newly applied references obtained after performing a comprehensive search, the examiner concludes that the rejections under 35 USC 103 are well founded.
Applying the test for obviousness set forth in In re Keller, supra, which is what the combined teachings of the references would have suggested to those of ordinary skill in the art, the examiner concludes that one having ordinary skill in the art would have found it prima facie obvious to have combined the references as set forth in the rejections herein.
With respect to the concept that the prior art must contain something to suggest the desirability of the combination, it is noted that to justify combining reference teachings in support of a rejection under 35 USC 103, it is not necessary that a device shown in one reference be capable of being physically inserted into the device shown in the other or that the prior art suggest expressly the changes or possible improvements the applicant has made. It is only necessary that knowledge clearly present in the prior art was applied. In re Keller, supra; In re Sernaker, 702 F.2d 989, 217 USPQ 1 (Fed. Cir. 1983). The examiner has applied only knowledge clearly present in the prior art as evidenced by the prior art above in the rejections of the pending claims and the rejections are thus proper.
Accordingly, the Examiner has established that, prima facie, one of ordinary skill in this art routinely following the combined teachings of the prior art herein would have reasonably arrived at the claimed method, including each and every limitation thereof arranged as required therein, without recourse to Appellants' Specification. See, e.g., KSR Int'l Co. v. Teleflex Inc., 127 S.Ct. 1727, 1739 (2007) (a patent claiming a combination of elements known in the prior art is obvious if the improvement is no more than the predictable use of the prior art elements according to their established functions); In re Kahn, 441 F.3d 977, 985-88 (Fed. Cir. 2006); In re Keller, 642 F.2d 413,425 (CCPA 1981) (("[T]he test [for obviousness] is what the combined teachings of the references would have suggested to those of ordinary skill in the art."); In re Sovish, 769 F.2d 738, 743 (Fed. Cir. 1985) (skill is presumed on the part of one of ordinary skill in the art). The examiner thus concludes pending claims 10-12 are obvious over the four corners of the prior art references.
Conclusion
Accordingly, THIS ACTION IS MADE FINAL.
Applicant is reminded of the extension of time policy as set forth in 37 C.F.R. § 1.136(a). A SHORTENED STATUTORY PERIOD FOR RESPONSE TO THIS FINAL ACTION IS SET TO EXPIRE THREE MONTHS FROM THE DATE OF THIS ACTION. IN THE EVENT A FIRST RESPONSE 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 EXTENSION FEE PURSUANT TO 37 C.F.R. § 1.136(a) WILL BE CALCULATED FROM THE MAILING DATE OF THE ADVISORY ACTION. IN NO EVENT WILL THE STATUTORY PERIOD FOR RESPONSE EXPIRE LATER THAN SIX MONTHS FROM THE DATE OF THIS FINAL ACTION. ANY RESPONSE FILED AFTER THE MAILING DATE OF THIS FINAL REJECTION WILL BE SUBJECT TO THE PROVISIONS OF MPEP 714.12 AND 714.13 - NO EXCEPTIONS.
Per Rule 1.116(b)(3): “An amendment touching the merits of the application or patent under reexamination may be admitted upon a showing of good and sufficient reasons why the amendment is necessary and was not earlier presented.” Thus, an amendment after final lacking such showing will be denied entry.
Current USPTO policy limits time for interviews to one per new application or RCE (utility), when during prosecution, the examiner conducts an interview. More than one interview and additional time will only be granted if it is ensured “that the interviews are being used to advance prosecution”. Applicant has undoubtedly consumed their allotment of interviews (two as seen below and at least one with supervisory personnel), so no interview will be granted after final.
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Any inquiry concerning this communication or earlier communications from the examiner should be directed to CHARLES COOLEY whose telephone number is (571)272-1139. The examiner can normally be reached M-F 9:30 AM - 6:00 PM.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, CLAIRE X. WANG can be reached at, yet again, at (571)270-1051 (office direct). The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/CHARLES COOLEY/Examiner, Art Unit 1774
9 JUNE 2026
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