CN114015552B - Kit for identifying human mesenchymal stem cells - Google Patents
Kit for identifying human mesenchymal stem cells Download PDFInfo
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- CN114015552B CN114015552B CN202111301926.0A CN202111301926A CN114015552B CN 114015552 B CN114015552 B CN 114015552B CN 202111301926 A CN202111301926 A CN 202111301926A CN 114015552 B CN114015552 B CN 114015552B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
Abstract
The invention discloses a kit for identifying human mesenchymal stem cells, which comprises a basal disc, wherein a reaction groove is formed in the middle of the upper surface of the basal disc, three mounting grooves are formed in the upper surface of the basal disc and positioned at intervals on the outer side of the reaction groove, a reaction agent placing soft bag with a U-shaped section is filled in the part, positioned on the upper surface of the basal disc, of one end of each mounting groove, a sealing ring is arranged in the middle of the upper surface of the basal disc, a connecting channel is formed in the lower surface of the sealing ring corresponding to the upper end of the reaction agent placing soft bag, and the other end of the connecting channel extends to the lower surface of the sealing ring along the ring body of the sealing ring and faces the reaction groove. The method avoids the phenomenon of easy pollution in the traditional liquid material adding process through a dropper in the identification process, greatly improves the success rate of the identification process aiming at the osteogenesis or chondrogenesis differentiation direction of the human mesenchymal stem cells, is simple and convenient to operate, and is beneficial to popularization and application.
Description
Technical Field
The invention relates to the technical field of cell identification, in particular to a kit for identifying human mesenchymal stem cells.
Background
Bone marrow tissue contains many cellular components, including two types of pluripotent stem cells in addition to differentiated cells such as stromal cells: hematopoietic stem cells and mesenchymal stem cells are commonly identified in medical science aiming at the mesenchymal stem cells, in particular to human mesenchymal stem cells.
In the process of identifying the human bone marrow mesenchymal stem cells, the identification of the multidirectional differentiation potential is included, and in the process of identifying the multidirectional differentiation potential, the differentiation in the bone or cartilage direction is required to be carried out: firstly, digesting (removing culture solution of 3 rd generation logarithmic growth phase bone marrow mesenchymal stem cells, washing twice by PBS, digesting for a plurality of minutes by 0.25% pancreatin solution, adding 10% FBS DMEM culture medium to stop digestion, and centrifuging for a plurality of minutes at 1500 r/min); ② inducing an L-DMEM culture medium (adding a mesenchymal stem cell inducing liquid into the centrifuged liquid).
In the identification process, the liquid material adding process is complicated, and the steps are various, so that the condition that the liquid material is polluted by the external environment is easy to occur in the adding process, the phenomenon of identification failure is easy to occur, and the success rate of identification is reduced. To this end, we propose a kit for identifying human mesenchymal stem cells.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a kit for identifying human mesenchymal stem cells.
In order to achieve the purpose, the invention adopts the following technical scheme:
a kit for identifying human mesenchymal stem cells comprises a basal disc, wherein a reaction groove is formed in the middle of the upper surface of the basal disc, three mounting grooves are formed in the upper surface of the basal disc and in the outer side of the reaction groove at intervals, a reaction agent placing soft bag with a U-shaped section is filled in a part, located at one end of each mounting groove, of the upper surface of the basal disc, the upper end of the reaction agent placing soft bag is open and is flush with the upper surface of the basal disc, the other end of each mounting groove extends to the outer ring surface of the basal disc, an internal thread ring is embedded in a part, close to the outer ring surface of the basal disc, of each mounting groove, an extrusion magnet rod penetrates through the internal thread ring, one end of each extrusion magnet rod extends to the outer ring surface of the basal disc, and an extrusion iron ball is arranged between one end, located in the basal disc, of each extrusion magnet rod and the reaction agent placing soft bag;
the middle part of the upper surface of the base plate is provided with a sealing ring, the inner ring surface of the sealing ring extends to the inner side of the ring groove surface where the reaction groove is located, the lower surface of the sealing ring covers the upper end of the mounting groove and is close to the outer side of the upper surface of the base plate, the lower surface of the sealing ring corresponding to the upper end of the reactant placing soft bag is provided with a connecting channel, and the other end of the connecting channel extends to the lower surface of the sealing ring along the ring body of the sealing ring and faces the reaction groove;
the three reactants are respectively placed in the soft bags: removing culture solution from the 3 rd generation logarithmic growth phase bone marrow mesenchymal stem cells, washing with PBS for 2 times, collecting the residual solution, pancreatin solution with concentration of 0.25%, and inducing solution of bone marrow mesenchymal stem cells.
Preferably, the diameter length of the inner ring of the sealing ring is three quarters of the diameter of the upper end of the reaction tank.
Preferably, the upper surface of sealing ring is equipped with the joint ring, the sealing ring has sealed lid through joint ring card lid, the lower surface of sealed lid closely laminates with the upper surface of sealing ring.
Preferably, the outer side of the upper surface of the base disc is provided with a threaded connection ring corresponding to the outer annular surface of the sealing ring, the inner annular surface of the threaded connection ring is in threaded connection with the outer annular surface of the sealing ring, and the upper surface of the base disc is tightly attached to the lower surface of the sealing ring.
Preferably, the part of the connecting channel located in the sealing ring is in a shape of a "tesla valve" and the flow direction is towards the reaction tank.
Preferably, an anti-skid rubber pad is attached to the lower surface of the base plate.
Preferably, the anti-skid rubber pad is internally provided with a damping pore channel.
Preferably, the outer ring surface of the base disc is provided with an abrasion increasing channel.
The kit for identifying the human mesenchymal stem cells has the advantages that: in the scheme, the kit is used for identifying the differentiation direction of the human bone marrow mesenchymal stem cells, so that the phenomenon that the residual liquid, pancreatin solution and bone marrow mesenchymal stem cell inducing liquid are remained through external facilities such as a dropper in the prior art and the phenomenon that the liquid is easily polluted in the liquid material adding process is avoided, the success rate of the identification process of the differentiation direction of the human bone marrow mesenchymal stem cells in the bone marrow mesenchymal stem cells or the cartilage is greatly improved, the operation is simple and convenient, and the popularization and the application are facilitated.
Drawings
Fig. 1 is a sectional view of a kit for identifying human mesenchymal stem cells according to the present invention;
FIG. 2 is a top view of a basal disc of a kit for identifying human mesenchymal stem cells according to the present invention;
FIG. 3 is a top view of a sealing ring of a kit for identifying human mesenchymal stem cells according to the present invention;
fig. 4 is a sectional view of an anti-slip rubber pad of the kit for identifying human mesenchymal stem cells according to the present invention.
In the figure: the device comprises a base plate 1, a sealing ring 2, a threaded connecting ring 3, a mounting groove 4, a reaction tank 5, a reactant placing soft bag 6, an extrusion iron ball 7, an internal thread ring 8, an extrusion magnet rod 9, a connecting channel 10, a sealing cover 11, a clamping ring 12, an anti-skid rubber pad 13, a damping pore channel 14 and a grinding increasing channel 15.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", and the like, are used in the orientations and positional relationships indicated in the drawings, which are based on the orientations and positional relationships indicated in the drawings, and are used for convenience of description and simplicity of description, but do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention.
Example 1:
referring to fig. 1-4, a kit for identifying human mesenchymal stem cells, comprising a base plate 1, wherein a reaction tank 5 is arranged in the middle of the upper surface of the base plate 1, three mounting grooves 4 are arranged on the upper surface of the base plate 1 and on the outer side of the reaction tank 5 at intervals, a reactant placing soft bag 6 with a U-shaped cross section is filled in a part of the upper surface of the base plate 1 at one end of each mounting groove 4, the upper end of the reactant placing soft bag 6 is open and is flush with the upper surface of the base plate 1, the other end of each mounting groove 4 extends to the outer annular surface of the base plate 1, an internal thread ring 8 is embedded in a part of the mounting groove 4 close to the outer annular surface of the base plate 1, an extrusion magnet rod 9 is arranged on the internal thread ring 8 in a penetrating manner, one end of the extrusion magnet rod 9 extends to the outer annular surface of the base plate 1, and an extrusion iron ball 7 is arranged between one end of the extrusion magnet rod 9 in the base plate 1 and the reactant placing soft bag 6.
The middle part of the upper surface of the base plate 1 is provided with a sealing ring 2, the inner ring surface of the sealing ring 2 extends to the inner side of the ring groove surface of the reaction groove 5, and the diameter length of the inner ring of the sealing ring 2 is three quarters of the diameter of the upper end of the reaction groove 5; the lower surface of sealing ring 2 covers the upper end of mounting groove 4 and is close to the outside of 1 upper surface of basal disc, and the outside of 1 upper surface of basal disc corresponds the outer anchor ring of sealing ring 2 and is equipped with threaded connection ring 3, and threaded connection between the inner ring face of threaded connection ring 3 and the outer anchor ring face of sealing ring 2, and closely laminate between the upper surface of basal disc 1 and the lower surface of sealing ring 2.
The lower surface of the sealing ring 2 corresponding to the upper end of the reactant placing soft bag 6 is provided with a connecting channel 10, and the other end of the connecting channel 10 extends to the lower surface of the sealing ring 2 along the ring body of the sealing ring 2 and faces the reaction tank 5.
The upper surface of sealing ring 2 is equipped with joint ring 12, and sealing ring 2 has sealed lid 11 through joint ring 12 card lid, and the lower surface of sealed lid 11 closely laminates with the upper surface of sealing ring 2, and joint ring 12 makes and can closely laminate between sealed lid 11 and the sealing ring 2 to can realize the fixed to sealing ring 2 middle part through-hole.
The part of the connecting channel 10 positioned in the sealing ring 2 is in a 'Tesla valve' shape, the flowing direction faces the reaction tank 5, and in the process of putting the whole kit into a centrifuge for centrifugation, because the part of the connecting channel 10 positioned in the sealing ring 2 is in the 'Tesla valve' shape, and the flowing direction faces the reaction tank 5, the liquid in the reaction tank 5 can be prevented from flowing back to the reactant placing soft bag 6 through the connecting channel 10, thereby ensuring the smooth proceeding of centrifugation;
the anti-skid rubber pad 13 is attached to the lower surface of the base plate 1, the shock absorption hole 14 is formed in the anti-skid rubber pad 13, the anti-skid rubber pad 13 enables the reagent kit to be more stable in the placing process, and when the reagent kit is under downward pressure, the shock absorption hole 14 can play a shock absorption and buffering role, so that the phenomenon that the base plate 1 is damaged due to stress is avoided; increase and grind channel 15 has been seted up to the outer anchor ring of base plate 1, increases and grind channel 15 makes the kit by the base plate 1 carry out the centre gripping, hold the in-process of holding, better with the clamping part or hold the portion and keep stable.
Three reactant placing soft bags 6 are respectively provided with: removing culture solution from the 3 rd generation logarithmic growth phase bone marrow mesenchymal stem cells, washing with PBS for 2 times, collecting the residual solution, pancreatin solution with concentration of 0.25%, and bone marrow mesenchymal stem cell osteogenesis inducing solution.
In the application process, the differentiation process towards the bone direction is combined, the reactant placing soft bag 6 containing the residual liquid is extruded by rotating and extruding the magnet rod 9 according to the requirement of the identification direction of the human mesenchymal stem cells, and the residual liquid in the reactant placing soft bag 6 flows into the reaction tank 5 through the connecting channel 10;
further, a reaction agent containing pancreatin solution is placed in the soft bag 6 to be extruded through the rotary extrusion magnet rod 9, and at the moment, the pancreatin solution flows into the reaction tank 5 through the connecting channel 10, so that the digestion of the residual liquid in the reaction tank 5 is realized, and after the digestion is carried out for 10 minutes;
the DMEM medium containing 10% FBS required for terminating digestion in the identification process can be added into the reaction tank 5 by opening the sealing cover 11 to terminate digestion;
further, the sealing cover 11 is covered, the whole reagent kit is placed in a centrifuge for centrifugation at 1500r/min for 10 minutes, at this time, because the part of the connecting channel 10, which is positioned in the sealing ring, is in a shape of a 'Tesla valve' and the flowing direction faces to the reaction tank 5, the liquid in the reaction tank 5 can be prevented from flowing back to the reaction agent placing soft bag 6 through the connecting channel 10, and therefore the smooth operation of the centrifugation is ensured;
after the centrifugation is finished, the reaction agent containing the bone marrow mesenchymal stem cell osteogenesis inducing liquid is placed in the soft bag 6 to be extruded through the rotary extrusion magnet rod 9, at the moment, the operation of inducing the L-DMEM culture medium can be realized, namely, the bone marrow mesenchymal stem cell osteogenesis inducing liquid flows into the reaction tank 5 through the connecting channel 10, so far, in the process of identifying through the kit, the liquid material addition is realized through the rotary extrusion magnet rod 9, further, the traditional addition of the residual liquid, the pancreatic enzyme solution and the bone marrow mesenchymal stem cell osteogenesis inducing liquid through external facilities such as a dropper and the like is avoided, namely, the phenomenon of pollution easily caused in the liquid material adding process is avoided, the success rate of the identification process aiming at the bone formation direction of the human bone marrow mesenchymal stem cells is greatly improved, the operation is simple and convenient, and the popularization and application are facilitated.
Example 2
Referring to fig. 1 to 4, a kit for identifying human mesenchymal stem cells, which is different from example 1 in that three reaction agent-containing flexible bags 6 are respectively filled with: removing culture solution from the 3 rd generation logarithmic growth phase bone marrow mesenchymal stem cells, washing the culture solution with PBS for 2 times, and then obtaining residual solution, pancreatin solution with the concentration of 0.25 percent and bone marrow mesenchymal stem cells cartilage inducing solution.
In the application process, the differentiation process towards the cartilage direction is combined, the reactant placing soft bag 6 containing the residual liquid is extruded by rotating and extruding the magnet rod 9 according to the requirement of the identification direction of the human mesenchymal stem cells, and the residual liquid in the reactant placing soft bag 6 flows into the reaction tank 5 through the connecting channel 10;
further, a reaction agent containing pancreatin solution is placed in the soft bag 6 to be extruded through the rotary extrusion magnet rod 9, and at the moment, the pancreatin solution flows into the reaction tank 5 through the connecting channel 10, so that the digestion of the residual liquid in the reaction tank 5 is realized, and after the digestion is carried out for 10 minutes;
the DMEM medium containing 10% FBS required for terminating digestion in the identification process can be added into the reaction tank 5 by opening the sealing cover 11 to terminate digestion;
further, the sealing cover 11 is covered, the whole kit is placed in a centrifuge for centrifugation at 1500r/min for 5 minutes, at the moment, the part of the connecting channel 10, which is positioned in the sealing ring, is in a 'Tesla valve' shape, and the flowing direction faces to the reaction tank 5, so that the liquid in the reaction tank 5 can be prevented from flowing back to the reaction agent placing soft bag 6 through the connecting channel 10, and the smooth proceeding of the centrifugation is ensured;
after the centrifugation is finished, the soft bag 6 containing the reaction agent containing the mesenchymal stem cell chondrogenic induction liquid is placed and extruded through the rotary extrusion magnet rod 9, at the moment, the operation of inducing the L-DMEM culture medium can be realized, namely, the mesenchymal stem cell chondrogenic induction liquid flows into the reaction tank 5 through the connecting channel 10, so far, in the process of identifying through the kit, the liquid material addition is realized through the rotary extrusion magnet rod 9, further, the addition of the residual liquid, the pancreatic enzyme solution and the mesenchymal stem cell chondrogenic induction liquid through external facilities such as a dropper and the like is avoided, the phenomenon of pollution caused easily in the liquid material addition process is avoided, the success rate of the identification process aiming at the osteogenic direction of the mesenchymal stem cells of the human body is greatly improved, the operation is simple and convenient, and the popularization and the application are facilitated.
With the combination of the embodiment 1 and the embodiment 2, the method avoids the phenomenon that the residual liquid, the pancreatin solution and the mesenchymal stem cell inducing liquid are conventionally carried out through external facilities such as a dropper and the like in the process of identifying the osteogenic or chondrogenic differentiation direction of the mesenchymal stem cells of human, and the liquid material is easily polluted in the adding process, greatly improves the success rate of the identification process of the osteogenic or chondrogenic differentiation direction of the mesenchymal stem cells of human, is simple and convenient to operate, and is beneficial to popularization and application.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered as the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (8)
1. A kit for identifying human mesenchymal stem cells comprises a base plate (1) and is characterized in that a reaction groove (5) is formed in the middle of the upper surface of the base plate (1), three mounting grooves (4) are formed in the upper surface of the base plate (1) and in the outer side of the reaction groove (5) at intervals, a reactant placing soft bag (6) with a U-shaped section is filled in a part, located on the upper surface of the base plate (1), of one end of each mounting groove (4), the upper end of the reactant placing soft bag (6) is open and is flush with the upper surface of the base plate (1), the other end of each mounting groove (4) extends to the outer ring surface of the base plate (1), an internal thread ring (8) is embedded in a part, close to the outer ring surface of the base plate (1), and an extrusion magnet rod (9) penetrates through the internal thread ring (8), one end of the extrusion magnet rod (9) extends to the outside of the outer ring surface of the base plate (1), and an extrusion iron ball (7) is arranged between one end of the extrusion magnet rod (9) positioned in the base plate (1) and the reactant placing soft bag (6);
the middle part of the upper surface of the base plate (1) is provided with a sealing ring (2), the inner ring surface of the sealing ring (2) extends to the inner side of the ring groove surface where the reaction groove (5) is located, the lower surface of the sealing ring (2) covers the upper end of the mounting groove (4) and is close to the outer side of the upper surface of the base plate (1), the lower surface of the sealing ring (2) corresponding to the upper end of the reactant placing soft bag (6) is provided with a connecting channel (10), and the other end of the connecting channel (10) extends to the lower surface of the sealing ring (2) along the ring body of the sealing ring (2) and faces the reaction groove (5);
the three reactant placing soft bags (6) are respectively provided with: removing culture solution from the 3 rd generation logarithmic growth phase bone marrow mesenchymal stem cells, washing with PBS for 2 times, collecting the residual solution, pancreatin solution with concentration of 0.25%, and inducing solution of bone marrow mesenchymal stem cells.
2. The kit for identifying human mesenchymal stem cells according to claim 1, wherein the diameter length of the inner ring of the sealing ring (2) is three quarters of the diameter of the upper end of the reaction tank (5).
3. The kit for identifying human mesenchymal stem cells according to claim 1, wherein the upper surface of the sealing ring (2) is provided with a clamping ring (12), the sealing ring (2) is clamped and covered by a sealing cover (11) through the clamping ring (12), and the lower surface of the sealing cover (11) is tightly attached to the upper surface of the sealing ring (2).
4. The kit for identifying human mesenchymal stem cells according to claim 1, wherein a threaded connection ring (3) is arranged on the outer side of the upper surface of the base disc (1) corresponding to the outer annular surface of the sealing ring (2), an inner annular surface of the threaded connection ring (3) is in threaded connection with the outer annular surface of the sealing ring (2), and the upper surface of the base disc (1) is tightly attached to the lower surface of the sealing ring (2).
5. The kit for identifying human mesenchymal stem cells according to claim 1, wherein the portion of the connecting channel (10) located inside the sealing ring (2) is in a shape of a Tesla valve with the flowing direction facing the reaction tank (5).
6. The kit for identifying human mesenchymal stem cells according to claim 1, wherein a non-slip rubber pad (13) is attached to the lower surface of the basal disc (1).
7. The kit for identifying human mesenchymal stem cells according to claim 6, wherein the anti-skid rubber pad (13) is internally provided with a shock absorption hole (14).
8. The kit for identifying human mesenchymal stem cells according to claim 1, wherein the outer circumferential surface of the basal disc (1) is provided with an abrasion-increasing channel (15).
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| CN202111301926.0A CN114015552B (en) | 2021-11-04 | 2021-11-04 | Kit for identifying human mesenchymal stem cells |
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| CN105803534A (en) * | 2010-05-06 | 2016-07-27 | 艾比斯生物科学公司 | Integrated sample preparation system and stable enzyme mixture |
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| ES2752000T3 (en) * | 2005-05-09 | 2020-04-02 | Biofire Diagnostics Llc | Autonomous biological analysis |
| DE102007062441A1 (en) * | 2007-12-20 | 2009-06-25 | Aj Innuscreen Gmbh | Mobile rapid test system for nucleic acid analysis |
| JP6787924B2 (en) * | 2015-04-09 | 2020-11-18 | アングル ヨーロッパ リミテッド | Disposable bioassay cartridge, a method of performing multiple assay steps to transport fluid within the cartridge |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105803534A (en) * | 2010-05-06 | 2016-07-27 | 艾比斯生物科学公司 | Integrated sample preparation system and stable enzyme mixture |
| CN104593255A (en) * | 2015-02-06 | 2015-05-06 | 大连医科大学附属第二医院 | Microfluidic chip for instantly detecting EGFR (epidermal growth factor receptor) mutation |
| CN107129930A (en) * | 2017-06-09 | 2017-09-05 | 北京百康芯生物科技有限公司 | A kind of fully integrated detection of nucleic acids micro-fluidic chip and its application method |
| CN207793229U (en) * | 2017-11-27 | 2018-08-31 | 深圳华炎微测医疗科技有限公司 | Molecule diagnoses micro-fluidic chip and molecule diagnoses micro-fluidic chip system |
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Effective date of registration: 20230410 Address after: 225300 East side of Tai Road at the entrance of China Pharmaceutical City, Taizhou Medical High tech Industrial Development Zone, Jiangsu Province, and west side of the first floor of Building G23, Building 0003, north side of Xinyang Road Patentee after: Jiangsu Lu's Biotechnology Co.,Ltd. Patentee after: Lu Jian Address before: 550001 No.28 Guiyi street, Yunyan District, Guiyang City, Guizhou Province Patentee before: THE AFFILIATED HOSPITAL OF GUIZHOU MEDICAL University |
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| CI03 | Correction of invention patent |
Correction item: Patentee|Address|Patentee Correct: Affiliated Hospital of Guizhou Medical University|550001 No.28 Guiyi street, Yunyan District, Guiyang City, Guizhou Province False: Jiangsu Lu's Biotechnology Co.,Ltd.|225300 East side of Tai Road at the entrance of China Pharmaceutical City, Taizhou Medical High tech Industrial Development Zone, Jiangsu Province, and west side of the first floor of Building G23, Building 0003, north side of Xinyang Road|Lu Jian Number: 16-02 Volume: 39 |
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| CI03 | Correction of invention patent |