Automatic fat decellularization system and decellularization method

Abstract

According to the automatic fat decellularization system and the decellularization method, a truss, an electric clamping jaw and a weighing sensor are combined through a control system, and a series of operations such as transferring, weighing, cover opening, cover closing and solution injection of a centrifugal bottle are achieved. Compared with the prior art, a conventional preparation process at least needs to be operated by two persons, a third person is needed for weighing and re-checking, and a large amount of labor and operation time can be saved. And meanwhile, the processing time of a plurality of centrifugal bottles is difficult to control by manual operation, so that errors among batches are caused, and the product quality is reduced. And the preparation process is modularized, so that the operation error is reduced, and the production quality of the fat decellularized material is improved. In addition, according to the decellularization method provided by the invention, the conveying of the solution is in a closed state, so that the risk of introducing microorganisms due to multiple times of liquid change exposure is avoided.

Description

Technical Field

[0001] The present invention relates to the field of tissue engineering, and in particular to an automated fat decellularization system and a decellularization method. Background Art

[0002] Adipose decellularized matrix extracted from fat is used for in vitro adipose tissue engineering and in vivo adipose tissue regeneration. By injecting adipose decellularized matrix into the area in need of repair, it can replenish tissue volume, improve skin texture, and promote adipose tissue regeneration and repair. Adipose decellularized matrix is ​​suitable for a variety of plastic surgery filling needs, including filling facial wrinkles, nasolabial folds, scars, and soft tissue volume loss caused by trauma and aging. As a natural tissue engineering scaffold material, adipose decellularized matrix can accept stem cells and cells in the host body, allowing cells to grow, differentiate, and rebuild tissue.

[0003] The goal of tissue decellularization preparation protocols is to effectively remove all cellular and nuclear material while minimizing adverse effects on the structural composition, bioactivity, and mechanical integrity of the extracellular matrix. Decellularization protocols typically include physical, chemical, and enzymatic methods, using physical treatments or ionic solutions to dissolve cell membranes, followed by enzymatic treatment to separate cellular components from the ECM, detergents to solubilize cytoplasmic and nuclear cell components, and finally, to remove cellular debris from the tissue. After decellularization, all residual chemicals must be removed to avoid adverse reactions of the host tissue to the chemicals.

[0004] Existing adipose tissue decellularization technology is mainly completed through oscillation cleaning in a reaction vessel. Its batch processing volume is small, and the cleaning fluid is frequently replaced throughout the decellularization process, which easily increases the risk of contamination. Production quality control is difficult, which limits the large-scale production and application of adipose decellularized matrix.

[0005] The present invention provides a fully automated fat decellularization system and preparation method, which reduces manual intervention in the decellularization process of adipose tissue, reduces the risk of contamination, makes the quality of fat decellularized matrix more stable, increases batch processing volume, shortens processing time, and is conducive to industrial production. Summary of the Invention

[0006] The present application provides a fully automated fat decellularization system and preparation method, which reduces manual intervention in the decellularization process of adipose tissue, reduces the risk of contamination, makes the quality of fat decellularized matrix more stable, increases batch processing volume, shortens processing time, and is conducive to the realization of industrial production.

[0007] The present application discloses an automated fat decellularization system, characterized in that the system comprises: a sample loading system, a liquid adding system, a detection system, a mixing system, a centrifugal system and a control system, wherein:

[0008] The centrifugal system includes a centrifuge bottle temporary storage area and a constant temperature centrifuge, the centrifuge bottle temporary storage area is configured with a self-cleaning function, and the constant temperature centrifuge is configured inside the equipment;

[0009] The mixing system includes a constant temperature oscillation module, which is also configured inside the device and has an adjustable oscillation frequency;

[0010] The detection system includes a pH detection solution and a cleaning module, wherein the cleaning module is equipped with a plurality of pH probe cleaning tubes;

[0011] The electronic control system includes a display screen and a barcode scanner;

[0012] The loading system includes a three-axis truss, a two-axis truss, a pH probe, and an electric gripper; the three-axis truss is connected to the electric gripper and is configured to move in the X, Y, and Z directions within the installation range; the electric gripper is configured to complete the actions of clamping the bottle, flipping, and opening the cap; the two-axis truss is equipped with a filling needle;

[0013] The liquid adding system includes a solution bottle temporary storage area, a peristaltic pump, a weighing module and a filling needle bracket. The peristaltic pump is configured outside the equipment, and the silicone tube at the rear of the peristaltic pump is configured with a filter; the weighing module is configured with a weighing sensor;

[0014] The control system is used to set programs to start, stop and control the movement of components of other systems.

[0015] Optionally, in a preferred example, the system further includes an electronic control system, and the electronic control system includes a display screen and a barcode scanner.

[0016] Optionally, in a preferred example, the liquid adding system is further equipped with a filling needle holder.

[0017] Optionally, in a preferred example, the cleaning module is configured to perform pH detection using the pH calibration solution before each operation of the pH probe.

[0018] Optionally, in a preferred example, the constant temperature oscillation module is configured with an oscillation mold, and the oscillation mold includes an upright design and a horizontal design.

[0019] Optionally, in a preferred example, the peristaltic pump and the silicone tube are detachably assembled and the silicone tube is combed by a silicone tube slot during the connection process.

[0020] Optionally, in a preferred example, the filling needles and the solution temporary storage areas are numbered according to a corresponding relationship.

[0021] Optionally, in a preferred embodiment, the filter has a specification of 0.3-0.6 μm.

[0022] The present application also discloses an automated adipose tissue decellularization method, characterized in that the method comprises the following steps:

[0023] (1) Cleaning

[0024] Washing the adipose tissue to preliminarily remove impurities and blood, thereby obtaining washed adipose tissue, and placing the adipose tissue in a centrifuge bottle;

[0025] (2) Pretreatment

[0026] The rinsed fat tissue in the centrifuge bottle is centrifuged and homogenized by the constant temperature centrifuge in the centrifugal system, the fat cells are broken, the lipids are removed and the blood is further removed to obtain a solid layer; and then the centrifuge bottle is scanned by the applicable barcode scanner and placed in the centrifuge bottle temporary storage area, and the solution is injected into the container by the liquid adding system and placed in the mixing system by the electric clamp for mixing and shaking, and then the centrifuge bottle is placed in the centrifugal system by the electric clamp and centrifuged by the constant temperature centrifuge to obtain the rinsed solid layer.

[0027] (3) Virus inactivation

[0028] The centrifuge bottle in step (2) is placed into the mixing system through the electric gripper of the sample loading system, and the virus inactivation reagent is added through the liquid loading system for mixing and shaking. Then, the centrifuge bottle is placed into the centrifugal system through the electric gripper and centrifuged in the constant temperature centrifuge to obtain an inactivated solid layer;

[0029] wherein the virus inactivation treatment comprises mixing the solid layer with a virus inactivation reagent, wherein the virus inactivation reagent comprises an aqueous solution of a peroxide and a second alcohol;

[0030] (4) Decellularization

[0031] The solid layer in step S3 is treated by adding a decellularization reagent to the centrifuge bottle through the solution adding module and mixed and shaken. Then, the centrifuge bottle is placed into the centrifugal system through the electric clamp and centrifuged in the constant temperature centrifuge to finally obtain the target fat decellularized matrix.

[0032] Optionally, in a preferred embodiment, the adipose decellularized matrix can be pulverized to form particles.

[0033] Optionally, in a preferred embodiment, the adipose decellularized matrix can be added with injection water, physiological saline and PBS solution in different proportions to prepare adipose decellularized matrix suspension.

[0034] Alternatively, in a preferred embodiment, the adipose decellularized matrix can be prepared into emulsions of different concentrations, which are then injection molded and freeze-dried to obtain a dry adipose decellularized matrix.

[0035] Optionally, in a preferred example, the reagents used in the pretreatment, virus inactivation, and decellularization processes and other cleaning fluids such as purified water, sodium chloride solution, phosphate buffer, etc. are stored in a storage tank; the storage tank is connected to the peristaltic pump and the filling needle through a pipeline, and a filtering device is provided at the outlet of the storage tank to filter impurities and microorganisms in the solution.

[0036] Optionally, in a preferred embodiment, after each oscillation treatment, the system controls the pH probe to measure the pH of the solution after the oscillation treatment, and proceeds to the next step after reaching a preset range.

[0037] The advantages of this application are:

[0038] 1) The truss, electric gripper and weighing sensor are combined through a control system to realize a series of operations such as transfer, weighing, opening and closing the lid of the centrifuge bottle, and injecting the solution. The conventional preparation process requires at least two people to operate, and a third person is required to weigh and double-check. The present invention can save a lot of manpower and operation time. At the same time, manual operation makes it difficult to control the processing time of multiple centrifuge bottles, which will cause errors between batches and reduce the quality of the product. The preparation process is streamlined, which reduces operational errors and improves the production quality of fat decellularized materials.

[0039] 2) Since adipose tissue is a nutrient, microorganisms introduced during the treatment process are extremely easy to reproduce, and the solution delivery in the method applicable to this application is in a closed state, and 4. This system integrates cleaning, oscillation, centrifugation, solution addition and control modules in one, and all modules are connected to the clean room purification system through the system interface, ensuring that the entire decellularization process is in a purified state and avoiding the risk of multiple liquid changes exposing the introduction of microorganisms.

[0040] 3) This system has a pre-set automated decellularization program, and no manual operation is required during the process, which reduces the pollution caused by manual operation and avoids the mistakes caused by manual replacement of cleaning fluid. In addition, this application uses a barcode scanner to number the centrifuge bottles, realizing the full traceability of the fat decellularization preparation process.

[0041] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one.

[0042] The specification of this application records a large number of technical features, which are distributed in various technical solutions. If all possible combinations of technical features of this application (i.e., technical solutions) are to be listed, the specification will be too lengthy. In order to avoid this problem, the various technical features disclosed in the above-mentioned invention content of this application, the various technical features disclosed in the various embodiments and examples below, and the various technical features disclosed in the accompanying drawings can be freely combined with each other to form various new technical solutions (these technical solutions are all deemed to have been recorded in this specification), unless such a combination of technical features is technically infeasible. For example, in one example, feature A+B+C is disclosed, and in another example, feature A+B+D+E is disclosed. Features C and D are equivalent technical means that play the same role. Technically, only one of them can be used, and it is impossible to use them at the same time. Feature E can be technically combined with feature C. Then, the solution of A+B+C+D should not be considered as having been recorded because it is technically infeasible, while the solution of A+B+C+E should be considered as having been recorded. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 is a front view of an embodiment of the fully automatic fat decellularization system according to the present application;

[0044] Figure 2 is a top view of an embodiment of the fully automatic fat decellularization system according to the present application;

[0045] Figure 3 2 is a side view of an embodiment of the fully automatic fat decellularization system described in the present application.

[0046] Description of the accompanying drawings:

[0047] 1- Centrifuge bottle temporary storage area; 2- pH calibration solution; 3- Waste liquid port; 4- pH cleaning solution; 5- Centrifuge bottle gripper; 6- Constant temperature oscillation module; 7- Barcode scanner; 8- Sample loading system; 9- Constant temperature centrifuge; 10- Filling needle holder; 11- Peristaltic pump; 12- Solution bottle temporary storage area. DETAILED DESCRIPTION

[0048] After careful and in-depth research, the inventors have developed an automated fat decellularization system and decellularization method for the first time. Compared with the prior art, the present application combines a truss, an electric gripper and a weighing sensor through a control system to realize a series of operations such as transferring, weighing, opening and closing the centrifuge bottles, and injecting solutions. The conventional preparation process requires at least two people to operate, and a third person is required to weigh and double-check. The present invention can save a lot of manpower and operation time. At the same time, manual operation is difficult to control the processing time of multiple centrifuge bottles, which will cause errors between batches and reduce the quality of the product. The preparation process is streamlined, the operating errors are reduced, and the production quality of the fat decellularized material is improved. Not only that, the solution of the decellularization method proposed in the present application is transported in a closed state, which avoids the risk of multiple liquid changes exposing the introduction of microorganisms.

[0049] Example

[0050] In order to make the objectives, technical solutions and advantages of this application clearer, the implementation methods of this application will be further described in detail below with reference to the accompanying drawings.

[0051] The system in one embodiment described in this application is as follows Figure 1 As shown, specifically, the system includes: a workbench, a sample loading system, a liquid adding system, a detection system, a mixing system and a centrifugal system; wherein:

[0052] The centrifugal system includes a centrifuge bottle temporary storage area 1 and a constant temperature centrifuge 9, wherein the centrifuge bottle temporary storage area 1 is configured with a self-cleaning function, and the constant temperature centrifuge 9 is configured inside the device; optionally, in one embodiment, a limit position is configured under the temporary storage mold of the centrifuge bottle temporary storage area 1.

[0053] The mixing system includes a constant temperature oscillation module 6, which is also configured inside the device and has an adjustable oscillation frequency;

[0054] The detection system includes pH detection liquid and a cleaning module. The cleaning module is equipped with multiple pH probe cleaning tubes and pH cleaning liquid 4 and a waste liquid port 3. The cleaning module adopts a use-and-discharge mode of cleaning water to ensure the cleaning effect of the pH probe. The waste liquid port is used to dump the waste liquid of the pH probe cleaning liquid and the supernatant in the centrifuge bottle after centrifugation.

[0055] The electronic control system includes a display screen and a barcode scanner 7; the display screen can display the current device information; the electronic control system is configured with a setting program to start and stop and control the movement of other system components.

[0056] The loading system 8 includes a truss, a pH probe, an electric gripper and a centrifuge bottle gripper 5; in this embodiment, the truss includes a three-axis truss and a two-axis truss, the three-axis truss is made of corrosion-resistant material, is connected to the electric gripper, and is configured to move in the X, Y, and Z directions within the installation range; the electric gripper is made of 304 stainless steel and has a certain structural strength. The part in contact with the sample is equipped with a silicone pad for anti-slip and is configured to complete the actions of clamping the bottle, flipping, and opening the cover; the two-axis truss has a filling needle fixing slot to facilitate the fixing and removal of the filling needle, and can drive the filling needle to swing left and right and move up and down. The centrifuge bottle gripper 5 is used to fix the centrifuge bottle when the electric gripper is opening the cover

[0057] The liquid addition system includes a solution bottle temporary storage area 12, a peristaltic pump 11, a weighing module, and a filling needle holder 10. The filling needles of the two-axis truss are replaced by the filling needle holder 10. The peristaltic pump 11 is configured outside the equipment. The silicone tube at the rear of the peristaltic pump 11 is equipped with a filter. The weighing module is equipped with a weighing sensor, which is used to balance the centrifuge bottles and record the weight in the electronic batch record.

[0058] Optionally, in one embodiment, the cleaning module is configured to perform pH detection using the pH calibration solution before each operation of the pH probe.

[0059] Optionally, in one embodiment, the constant temperature oscillation module 6 is configured with an oscillation mold, and the oscillation mold includes an upright design and a horizontal design.

[0060] Optionally, in one embodiment, the peristaltic pump 11 and the silicone tube are detachably assembled and the silicone tube is combed by a silicone tube slot during the connection process.

[0061] Optionally, in one embodiment, the filling needles and the solution temporary storage areas are numbered in a corresponding relationship.

[0062] Optionally, in one embodiment, the filter has a specification of 0.3-0.6 μm.

[0063] This embodiment also discloses an automated adipose tissue decellularization method applicable to the automated adipose decellularization system, characterized in that the method comprises the following steps:

[0064] (1) Cleaning

[0065] Washing the adipose tissue to preliminarily remove impurities and blood, thereby obtaining washed adipose tissue, and placing the adipose tissue in a centrifuge bottle;

[0066] (2) Pretreatment

[0067] The rinsed fat tissue in the centrifuge bottle is centrifuged and homogenized by the constant temperature centrifuge 9 in the centrifugal system to break the fat cells, remove lipids and further remove blood to obtain a solid layer; and then the centrifuge bottle is scanned by the applicable barcode scanner 7 and placed in the centrifuge bottle temporary storage area 1, and the solution is injected into the container by the liquid adding system and placed in the mixing system by the electric clamp for mixing and shaking. Thereafter, the centrifuge bottle is placed in the centrifugal system by the electric clamp and centrifuged by the constant temperature centrifuge 9 to obtain the rinsed solid layer.

[0068] (3) Virus inactivation

[0069] The centrifuge bottle in step (2) is placed into the mixing system through the electric gripper of the sample loading system, and the virus inactivation reagent is added through the liquid loading system for mixing and shaking. Then, the centrifuge bottle is placed into the centrifugal system through the electric gripper and centrifuged in the constant temperature centrifuge 9 to obtain an inactivated solid layer;

[0070] wherein the virus inactivation treatment comprises mixing the solid layer with a virus inactivation reagent, wherein the virus inactivation reagent comprises an aqueous solution of a peroxide and a second alcohol;

[0071] (4) Decellularization

[0072] The solid layer in step S3 is treated by adding a decellularization reagent into the centrifuge bottle through the solution adding module and mixed and shaken. Then, the centrifuge bottle is placed into the centrifugal system through the electric clamp and centrifuged in the constant temperature centrifuge 9 to finally obtain the target fat decellularized matrix.

[0073] It should be noted that in the process of obtaining the solid layer by centrifugation in this embodiment, the system controls the three-axis truss and the electric gripper to transfer the centrifuge bottle to the centrifuge, and controls a certain temperature, time, and speed. After the centrifugation is completed, the system controls the three-axis truss and the electric gripper to transfer the centrifuge bottle to the waste liquid area, and the electric gripper completes the opening of the cover and the dumping of the waste liquid to obtain the solid layer at the bottom. The mixing and oscillation treatment is carried out by the system controlling the three-axis truss and the electric gripper to transfer the centrifuge bottle to a constant temperature oscillating shaker, controlling a certain temperature, time, and frequency, and performing the oscillation treatment. The centrifuge bottle temporary storage area 1 and the solution bottle temporary storage area 12 are both used for the turnover and temporary storage of corresponding components during the defatting process; the solution addition system in this method is completed by the two-axis truss and the filling needle thereon in conjunction with the three-axis truss and the electric gripper. During the injection process of pretreatment solutions, virus inactivation reagents, decellularization reagents, and other cleaning solutions such as purified water, sodium chloride solution, and phosphate buffer, the system controls a three-axis truss and electric gripper to transfer the centrifuge bottle to the weighing area, where the electric gripper opens the centrifuge bottle cap. A load cell measures the weight and calculates the required amount of solution to be added. A two-axis truss then moves the filling needle over the centrifuge bottle to inject the solution. Once injection is complete, the electric gripper closes the centrifuge bottle cap.

[0074] Optionally, in a preferred embodiment, the adipose decellularized matrix can be pulverized to form particles.

[0075] Alternatively, in one embodiment, the adipose decellularized matrix can be added to different proportions of water for injection, physiological saline, and PBS solution to prepare an adipose decellularized matrix suspension. Alternatively, the adipose decellularized matrix can be prepared into an emulsion of different concentrations, which can be injection molded and freeze-dried to obtain a dry adipose decellularized matrix.

[0076] Optionally, in one embodiment, the reagents used in the pretreatment, virus inactivation, and decellularization processes and other cleaning fluids such as purified water, sodium chloride solution, phosphate buffer, etc. are stored in a storage tank; the storage tank is connected to a peristaltic pump and a filling needle through a pipeline, and a filtering device is provided at the outlet of the storage tank to filter impurities and microorganisms in the solution.

[0077] Optionally, in one embodiment, after each oscillation treatment, the system controls the pH probe to measure the pH of the solution after the oscillation treatment, and proceeds to the next step after reaching a preset range.

[0078] Optionally, in one embodiment, the step of adding the solution can be arbitrarily performed before or after the transfer of the centrifuge bottle.

[0079] Optionally, in one embodiment, after performing the barcode scanning step of step (2) with the barcode scanning gun 7, the method responds to the obtained information through the control system to automatically execute the remaining steps.

[0080] It should be noted that in the application documents of this patent, relational terms such as first and second, etc. are merely used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "include," "comprise," or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article, or device comprising a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article, or device. Without further restriction, an element defined by the phrase "comprising a" does not exclude the presence of other identical elements in the process, method, article, or device comprising the element. In the application documents of this patent, if it is mentioned that an action is performed according to an element, it means that the action is performed at least according to the element, which includes two situations: performing the action only according to the element, and performing the action according to the element and other elements.

[0081] In this application, all directional indications (such as up, down, left, right, front, back, etc.) are only used to explain the relative position relationship, movement, etc. between the components under a certain specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indication will also change accordingly.

Claims

1. An automated fat decellularization system, characterized in that: The system includes: a sample loading system, a liquid adding system, a detection system, a mixing system, a centrifugal system and a control system, wherein: The centrifugal system includes a centrifuge bottle temporary storage area and a constant temperature centrifuge, wherein the centrifuge bottle temporary storage area is equipped with a self-cleaning function, and the constant temperature centrifuge is arranged inside the equipment; The mixing system includes a constant temperature oscillation module, which is also configured inside the device and has an adjustable oscillation frequency; The detection system includes a pH detection solution and a cleaning module, wherein the cleaning module is equipped with a plurality of pH probe cleaning tubes; The loading system includes a three-axis truss, a two-axis truss, a pH probe and an electric gripper; the three-axis truss is connected to the electric gripper and is configured to move in three directions of X, Y and Z within the installation range; the electric gripper is configured to complete the actions of clamping the bottle, flipping and opening the cover; the two-axis truss is equipped with a canning needle; The liquid adding system comprises a solution bottle temporary storage area, a peristaltic pump and a weighing module; the peristaltic pump is arranged outside the equipment, and the silicone tube at the rear of the peristaltic pump is arranged with a filter; the weighing module is arranged with a weighing sensor.

2. The automated fat decellularization system according to claim 1, characterized in that: The system also includes an electric control system, which includes a display screen and a barcode scanner and is configured with control software for setting programs to start, stop and move components of other systems.

3. The automated fat decellularization system according to claim 1, characterized in that: The liquid adding system is also equipped with a filling needle support.

4. The automated fat decellularization system according to claim 1, characterized in that: The cleaning module is configured to perform pH detection using the pH calibration solution before each operation of the pH probe.

5. The automated fat decellularization system according to claim 1, characterized in that: The constant temperature oscillation module is equipped with an oscillation mold, and the oscillation mold includes an upright design and a horizontal design.

6. The automated fat decellularization system according to claim 1, characterized in that: The peristaltic pump and the silicone tube are detachably assembled and the silicone tube is combed by the silicone tube slot during the connection process.

7. The automated fat decellularization system according to claim 1, wherein: The filling needles and the solution temporary storage areas are numbered according to a corresponding relationship.

8. The automated fat decellularization system according to claim 1, wherein: The specification of the filter is 0.3-0.6 μm.

9. An automated adipose tissue decellularization method applicable to the automated adipose decellularization system according to any one of claims 1 to 8, characterized in that: The method comprises the following steps: (1) Cleaning Washing the adipose tissue to preliminarily remove impurities and blood, thereby obtaining washed adipose tissue and placing the adipose tissue in a centrifuge bottle; (2) Pre-treatment The rinsed adipose tissue in the centrifuge bottle is centrifuged and homogenized by the constant temperature centrifuge in the centrifugal system to break the adipose cells, remove lipids and further remove blood to obtain a solid layer; and then the centrifuge bottle is scanned by the barcode scanner and placed in the centrifuge bottle temporary storage area, and the solution is injected into the container by the liquid adding system, and the electric clamp is used to put it into the mixing system for mixing and shaking, and centrifugation is performed to obtain the rinsed solid layer. (3) Virus inactivation The centrifuge bottle in step (2) is placed into the mixing system through the electric clamp of the sample loading system, and the virus inactivation reagent is added through the liquid loading system to perform mixing and shaking treatment, and then centrifuged to obtain the inactivated solid layer; Wherein, the virus inactivation treatment comprises mixing the solid layer with a virus inactivation reagent, wherein the virus inactivation reagent comprises an aqueous solution of a peroxide and a second alcohol substance; (4) Decellularization The solid layer in step S3 is treated by adding a decellularizing agent into a centrifuge bottle through a solution adding module, and then mixed and shaken, and then centrifuged to obtain the target fat decellularized matrix.

10. The automated adipose tissue decellularization method according to claim 9, characterized in that: The method is fully automated through a preset program.

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