CN113088433A

CN113088433A - Full-automatic cell processing pulsating workstation

Info

Publication number: CN113088433A
Application number: CN202110384182.7A
Authority: CN
Inventors: 李成龙; 邓慧婷; 王丹丹; 李南南; 吴迎梅; 骆莹; 朱争艳; 高英堂; 张自立; 张金卷
Original assignee: Tianjin Third Central Hospital
Current assignee: Tianjin Third Central Hospital
Priority date: 2021-04-09
Filing date: 2021-04-09
Publication date: 2021-07-09

Abstract

The invention relates to a full-automatic cell processing pulsating workstation and a control method, comprising the following steps: the system comprises a plurality of virus detection systems, a cell processing basic system, a plurality of rotary bin type incubators, a plurality of rotary bin type refrigerators, a plurality of rotary bin type heating boxes and a plurality of cell monitoring and culturing systems; the cell processing basic system comprises a biological safety cabinet and a workbench, wherein the workbench comprises an upper part and a lower part; the upper part of the workbench is provided with a connection unit, a quality control unit, a film sticking unit, a device with identification and clamping functions, a controller and a monitoring system; the under-counter portion includes a docking interface.

Description

Full-automatic cell processing pulsating workstation

Technical Field

The invention relates to the technical field of biomedicine, in particular to a full-automatic cell processing pulsating type workstation.

Background

In recent years, with the rapid development of cell biology, a large number of cell-based biomedical experiments and precise medical treatment projects, such as CAR-T, TCR-T, DC-CIK and the like, are emerged, and the integrity, activity, purity, density and the like of cells have more or less influence on the biomedical experiments. In addition, immune cell-based medical applications, particularly biotherapy, are rapidly developed in the field of precise medical treatment, so that the traditional medical treatment method is greatly influenced, and the unique personalized treatment and the target-specific precise treatment have the advantages of high safety, good specificity and definite curative effect, and more attention and application are paid. The main cell processing methods at present include both the magnetic bead method and the density gradient centrifugation method. The magnetic bead method may cause risks such as infection and embolism to human body due to the need of adding magnetic beads. The density gradient centrifugation treatment can be generally divided into plasma extraction inactivation, cell separation, cell washing and concentration, and cell culture, however, manual operation requires a long time, affects the efficiency of cell extraction, increases the risks of misoperation and pollution, and the activity of the extracted cells is affected due to an excessively long process. In addition, in the operation process, the hands of the sample and the personnel can enter and exit the biological safety cabinet for multiple times, so that the hidden dangers of sample pollution and environment pollution are increased. More importantly, the technology of different operators is different, and the consistency of quality cannot be guaranteed.

Disclosure of Invention

The invention aims at the problems that the manual density gradient centrifugation method has long cell processing time, low efficiency, mutual pollution of a sample and the environment and poor consistency, and provides a high-throughput standardized full-automatic cell processing pulsating workstation which replaces manual operation.

The invention discloses a full-automatic cell processing pulsating workstation, which comprises: the system comprises a plurality of virus detection systems, a cell processing basic system, a plurality of rotary bin type incubators, a plurality of rotary bin type refrigerators, a plurality of rotary bin type heating boxes and a plurality of cell monitoring and culturing systems; the cell processing basic system comprises a biological safety cabinet and a workbench, wherein the workbench comprises an upper part and a lower part; the upper part of the workbench is provided with a connection unit, a quality control unit, a film sticking unit, a device with identification and clamping functions, a controller and a monitoring system; the lower table portion comprises a docking interface;

the biological safety cabinet comprises an illuminating lamp and an ultraviolet lamp; the biological safety cabinet is provided with a plurality of special gas interfaces, a gas component control device, an air temperature control device, a corresponding infrared/wireless receiving device and an environment and operation state display screen, and can introduce carbon dioxide and nitrogen gas according to experiment needs and adjust the air temperature in the safety cabinet to meet different cell experiment needs and ensure cell activity; be equipped with the monitored control system that can emit infrared signal in the biological safety cabinet, can whole record experimentation, can carry out remote monitoring running state and adjustment biological safety cabinet internal environment or look back video recording so that trace to the source in the biological safety cabinet.

According to the full-automatic pulsating cell processing workstation, the quality control unit is a device for placing a virus microorganism detection plate, and a plurality of blood culture dishes can be placed for carrying out bacteria detection and HIV-HCV-TP-HBsAg quadruple cards for carrying out virus detection;

the connection unit is characterized in that a carrier driven by a motor can move in a reciprocating manner, and both sides of the biological safety cabinet are provided with switchable connection ports; the connecting unit can be in on-line object communication with equipment outside the single machine system, such as film pasting and transferring operations;

the film sticking unit has the advantages that the carrier driven by the motor can move in a reciprocating mode, the virus and microorganism detection plate on the carrier is placed into the film sticking machine to be subjected to film sticking treatment, and bidirectional pollution is prevented.

The full-automatic cell processing pulsating type workstation is characterized in that the rotating bin type incubator is arranged on a movable opening and closing panel arranged at the top inside the incubator, a first camera and a first fan are attached to the movable opening and closing panel, and a rotating bin position device is arranged inside a cabin body; the movable opening and closing panel is an interface for communicating the incubator with the outside; the first camera is used for identifying a sample; the first fan is used for quickly balancing the temperature of each position in the incubator and preventing the first camera from generating a water vapor guarantee identification function due to the fact that the temperature difference between the inside and the outside in a short time is caused by opening and closing the panel of the incubator; the rotating bin position device is convenient for grabbing samples and balancing the temperature in the incubator; the first camera is powered by the stable first power supply unit.

The full-automatic cell processing pulsating workstation is characterized in that the rotary bin type refrigerator is arranged on a movable opening and closing panel arranged at the top in the refrigerator, a second camera and a second fan are attached to the movable opening and closing panel, and a rotary bin device is arranged in a cabin body; the movable opening and closing panel is an external communication interface of the refrigerator; the second camera is used for identifying the sample; the second fan is used for quickly balancing the temperature of each position in the refrigerator, and is used for quickly cooling the sample to greatly reduce the solubility of denatured protein in the sample so as to ensure the infusion safety of the sample and prevent the second camera from generating a water vapor guarantee identification function due to the short-time internal and external temperature difference caused by opening and closing a refrigerator panel; the bin position rotating device is used for conveniently grabbing samples and balancing the temperature in the refrigerator; the second camera supplies power through the stable second power supply unit.

The full-automatic cell processing pulsating type workstation, first power supply unit and second power supply unit structure is the same, all includes: a switching tube M1-M36, resistors R1-R2 and R4, an adjustable resistor R3 and a capacitor C1; a first non-controllable end of the switching tube M1 is connected with a power supply VCC, a controllable end of the switching tube M1 is connected with a second non-controllable end of the switching tube M1 and a first non-controllable end of the switching tube M2, a controllable end of the switching tube M2 is connected with a second non-controllable end of the switching tube M2 and a first non-controllable end of the switching tube M3, a controllable end of the switching tube M3 is connected with a second non-controllable end of the switching tube M3, a controllable end of the switching tube M4, a first non-controllable end of the switching tube M4, a first non-controllable end of the switching tube M22, a first non-controllable end of the switching tube M24 and a first non-controllable end of the switching tube M26, and a second non-controllable end of the switching tube M4 is grounded;

the controllable end of the switch tube M5 and the first non-controllable end of the switch tube M8 are connected with a power supply VCC, the controllable end of the switch tube M5 is connected with the controllable end of the switch tube M8 and the second non-controllable end of the switch tube M5, the second non-controllable end of the switch tube M5 is connected with the first non-controllable end of the switch tube M6, the controllable end of the switch tube M6 is connected with the controllable end of the switch tube M9, the first non-controllable end of the switch tube M9 and the controllable end of the switch tube M12, the second non-controllable end of the switch tube M6 is connected with the first non-controllable end of the switch tube M7, the controllable end of the switch tube M7 is connected with the controllable end of the switch tube M10, the first non-controllable end of the switch tube M10, the controllable end of the switch tube M13 and the controllable end of the switch tube M17, the second non-controllable end of the switch tube M7 is connected with the second end of the resistor R4, and the second end of the resistor R4 is grounded; the controllable end of the switch tube M8 is connected with the controllable end of the switch tube M5, the second non-controllable end of the switch tube M8 is connected with the controllable end of the switch tube M12, the controllable end of the switch tube M16 and the first non-controllable end of the switch tube M9, the controllable end of the switch tube M9 is connected with the controllable end of the switch tube M6, the second non-controllable end of the switch tube M9 is connected with the controllable end of the switch tube M13, the controllable end of the switch tube M17 and the first non-controllable end of the switch tube M10, the controllable end of the switch tube M10 is connected with the controllable end of the switch tube M7, and the second non-controllable end of the switch tube M10 is grounded; the first non-controllable ends of the switch tube M11 and the switch tube M15 are connected with a power supply VCC, the controllable end of the switch tube M11 is connected with the second non-controllable end of the switch tube M15, the second non-controllable end of the switch tube M11 is connected with the first non-controllable end of the switch tube M12, the second non-controllable end of the switch tube M12 is connected with the first non-controllable end of the switch tube M13, the second non-controllable end of the switch tube M13 is connected with the first non-controllable end of the switch tube M14, the second non-controllable end of the switch tube M14 is grounded, the controllable end of the switch tube M14 is connected with the second non-controllable end of the switch tube M24, the first non-controllable end of the switch tube M25, the controllable end of the switch tube M26 and the controllable end of the switch tube M27, the controllable end of the switch tube M15 is connected with the second non-controllable end of the switch tube M11, the second non-controllable end 68628 is connected with the first non-controllable end of the switch tube M869 and the controllable end of the switch tube M8653, a second non-controllable end of the switching tube M16 is connected with a first non-controllable end of the switching tube M17, a controllable end of the switching tube M17 is connected with a first non-controllable end of the switching tube M10, a second non-controllable end of the switching tube M17 is connected with a first non-controllable end of the switching tube M18, a second non-controllable end of the switching tube M18 is grounded, and a controllable end of the switching tube M18 is connected with a second non-controllable end of the switching tube M26 and a first non-controllable end of the switching tube M27; the controllable end of the switching tube M22 is connected to the first output end of the control unit, the second non-controllable end of the switching tube M22 is connected to the controllable end of the switching tube M23, the first non-controllable end of the switching tube M23, the controllable end of the switching tube M24 and the controllable end of the switching tube M25, the second non-controllable end of the switching tube M23 is grounded, the controllable end of the switching tube M24 is connected to the controllable end of the switching tube M25 and the first non-controllable end of the switching tube M22, the second non-controllable end of the switching tube M24 is connected to the first non-controllable end of the switching tube M25 and the controllable end of the switching tube M14, the second non-controllable end of the switching tube M25 is grounded, the controllable end of the switching tube M26 is connected to the controllable end of the switching tube M27 and the first non-controllable end of the switching tube M24, and the second non-controllable end of the switching tube M26 is connected to the first controllable end of the switching tube M27 and the controllable end of the switching tube M27;

a first non-controllable end of the switch tube M19 is connected with a power supply VCC, a controllable end of the switch tube M19 is connected with a second non-controllable end of the switch tube M15, a second non-controllable end of the switch tube M19 is connected with a first non-controllable end of the switch tube M20, a controllable end of the switch tube M20 is connected with a first non-controllable end of the switch tube M20 and a first non-controllable end of the switch tube M21, a controllable end of the switch tube M21 is connected with a second non-controllable end of the switch tube M21, a second non-controllable end of the switch tube M21 is connected with a first non-controllable end of the switch tube M28, a first non-controllable end of the switch tube M29, a first non-controllable end of the switch tube M33 and a first non-controllable end of the switch tube M36, a controllable end of the switch tube M28 is connected with a controllable end of the switch tube M29, a second non-controllable end of the switch tube M69556, a second non-controllable end of the switch tube M8653, a second non-controllable resistor R8653 and a non-controllable end of the switch tube M8658, a controllable end of a switching tube M30 is connected with a second end of a resistor R2, a second non-controllable end of the switching tube M30 is connected with a first non-controllable end of a switching tube M31, a controllable end of a switching tube M31 and a controllable end of a switching tube M32, a second non-controllable end of the switching tube M31 is grounded, a controllable end of the switching tube M34 is connected with a second output end of the control unit, a second non-controllable end of the switching tube M34 is connected with a first end of a capacitor C1, a first non-controllable end of the switching tube M32 and a controllable end of a switching tube M35, a controllable end of the switching tube M32 is connected with a controllable end of a switching tube M31, a second non-controllable end of the switching tube M32 is grounded, a controllable end of the switching tube M33 is connected with a controllable end of a switching tube M29, a second non-controllable end of the switching tube M33 is connected with a controllable end of a switching tube M36, a second end of a capacitor C1, a second end of the controllable end of the switching tube M862 and a non-controllable end of the switching tube M36, the second end of the resistor R2 is connected with the first end of the adjustable resistor and the controllable end of the switch tube M30, and the second end of the adjustable resistor R3 is grounded.

The full-automatic cell processing pulsating type workstation is characterized in that the rotating bin type heating box is arranged on a movable opening and closing panel arranged at the top inside the heating box, a third camera and a third fan are attached to the movable opening and closing panel, and a rotating bin position device is arranged inside a cabin body; the movable opening and closing panel is an alternating current interface between the heating box and the outside; the third camera is used for identifying the sample; the third fan is used for quickly balancing the temperature of each position in the heating box and preventing the third camera from generating a water vapor guarantee identification function due to the short-time internal and external temperature difference caused by opening and closing the panel of the heating box; the rotating bin position device is used for conveniently grabbing samples and balancing the temperature in the heating box.

A method of controlling a fully automated cell processing pulsed workstation as claimed in any preceding claim, comprising: a stand-alone mode and an online mode.

The control method, the single machine mode includes: when a cell processing basic system is selected for independent use; manually placing a mixed sample of whole blood and anticoagulant in a sample unit, respectively placing related reagents, consumables and a gun head in the reagent unit, the consumables unit and the gun head unit, closing a front glass panel of the biological safety cabinet, and setting parameters such as temperature and humidity gas components required by an experiment; the two-dimensional codes on each pipe cap and each bottle cap are scanned and identified by each unit in the biological safety cabinet through the cap screwing device, whether the quantity and the type of articles of each unit meet the experimental requirements is checked according to a set experimental program, if the quantity and the type of the articles of each unit can not meet the experimental program, an alarm is given out, the operation is suspended for waiting for manual treatment, and if the quantity and the type of the articles of each unit meet the experimental program, the program is controlled to run by the controller;

if the experiment program is met, opening a window between the centrifuge and the workbench, moving the cover screwing device to the upper part of the sample unit for identification, and then grabbing the sample to the centrifuge, and if the experiment program is met, operating the rotor of the centrifuge to a specified position by the controller so that the cover screwing device can be aligned to place and grab the sample; if the sample self-balancing can not be met, the cover screwing device identifies, grabs the balancing pipe corresponding to the balancing unit and places the balancing pipe in the centrifuge for balancing;

after the placement, closing a window of the centrifuge, and performing centrifugal operation by the centrifuge;

after centrifugation, opening a window of the centrifuge, identifying and grabbing the sample to a sample unit by the cover screwing device, and grabbing to a balancing unit by the cover screwing device if a balancing pipe exists; the centrifuge window is closed. The sample unit clamps the sample tube, the reagent unit clamps the centrifuge tube, the consumable unit clamps the consumable tube, the screw cap device moves to the sample unit for identification and screw cap operation, then moves to the reagent unit for identification and screw cap operation, then moves to the consumable unit for identification and screw cap operation, then moves to the gun head unit for identification and screw cap (cap opening) and then resets, the liquid absorbing device moves to the gun head unit for assembling the gun head, then moves to the sample unit, the upper plasma of the sample is absorbed into the centrifuge tube of the consumable unit through the gun head, and then the residual plasma in the gun head moves to the quality control unit and drops into the virus detection card and the blood culture dish; then moving the gun head to the position above the waste recovery container device, discarding the gun head, then moving the gun head to the position of the gun head unit to assemble the gun head, then moving the reagent unit to absorb a reagent such as normal saline or PBS buffer solution, moving the reagent unit to the position of the sample unit, adding the sample into the sample, blowing, beating and mixing the sample uniformly, then absorbing and mixing the sample, adding the sample to the position of the reagent unit into a lymphocyte separation liquid centrifuge tube, then moving the reagent unit to the position above the waste recovery container device, discarding the gun head, and resetting the liquid; after the cover screwing device respectively moves to the sample unit, the reagent unit, the consumable unit and the gun head unit to perform cover screwing (cover closing) operation, the sample unit, the reagent unit and the consumable unit loosen the centrifugal tube; the cover screwing device identifies and grabs the plasma of the consumable unit to the heating unit, and the heating unit performs heating operation; opening a window of a centrifuge, identifying and grabbing a lymphocyte separation liquid tube in the reagent unit to the centrifuge by a screw cap device, clamping a corresponding balancing tube to the centrifuge if balancing is needed, closing the window of the centrifuge and performing centrifugation operation; the screw cap device moves into the sample unit to clamp the waste sample tube above the waste recovery container and reset after being discarded;

after centrifugation, a centrifugal machine window is opened, a lymphocyte separation liquid pipe is clamped to a reagent unit by a cover screwing device, the centrifugal machine window is closed, the reagent unit and a consumable unit clamp a centrifugal pipe, the cover screwing device respectively moves to the reagent unit, the consumable unit and a gun head unit to perform cover screwing (uncovering) operation and then resets, a liquid suction device moves to the gun head unit to assemble a gun head and then moves to the reagent unit, upper-layer liquid after lymphocyte separation is sucked into a new centrifugal pipe of the consumable unit through the gun head, then the gun head moves to the position above a waste recovery container device, and the gun head is discarded; then moving to a gun head unit to assemble a gun head, moving to a reagent unit to absorb a reagent such as normal saline or PBS buffer solution, moving to a consumable unit to add the reagent to the centrifugal tube, blowing, beating and mixing uniformly, moving the gun head to the position above a waste recovery container device, discarding the gun head, and resetting a liquid absorption device; the cover screwing device moves to the reagent unit, the consumable unit and the gun head unit respectively to perform cover screwing (cover closing) operation, and the reagent unit and the consumable unit loosen the centrifugal tube. Opening a window of the centrifuge, identifying and grabbing the centrifuge tube in the consumable unit to the centrifuge by the cover screwing device, clamping the corresponding balancing tube to the centrifuge if balancing is needed, closing the window of the centrifuge and performing centrifugal operation; the cover screwing device moves into the reagent unit to clamp the waste reagent tube above the waste recovery container and reset after being discarded;

the heating unit heats the plasma and then dissipates heat to reduce the solubility of denatured protein, reduce the possibility that cells are wrapped by fibronectin, improve the success rate and the amplification rate of cell culture, enhance the activity of cells, reduce the risk of adverse reactions such as thrombus and severe immune reaction in a reinfused human body, and enhance the curative effect of the cell infused human body on immune systems and tumor diseases;

opening a window of the centrifugal machine after centrifugation, identifying and taking out a centrifugal tube in the centrifugal machine to a sample unit by a cover screwing device, and identifying and taking out and placing in a balancing unit if a balancing tube exists; the sample unit and the reagent unit clamp the centrifuge tube, the screw cap device respectively moves to the sample unit, the reagent unit and the gun head unit to perform screw cap (cap opening) operation and then resets, the liquid absorbing device moves to the gun head unit to assemble the gun head, then the liquid absorbing device moves to the sample unit to absorb upper waste liquid, and the gun head is discarded. And then moving to a gun head unit to assemble the gun head, then moving to a reagent unit to absorb a reagent such as normal saline or PBS buffer solution, moving to a sample unit to add the reagent to the centrifugal tube, blowing and mixing the reagent, moving the gun head to the upper part of the waste recovery container device, discarding the gun head, and resetting the liquid absorption device. And after the cover screwing device respectively moves to the sample unit, the reagent unit and the gun head unit to perform cover screwing (cover closing) operation, the sample unit and the reagent unit loosen the centrifugal tube. The centrifugal machine window is opened, the cover screwing device identifies and grabs the centrifugal tube in the sample unit to the centrifugal machine, and the corresponding balancing tube is clamped to the centrifugal machine if balancing is needed, and the centrifugal machine window is closed and centrifugal operation is carried out. The screw cap device moves into the sample unit, clamps the waste reagent tube above the waste recovery container, and resets after discarding;

opening a window of the centrifugal machine after centrifugation, identifying and taking out a centrifugal tube in the centrifugal machine to a sample unit by a cover screwing device, and identifying and taking out and placing in a balancing unit if a balancing tube exists; the cover screwing device identifies and grabs a centrifugal tube in the heating unit and places the centrifugal tube in the centrifuge, if balancing is needed, the corresponding balancing tube is clamped to the centrifuge, a window of the centrifuge is closed, and freezing and centrifuging operations are carried out;

sample unit, reagent unit press from both sides the centrifuge tube tightly, and the spiral cover device moves reagent unit and consumptive material unit and rifle head unit respectively and carries out spiral cover (uncapping) operation back and resets, and the imbibition device moves and assembles the rifle head to rifle head unit department, then moves sample unit department, absorbs upper waste liquid through the rifle head, discards the rifle head. Then the liquid absorption device is moved to a gun head unit to assemble a gun head, then the liquid absorption device is moved to a reagent unit to absorb a reagent such as a cell culture medium, the reagent unit is moved to a sample unit to be added into the centrifugal tube to blow, beat and mix uniformly, the gun head is moved to the position above the waste recovery container device, the gun head is discarded, and then the liquid absorption device is reset. The cover screwing device moves to the reagent unit, the consumable unit and the gun head unit respectively to perform cover screwing (cover closing) operation, and then the reagent unit loosens the centrifugal tube.

The sample unit is subjected to oscillation operation;

and opening a window of the centrifuge after centrifugation, identifying and taking out a centrifuge tube in the centrifuge to a reagent unit by a cover screwing device, and identifying and taking out and placing in a balancing unit if a balancing tube exists. Sample unit, the reagent unit, the consumptive material unit is with the centrifuge tube, the blake bottle presss from both sides tightly, the spiral cover device removes the sample unit respectively, the reagent unit, the consumptive material unit, the rifle head unit resets after carrying out spiral cover (uncapping) operation, imbibition device removes rifle head to rifle head unit department assembly rifle head, then remove reagent unit department, get rid of denatured protein's plasma after drawing the centrifugation through the rifle head, add respectively in the new centrifuge tube of consumptive material unit and the centrifuge tube of sample unit and blow and beat the mixing, then shift the interior liquid transfer of centrifuge tube of sample unit to the blake bottle of suction consumptive material unit, and reserve a certain amount of liquid and shift to the quality control unit and instil into the blood culture dish and carry out the scribble board detection of reserving a kind. After the cover screwing device respectively moves to the sample unit, the reagent unit, the consumable unit and the gun head unit to perform cover screwing (cover closing) operation, the sample unit, the reagent unit and the consumable unit loosen the centrifuge tube and the culture bottle;

after the operation is finished, all the devices are reset.

The control method, the online mode includes: when a virus detection system, five cell processing basic systems, two rotating bin type incubators, one rotating bin type refrigerator, one rotating bin type heating box and one cell monitoring and culturing system are selected to form an online mode;

manually placing a mixed sample of whole blood and anticoagulant in a sample unit of a first cell processing basic system, respectively placing related reagents, consumables and gun heads in a reagent unit, a consumable unit and a gun head unit of each system, closing a front glass panel of the biological safety cabinet, and setting parameters such as temperature, humidity and gas components required by an experiment; the two-dimensional codes on each pipe cap and each bottle cap are scanned and identified by each unit in the biological safety cabinet through the cap screwing device, whether the quantity and the type of articles of each unit meet the experimental requirements is checked according to a set experimental program, if the quantity and the type of the articles of each unit can not meet the experimental program, an alarm is given out, the operation is suspended for waiting for manual treatment, and if the quantity and the type of the articles of each unit meet the experimental program, the program is controlled to run by the controller;

after centrifugation, the centrifuge window is opened, the sample is identified and grabbed to the sample unit by the cover screwing device, and if the balance pipe exists, the sample is grabbed to the balance unit by the cover screwing device. The centrifuge window is closed. The sample cell presss from both sides the sample cell tightly, the reagent unit presss from both sides the centrifuge tube tightly, the consumptive material unit presss from both sides tight back with the consumptive material pipe, the spiral cover device removes to move after the sample cell discerns and the spiral cover operation and identifies and move behind the spiral cover to the reagent unit and discern and spiral cover (uncap) the back and reset to rifle head unit, the imbibition device removes to rifle head unit department and assembles the rifle head, then remove sample unit department, absorb sample upper plasma to the centrifuge tube of consumptive material unit through the rifle head in, then remaining a certain amount of plasma removes to the unit of plugging into who is connected with virus detection system and instils into the virus detection card in the rifle head, in the blood culture dish. The connection device moves into the virus detection system, after the film is pasted by the film pasting unit, the connection device is grabbed to the quality control unit by a device with the functions of identification and clamping, then a new virus detection card and a blood culture dish in the quality control unit are clamped to the connection device, and the connection device moves to a first cell processing system for the next sample; the gun head is moved to the position above the waste recovery container device, the gun head is discarded, the gun head is moved to the position of the gun head unit to assemble the gun head, the reagent unit is moved to absorb a reagent such as normal saline or PBS buffer solution, the reagent unit is moved to the position of the sample unit to add the sample, the sample is blown, beaten and mixed uniformly, the mixed sample is absorbed to the position of the reagent unit and added into a lymphocyte separation liquid centrifuge tube, the sample unit is moved to the position above the waste recovery container device, the gun head is discarded, and the liquid absorbing device is. After the cover screwing device respectively moves to the sample unit, the reagent unit, the consumable unit and the gun head unit to perform cover screwing (cover closing) operation, the sample unit, the reagent unit and the consumable unit loosen the centrifugal tube; the cover screwing device identifies and grabs the plasma of the consumable unit to the rotary bin type heating box below the safety cabinet for heating operation. The connection window is opened, the cover screwing device identifies and grabs the lymphocyte separation liquid tube in the reagent unit to the connection unit, and the lymphocyte separation liquid tube is conveyed to the second cell processing basic system through the connection unit. The screw cap device moves into the sample unit to clamp the waste sample tube above the waste recovery container and reset after being discarded;

and the cover screwing device of the second cell processing basic system identifies and clamps the lymphocyte separation liquid tube on the connection unit into the centrifuge below the safety cabinet, and clamps the corresponding balancing tube to the centrifuge if balancing is needed, and the window of the centrifuge is closed and centrifugal operation is carried out. The screw cap device moves into the sample unit to clamp the waste sample tube above the waste recovery container and reset after the waste recovery container is discarded. Centrifuge window opens after the centrifugation, the spiral cover device presss from both sides lymphocyte separation liquid pipe and gets to the sample unit, the centrifuge window is closed, the sample unit, the reagent unit, the tight centrifuging tube of consumptive material unit clamp, the spiral cover device removes the sample unit respectively, reagent unit and consumptive material unit and rifle head unit reset after carrying out spiral cover (uncapping) operation, liquid suction device removes and assembles the rifle head to rifle head unit department, then remove reagent unit department, absorb upper liquid after the lymphocyte separation to the new centrifuging tube of consumptive material unit in through the rifle head, then the rifle head removes to waste recovery container device top, abandon the rifle head. Then the pipette tip is assembled at the pipette tip unit, then the pipette tip unit is moved to a reagent unit to suck a reagent such as normal saline or PBS buffer solution, the reagent unit is moved to a consumable unit to be added into the centrifugal tube just before blowing, beating and mixing, then the pipette tip is moved to the position above the waste recovery container device, the pipette tip is discarded, and then the liquid suction device is reset. The spiral cover device moves to sample unit, reagent unit, consumptive material unit, rifle head unit respectively and carries out spiral cover (close lid) operation back, and sample unit, reagent unit, consumptive material unit relax the centrifuge tube. And opening the connection window, identifying and grabbing the sample tube in the consumable unit to the connection unit by the cover screwing device, and conveying the sample tube to the third cell processing basic system through the connection unit. The screw cap device moves to the sample unit and the reagent unit, the waste sample tube is clamped above the waste recovery container and is reset after being discarded;

and the rotary cover device of the third cell processing basic system identifies and clamps the sample tubes on the connection unit into the centrifuge below the safety cabinet, and clamps the corresponding balancing tubes to the centrifuge if balancing is needed, and the window of the centrifuge is closed and centrifugal operation is carried out. After centrifugation, opening a window of the centrifuge, clamping the sample tube to the sample unit by the cover screwing device, and taking out the sample tube by the cover screwing device and placing the sample tube in the balancing unit if the balancing tube exists; centrifuge window is closed, and reagent unit, consumptive material unit press from both sides tight centrifuging tube, and the spiral cover device moves sample unit, reagent unit and consumptive material unit and rifle head unit respectively and carries out spiral cover (uncapping) operation back and resets, and the imbibition device moves to rifle head unit department and assembles the rifle head, then moves sample unit department, absorbs the upper waste liquid through the rifle head, discards the rifle head. And then moving to a gun head unit to assemble the gun head, then moving to a reagent unit to absorb a reagent such as normal saline or PBS buffer solution, moving to a sample unit to add the reagent to the centrifugal tube, blowing and mixing the reagent, moving the gun head to the upper part of the waste recovery container device, discarding the gun head, and resetting the liquid absorption device. After the cover screwing device respectively moves to the sample unit, the reagent unit, the consumable unit and the gun head unit to perform cover screwing (cover closing) operation, the sample unit, the reagent unit and the consumable unit loosen the centrifugal tube; and opening the connection window, identifying and grabbing the sample tubes in the sample units to the connection unit by the cover screwing device, and conveying the sample tubes to the fourth cell processing basic system through the connection unit. The cover screwing device moves into the reagent unit to clamp the waste reagent tube above the waste recovery container and reset after being discarded;

a screw cap device of the cell processing basic system II identifies and clamps the sample tubes on the connection unit into a centrifuge below the safety cabinet, clamps the corresponding balancing tubes to the centrifuge if balancing is needed, closes a window of the centrifuge and performs centrifugal operation; after centrifugation, opening a window of the centrifuge, clamping the sample tube to the sample unit by the cover screwing device, and taking out the sample tube by the cover screwing device and placing the sample tube in the balancing unit if the balancing tube exists; closing a centrifugal machine window, clamping a centrifugal tube by a reagent unit and a consumable unit, respectively moving a cover screwing device to a sample unit, the reagent unit, the consumable unit and a gun head unit to perform cover screwing (uncovering) operation and then reset, moving a liquid absorbing device to the gun head unit to assemble a gun head, then moving the liquid absorbing device to the sample unit, absorbing upper-layer waste liquid through the gun head, and discarding the gun head; and then moving to a gun head unit to assemble the gun head, then moving to a reagent unit to absorb a reagent such as normal saline or PBS buffer solution, moving to a sample unit to add the reagent to the centrifugal tube, blowing and mixing the reagent, moving the gun head to the upper part of the waste recovery container device, discarding the gun head, and resetting the liquid absorption device. After the cover screwing device respectively moves to the sample unit, the reagent unit, the consumable unit and the gun head unit to perform cover screwing (cover closing) operation, the sample unit, the reagent unit and the consumable unit loosen the centrifugal tube; and opening the connection window, identifying and grabbing the sample tubes in the sample units to the connection unit by the cover screwing device, and conveying the sample tubes to the fifth cell processing basic system through the connection unit. The cover screwing device moves into the reagent unit to clamp the waste reagent tube above the waste recovery container and reset after being discarded;

identifying and clamping the sample tubes on the connection unit into the sample unit by a rotary cover device of the cell processing basic system No. five, transferring plasma which is heated in a rotary bin type heating box below the cell processing system No. one into the cell processing system No. five through the connection unit of each cell processing basic system, identifying and clamping the plasma into a rotary bin type refrigerator below a safety cabinet by the rotary cover device for cooling, identifying and clamping the plasma into a centrifuge below the safety cabinet by the rotary cover device after cooling, clamping corresponding balancing tubes into the centrifuge if balancing is needed, closing a window of the centrifuge and carrying out refrigerated centrifugation; after centrifugation, opening a window of the centrifuge, clamping the sample tube to the sample unit by the cover screwing device, and taking out the sample tube by the cover screwing device and placing the sample tube in the balancing unit if the balancing tube exists; centrifuge window is closed, and sample unit, reagent unit, consumptive material unit press from both sides tight centrifuging tube and blake bottle, and the spiral cover device moves sample unit, reagent unit and consumptive material unit and rifle head unit respectively and carries out the spiral cover (uncapping) and operate the back and reset, and the imbibition device moves rifle head unit department and assembles the rifle head, then moves sample unit department, absorbs the upper waste liquid in the cell sample tube through the rifle head, discards the rifle head. And then moving to a gun head unit to assemble a gun head, moving to a sample unit, sucking the supernatant in the plasma sample tube to a new centrifuge tube of a consumable unit through the gun head, adding the residual certain amount into the cell sample tube, moving the gun head to the position above a waste recovery container device, and discarding the gun head. Then move to rifle head unit department and assemble the rifle head, then move to reagent unit and absorb reagent (culture medium) and add to sample unit and blow and beat in just cell sample tube evenly and absorb mixed liquid and move to the blake bottle of consumptive material unit, the rifle head moves to waste recovery container device top, abandons the rifle head, then imbibition device resets. After the cover screwing device respectively moves to the sample unit, the reagent unit, the consumable unit and the gun head unit to perform cover screwing (cover closing) operation, the sample unit, the reagent unit and the consumable unit loosen the centrifuge tube and the culture bottle; opening a connection window, identifying and grabbing culture bottles in the sample unit to the connection unit by the cover screwing device, and conveying the culture bottles into a cell monitoring culture system through the connection unit; the cover screwing device identifies and grabs the plasma sample in the consumable unit to a rotary bin type refrigerator below the safety cabinet for cold storage; the connecting unit screw cap device moves into the reagent unit to clamp the waste reagent tube above the waste recovery container and reset;

the cover screwing device of the cell monitoring culture system identifies and clamps the culture bottles on the connecting unit to the culture bottle carrier unit, the culture bottles are clamped by the carrier, the culture bottles are horizontally placed and loosened, and then the clamping device with the identification (used for clamping the culture bottles) identifies and clamps the culture bottles, shakes the culture bottles and counts the photos; then clamping the culture bottle into a rotary bin type incubator below the safety cabinet for culture;

after the operation is finished, all the devices are reset.

In order to solve the technical problems: the application provides and has solved standardization, uniformity, artifical problem, system architecture exquisiteness, the security is high, the function is various, the collocation is free, work efficiency is high, has greatly ensured the biosafety in the system and has shortened operating time and reduce intensity of labour, has reduced the technical demand to operating personnel, provides a more safe standardized solution for cell processing.

The invention has the advantages and positive effects that:

1. the invention relates to a more safe, reliable and convenient modularized cell processing system, which comprises a biosafety cabinet, a workbench, a balancing unit, a heating unit, a quality control unit, a connecting unit, a sample unit, a reagent unit, a consumable unit, a gun head unit, a waste unit, a liquid suction device, a controller, a screw cap device with identification and clamping functions, a controller and a centrifuge arranged below the workbench, this system structure is exquisite, the security is high, the function is various, the collocation is free, work efficiency is high, has got rid of most biological potential safety hazard, has greatly ensured the biological safety in the system and has shortened operating time and reduce intensity of labour, has reduced the technical demand to operating personnel, has got rid of the influence that operating personnel technical difference brought, provides a safer standardized solution for cell processing.

2. The invention completely contains the traditional flow of manually treating cells, all the steps can be manually replaced or replaced, so that the invention has extremely strong operation flexibility and flexible possibility, and particularly for special situations such as power failure, equipment failure and the like, the invention can be handed over by personnel to complete subsequent work; the system of the present invention can also be used to take over the subsequent work by manual operation. The operation process is changed from traditional open operation into closed operation, so that the possibility of pollution caused by various hidden dangers in the traditional manual mode is fundamentally eliminated, the safety and reliability of the whole system are further improved, the life safety of clinical patients using cells is more fully guaranteed, and the requirement that samples from different sources (people) are not uncapped at the same time is met; the screw cap design of all consumables and pipelines of the system ensures that the system is safe and reliable.

3. The biological safety cabinet comprises an illuminating lamp and an ultraviolet lamp. The window adopts the glass material to prevent that other materials like organic glass from producing the crackle and then taking place the potential safety hazard and influence the observation easily under ultraviolet and alcohol spray sterile operating mode, the glass window adopts electric drive lifting and falls, and the delivery window connecting device is reserved to the glass panels of both sides, can realize different functions and extension experiments by nimble various series connection equipment, can realize impulse type, pipelined operation. The biological safety cabinet is provided with a plurality of special gas interfaces, a gas component control device, an air temperature control device, a corresponding infrared/wireless receiving device and an environment and running state display screen, and can introduce gases such as carbon dioxide, nitrogen and the like according to experiment needs and adjust the air temperature in the safety cabinet so as to meet different cell experiment needs and guarantee cell activity. The monitoring system capable of emitting infrared signals is arranged in the cabinet, the experimental process can be recorded in the whole process, the running state of the system can be monitored remotely, and the environment in the biological safety cabinet can be adjusted or the video can be watched back so as to facilitate tracing.

4. The heating unit of the invention heats the plasma and then dissipates heat, so as to reduce the solubility of denatured protein, reduce the possibility that cells are wrapped by fibronectin, improve the success rate and the amplification rate of cell culture, enhance the activity of cells, reduce the risk of adverse reactions such as thrombus, severe immune reaction and the like in a reinfused human body, and enhance the curative effect of the cell infused human body on immune systems and tumor diseases.

5. The whole treatment process of the system is carried out in a closed manner in the system, and the blood and the sample of the patient are not in direct contact with the outside, so that the possibility that bacteria outside the system (such as a culture medium bottle body, operator gloves and the like) enter in the filling process is fundamentally avoided, and the pollution risk is greatly reduced; meanwhile, each consumable in the system is designed by adopting a threaded cover, so that the system is not opened, is safe and reliable, and realizes the possibility of opening and operating a single-source sample in the biological safety cabinet; meanwhile, the polluted air in the system cannot overflow to the outside of the system to pollute the environment.

6. An operator only needs to put sample reagent consumables in the system, the system can run in a full-automatic mode, manual operation is not needed in the whole process, labor intensity is greatly reduced, other work can be carried out by the operator, and labor time is greatly saved;

7. the liquid flow can be controlled by mechanical setting during sample adding, thereby eliminating individual difference of manual operation;

8. the system of the invention is closed operation in the system during cell treatment, thus fundamentally avoiding mutual pollution with the environment caused by open operation. The operator can monitor and adjust the operation parameters in real time through the remote control system without staying in front of the equipment, thereby greatly reducing the exposure risk to the operator, ensuring that the sample is not polluted by the operator in the processing process and ensuring that the inside and the outside of the system are safer.

9. The system can customize different combinations of the functional modules according to different requirements of users during implementation, and has strong practicability and wide application. Meanwhile, different medicines can be directly pre-installed in the system pipeline according to different requirements of a user during implementation, so that the step of allocating the medicines during operation of the user is omitted, the possibility of pollution is reduced, the system is safer, the labor intensity of the user is reduced, the labor time of the user is reduced, and the working efficiency is improved.

10. The invention realizes single machine, linkage combination and switching. The working mode realizes the switching of single-share operation, flow operation and pulse operation.

Drawings

FIG. 1 is a schematic diagram of a fully automated cell processing pulsed workstation according to the present invention.

FIG. 2 is a schematic diagram of the basic system of cell processing of the invention.

Fig. 3 is a schematic diagram of a power supply unit according to the present invention.

Detailed Description

The present application will now be described in further detail with reference to the drawings, it should be noted that the following detailed description is given for illustrative purposes only and is not to be construed as limiting the scope of the present application, as those skilled in the art will be able to make numerous insubstantial modifications and adaptations to the present application based on the above disclosure.

FIG. 1 is a schematic diagram of a fully automated cell processing pulsed workstation according to the present invention. The invention discloses a full-automatic cell processing pulsating workstation, which comprises: the system comprises a plurality of virus detection systems, a cell processing basic system, a plurality of rotary bin type incubators, a plurality of rotary bin type refrigerators, a plurality of rotary bin type heating boxes and a plurality of cell monitoring and culturing systems;

FIG. 2 is a schematic diagram of the basic system of cell processing of the invention. The cell processing basic system comprises a biological safety cabinet and a workbench; the workbench comprises an upper part 1 and a lower part 2; the upper part of the workbench comprises a plurality of unit function modules 4, and the unit function modules 4 are a balancing unit, a heating unit, a quality control unit, a connection unit, a sample unit, a reagent unit, a consumable unit, a gun head unit and a waste unit respectively; the liquid suction device 3, the first controller, the cover screwing device 5 with the functions of identification and clamping and the second controller, wherein the lower part of the platform comprises a centrifuge and a power supply control module, the biological safety cabinet is arranged outside the workbench, and a switch window is arranged on the biological safety cabinet;

Fig. 3 is a schematic diagram of the power supply unit according to the present invention. The full-automatic cell processing pulsating type workstation, first power supply unit and second power supply unit structure is the same, all includes: a switching tube M1-M36, resistors R1-R2 and R4, an adjustable resistor R3 and a capacitor C1; a first non-controllable end of the switching tube M1 is connected with a power supply VCC, a controllable end of the switching tube M1 is connected with a second non-controllable end of the switching tube M1 and a first non-controllable end of the switching tube M2, a controllable end of the switching tube M2 is connected with a second non-controllable end of the switching tube M2 and a first non-controllable end of the switching tube M3, a controllable end of the switching tube M3 is connected with a second non-controllable end of the switching tube M3, a controllable end of the switching tube M4, a first non-controllable end of the switching tube M4, a first non-controllable end of the switching tube M22, a first non-controllable end of the switching tube M24 and a first non-controllable end of the switching tube M26, and a second non-controllable end of the switching tube M4 is grounded;

The sample unit is subjected to oscillation operation;

after the operation is finished, all the devices are reset.

The invention has the advantages and positive effects that:

Claims

1. A fully automated cell processing pulsed workstation, comprising: the system comprises a plurality of virus detection systems, a cell processing basic system, a plurality of rotary bin type incubators, a plurality of rotary bin type refrigerators, a plurality of rotary bin type heating boxes and a plurality of cell monitoring and culturing systems; the cell processing basic system comprises a biological safety cabinet and a workbench, wherein the workbench comprises an upper part and a lower part; the upper part of the workbench is provided with a connection unit, a quality control unit, a film sticking unit, a device with identification and clamping functions, a controller and a monitoring system; the lower table portion comprises a docking interface;

2. The full-automatic cell processing pulsating workstation as claimed in claim 1, wherein the quality control unit is a device for placing a virus microorganism detection plate, and a plurality of blood culture dishes can be placed for bacterial detection and HIV-HCV-TP-HBsAg quadruple cards for virus detection;

3. The full-automatic cell processing pulsating workstation of claim 1, wherein the rotating chamber type incubator is provided with a movable opening and closing panel at the top of the inner part of the incubator, a first camera and a first fan are attached to the movable opening and closing panel, and a rotating chamber device is arranged in the chamber body; the movable opening and closing panel is an interface for communicating the incubator with the outside; the first camera is used for identifying a sample; the first fan is used for quickly balancing the temperature of each position in the incubator and preventing the first camera from generating a water vapor guarantee identification function due to the fact that the temperature difference between the inside and the outside in a short time is caused by opening and closing the panel of the incubator; the rotating bin position device is convenient for grabbing samples and balancing the temperature in the incubator; the first camera is powered by the stable first power supply unit.

4. The full-automatic pulsating cell processing workstation of claim 1, wherein said rotary bin refrigerator has a movable opening and closing panel on the top of the interior of the refrigerator, on which a second camera and a second fan are attached, and a rotary bin device is provided inside the chamber; the movable opening and closing panel is an external communication interface of the refrigerator; the second camera is used for identifying the sample; the second fan is used for quickly balancing the temperature of each position in the refrigerator, and is used for quickly cooling the sample to greatly reduce the solubility of denatured protein in the sample so as to ensure the infusion safety of the sample and prevent the second camera from generating a water vapor guarantee identification function due to the short-time internal and external temperature difference caused by opening and closing a refrigerator panel; the bin position rotating device is used for conveniently grabbing samples and balancing the temperature in the refrigerator; the second camera supplies power through the stable second power supply unit.

5. The fully automatic cell processing pulsating workstation of claim 3 or 4, wherein the first power supply unit and the second power supply unit are identical in structure and each comprise: a switching tube M1-M36, resistors R1-R2 and R4, an adjustable resistor R3 and a capacitor C1; a first non-controllable end of the switching tube M1 is connected with a power supply VCC, a controllable end of the switching tube M1 is connected with a second non-controllable end of the switching tube M1 and a first non-controllable end of the switching tube M2, a controllable end of the switching tube M2 is connected with a second non-controllable end of the switching tube M2 and a first non-controllable end of the switching tube M3, a controllable end of the switching tube M3 is connected with a second non-controllable end of the switching tube M3, a controllable end of the switching tube M4, a first non-controllable end of the switching tube M4, a first non-controllable end of the switching tube M22, a first non-controllable end of the switching tube M24 and a first non-controllable end of the switching tube M26, and a second non-controllable end of the switching tube M4 is grounded;

6. The full-automatic pulsating cell processing workstation of claim 1, wherein said rotating chamber type heating chamber has a movable opening and closing panel on the top of the interior of the heating chamber, on which a third camera and a third fan are attached, and the interior of the chamber body is provided with a rotating chamber position device; the movable opening and closing panel is an alternating current interface between the heating box and the outside; the third camera is used for identifying the sample; the third fan is used for quickly balancing the temperature of each position in the heating box and preventing the third camera from generating a water vapor guarantee identification function due to the short-time internal and external temperature difference caused by opening and closing the panel of the heating box; the rotating bin position device is used for conveniently grabbing samples and balancing the temperature in the heating box.

7. A method of controlling a fully automated cell processing pulsed workstation according to any of claims 1 to 6, comprising: a stand-alone mode and an online mode.

8. The control method of claim 7, wherein the stand-alone mode comprises: when a cell processing basic system is selected for independent use; manually placing a mixed sample of whole blood and anticoagulant in a sample unit, respectively placing related reagents, consumables and a gun head in the reagent unit, the consumables unit and the gun head unit, closing a front glass panel of the biological safety cabinet, and setting parameters such as temperature and humidity gas components required by an experiment; the two-dimensional codes on each pipe cap and each bottle cap are scanned and identified by each unit in the biological safety cabinet through the cap screwing device, whether the quantity and the type of articles of each unit meet the experimental requirements is checked according to a set experimental program, if the quantity and the type of the articles of each unit can not meet the experimental program, an alarm is given out, the operation is suspended for waiting for manual treatment, and if the quantity and the type of the articles of each unit meet the experimental program, the program is controlled to run by the controller;

sample unit, reagent unit press from both sides the centrifuge tube tightly, and the spiral cover device moves reagent unit and consumptive material unit and rifle head unit respectively and carries out spiral cover (uncapping) operation back and resets, and the imbibition device moves and assembles the rifle head to rifle head unit department, then moves sample unit department, absorbs upper waste liquid through the rifle head, discards the rifle head. Then the liquid absorption device is moved to a gun head unit to assemble a gun head, then the liquid absorption device is moved to a reagent unit to absorb a reagent such as a cell culture medium, the reagent unit is moved to a sample unit to be added into the centrifugal tube to blow, beat and mix uniformly, the gun head is moved to the position above the waste recovery container device, the gun head is discarded, and then the liquid absorption device is reset. The cover screwing device moves to the reagent unit, the consumable unit and the gun head unit respectively to perform cover screwing (cover closing) operation, and then the reagent unit loosens the centrifugal tube. The sample unit is subjected to oscillation operation;

after the operation is finished, all the devices are reset.

9. The control method of claim 7, wherein the online mode comprises: when a virus detection system, five cell processing basic systems, two rotating bin type incubators, one rotating bin type refrigerator, one rotating bin type heating box and one cell monitoring and culturing system are selected to form an online mode;

after the operation is finished, all the devices are reset.