阅读说明：本技术 细胞工厂生物反应器显微光电监测系统 (Micro-photoelectric monitoring system for bioreactor of cell factory ) 是由 宫平 张宁 马辰昊 朱海焕 葛辉琼 阚宝慧 郭红壮 吴昊 李旭 吉翔宇 谭国桢 于 2019-11-19 设计创作，主要内容包括：本发明公开了细胞工厂生物反应器显微光电监测系统,涉及生物反应器技术领域,包括计算机、细胞工厂生物反应器、倾斜显微光学系统、透射照明系统和X轴移动平台,细胞工厂生物反应器安装在X轴移动平台上以控制其在水平方向上左右移动,倾斜显微光学系统和透射照明系统分别安装在C型机械臂的两端,透射照明系统发出的光倾斜投射细胞工厂生物反应器后被倾斜显微光学系统接收到,并将生成的图像发送给计算机。本发明将显微光学系统安装在生物反应器的一侧,并采用倾斜透射的方式进行照明和图像采集,可以用来监测生物反应器各层培养叠板不同位置处细胞的生长状态,极大的扩大了监测范围,为相关科学研究提供了真实可靠的数据基础。(The invention discloses a microscopic photoelectric monitoring system of a cell factory bioreactor, which relates to the technical field of bioreactors and comprises a computer, a cell factory bioreactor, an inclined microscopic optical system, a transmission illumination system and an X-axis moving platform, wherein the cell factory bioreactor is arranged on the X-axis moving platform to control the cell factory bioreactor to move left and right in the horizontal direction, the inclined microscopic optical system and the transmission illumination system are respectively arranged at two ends of a C-shaped mechanical arm, and light emitted by the transmission illumination system is obliquely projected on the cell factory bioreactor and then received by the inclined microscopic optical system and sends the generated image to the computer. The invention installs the microscopic optical system at one side of the bioreactor, and adopts the inclined transmission mode to carry out illumination and image acquisition, thereby being capable of monitoring the growth state of cells at different positions of each layer of culture laminated plate of the bioreactor, greatly enlarging the monitoring range and providing a real and reliable data base for related scientific research.)

1. The cell factory bioreactor micro-photoelectric monitoring system is characterized by comprising a computer, a servo controller, a cell factory bioreactor, an inclined microscopic optical system, a cold light source illuminating system, an X-axis moving platform and an image acquisition system, wherein the cold light source illuminating system is used for controlling the working state of a transmission illuminating system;

the cell factory bioreactor is arranged on the X-axis moving platform, the inclined microscopic optical system and the transmission illumination system are respectively arranged at two ends of a C-shaped mechanical arm, the inclined microscopic optical system and the transmission illumination system are oppositely arranged, and the C-shaped mechanical arm is arranged on the Z-axis moving platform;

the X-axis moving platform is used for controlling the cell factory bioreactor to move in the horizontal direction, the Z-axis moving platform is used for controlling the C-shaped mechanical arm to move in the vertical direction, the C-shaped mechanical arm is in an inclined state, the inclined microscopic optical system and the transmission illumination system are respectively positioned on two sides of the cell factory bioreactor, and light emitted by the transmission illumination system is obliquely transmitted through the cell factory bioreactor and then received by the inclined microscopic optical system;

the image collected by the inclined microscopic optical system is transmitted to a computer through the image collecting system, the cold light source illuminating system controls the working state of the transmission illuminating system under the control of the computer, and the servo controller controls the moving states of the X-axis moving platform and the Z-axis moving platform under the control of the computer.

2. The cell factory bioreactor micro-electro-optical monitoring system of claim 1, further comprising a vertical micro-optical system, wherein the cold light source illumination system is further used for controlling the operation state of the reflection illumination system under the control of a computer;

the vertical microscopic optical system and the reflective lighting system are both arranged on a W-axis moving platform, the W-axis moving platform is arranged on a Y-axis moving platform, the Y-axis moving platform is used for controlling the W-axis moving platform to move in the horizontal direction, the W-axis moving platform is used for controlling the vertical microscopic optical system and the reflective lighting system to move in the vertical direction, the vertical microscopic optical system and the reflective lighting system are both positioned below the cell factory bioreactor and face the cell factory bioreactor, light emitted by the reflective lighting system irradiates the bottom of the cell factory bioreactor, and is received by the vertical microscopic optical system after being reflected;

the image collected by the vertical micro optical system is transmitted to a computer through the image collecting system, and the servo controller controls the moving states of the Y-axis moving platform and the W-axis moving platform under the control of the computer.

3. The cell factory bioreactor micro-electro-optical monitoring system of claim 2, wherein the X-axis moving platform, the Y-axis moving platform, the Z-axis moving platform and the W-axis moving platform each comprise a servo motor and a ball screw, the servo motors operate under the control of a servo controller, and the ball screws in the X-axis moving platform, the Y-axis moving platform, the Z-axis moving platform and the W-axis moving platform respectively drive the cell factory bioreactor, the W-axis moving platform, the C-shaped mechanical arm, the vertical microscopic optical system and the reflective illumination system to move under the driving of the servo motors.

4. The cell factory bioreactor micro-electro-optical monitoring system according to claim 3, wherein the servo motors are AC servo motors, each of the AC servo motors is provided with an encoder for monitoring the rotation angle of the AC servo motor and feeding back the rotation angle data obtained by monitoring to the computer.

5. The cell factory bioreactor micro-electro-optical monitoring system of claim 3, wherein the servo controller controls the servo motor through a control loop, the control loop comprises a position loop, a speed loop and a current loop, the position loop and the speed loop are arranged in parallel, the position command and the speed command are respectively subjected to PI processing and then are superposed with commands generated by speed feedforward control and acceleration feedforward control to form a current command, and the current command is subjected to PI processing and then is used for controlling the servo motor.

Technical Field

The invention relates to the technical field of bioreactors, in particular to a microscopic photoelectric monitoring system of a bioreactor in a cell factory.

Background

The bioreactor is a device for obtaining a desired product through biological reaction or self metabolism by realizing in vitro culture by simulating in vivo growth environment of enzymes or organisms (such as cells, microorganisms and the like). The bioreactor plays an important role in vaccine production, monoclonal antibody preparation, medicine production, tumor prevention and treatment, wine brewing, biological fermentation, organic pollutant degradation and the like.

Aiming at the limitation of the monitoring technology and the monitoring device for the growth state of the cultured cells in the bioreactor of the cell factory, the growth state of the cells at the bottommost layer is observed mainly by using an inverted microscope. The inverted microscope is mainly used for observing the surface morphology and the like of a monolayer culture dish, a glass slide or other objects, the microscope lens is positioned below the objective table, the illumination light source is positioned above the objective table, and the observed cell culture dish is arranged on the objective table. The monitoring method is limited by the optical performance of the traditional inverted microscope, has short working distance, can only observe the growth state of cells at the bottommost layer, can not take out adherent cells of other layers, can only be estimated by experience, can not be monitored visually, has great uncertainty in research and production, and can not ensure the quality.

Disclosure of Invention

The embodiment of the invention provides a microscopic photoelectric monitoring system of a bioreactor of a cell factory, which can solve the problems in the prior art.

The invention provides a micro-photoelectric monitoring system of a cell factory bioreactor, which comprises a computer, a servo controller, a cell factory bioreactor, an oblique microscopic optical system, a cold light source illuminating system, an X-axis moving platform and an image acquisition system, wherein the cold light source illuminating system is used for controlling the working state of a transmission illuminating system;

the cell factory bioreactor is arranged on the X-axis moving platform, the inclined microscopic optical system and the transmission illumination system are respectively arranged at two ends of a C-shaped mechanical arm, the inclined microscopic optical system and the transmission illumination system are oppositely arranged, and the C-shaped mechanical arm is arranged on the Z-axis moving platform;

the X-axis moving platform is used for controlling the cell factory bioreactor to move in the horizontal direction, the Z-axis moving platform is used for controlling the C-shaped mechanical arm to move in the vertical direction, the C-shaped mechanical arm is in an inclined state, the inclined microscopic optical system and the transmission illumination system are respectively positioned on two sides of the cell factory bioreactor, and light emitted by the transmission illumination system is obliquely transmitted through the cell factory bioreactor and then received by the inclined microscopic optical system;

the image collected by the inclined microscopic optical system is transmitted to a computer through the image collecting system, the cold light source illuminating system controls the working state of the transmission illuminating system under the control of the computer, and the servo controller controls the moving states of the X-axis moving platform and the Z-axis moving platform under the control of the computer.

The cell factory bioreactor microscopic photoelectric monitoring system comprises a computer, a cell factory bioreactor, an inclined microscopic optical system, a transmission illumination system and an X-axis moving platform, wherein the cell factory bioreactor is installed on the X-axis moving platform to control the cell factory bioreactor to move left and right in the horizontal direction, the inclined microscopic optical system and the transmission illumination system are respectively installed at two ends of a C-shaped mechanical arm, the C-shaped mechanical arm is installed on a Z-axis moving platform to control the cell factory bioreactor to move in the vertical direction, light emitted by the transmission illumination system is obliquely projected on the cell factory bioreactor and then received by the inclined microscopic optical system, and a generated image is sent to the computer. The invention installs the microscopic optical system at one side of the bioreactor, and adopts the inclined transmission mode to carry out illumination and image acquisition, thereby being capable of monitoring the growth state of cells at different positions of each layer of culture laminated plate of the bioreactor, greatly enlarging the monitoring range and providing a real and reliable data base for related scientific research.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a schematic diagram of the composition of a micro-photoelectric monitoring system of a bioreactor of a cell factory;

FIG. 2 is a schematic diagram of the structure of a micro-photoelectric monitoring system of a bioreactor of a cell factory;

FIG. 3 is a schematic diagram of a servo control system;

FIG. 4 is a schematic diagram of the overall structure of the controller;

FIG. 5 is a schematic diagram of a prior art PID cascade control loop;

FIG. 6 is a schematic diagram of a parallel speed and position PI control loop of the present invention;

fig. 7 is a detailed structural diagram of a speed and position parallel PI control loop.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1 and 2, the present invention provides a microscopic photoelectric monitoring system of a cell factory bioreactor, which comprises a computer 100, a servo controller 110, a cell factory bioreactor 122, a microscopic optical system, a cold light source illumination system, a feeding system and an image acquisition system, wherein the microscopic optical system comprises an inclined microscopic optical system 161 and a vertical microscopic optical system 151, the cold light source illumination system is used for controlling the working state of a reflection illumination system 152 and a transmission illumination system 162, the feeding system comprises an X-axis moving platform 120, the cell factory bioreactor 122 is mounted on the X-axis moving platform 120, the inclined microscopic optical system 161 and the transmission illumination system 162 are respectively mounted at two ends of a C-shaped mechanical arm 160, the inclined microscopic optical system 161 and the transmission illumination system 162 are oppositely arranged, the C-shaped mechanical arm 160 is mounted on the Z-axis moving platform 140, the vertical micro-optical system 151 and the reflective illumination system 152 are mounted on a W-axis moving stage 150, and the W-axis moving stage 150 is mounted on a Y-axis moving stage 130.

The X-axis moving platform 120 is configured to control the cell factory bioreactor 122 to move left and right in the horizontal direction, the Y-axis moving platform 130 is configured to control the W-axis moving platform 150 to move back and forth in the horizontal direction, the W-axis moving platform 150 is configured to control the vertical micro-optical system 151 and the reflective lighting system 152 to move in the vertical direction synchronously, the Z-axis moving platform 140 is configured to control the C-type robot 160 to move in the vertical direction, the C-type robot 160 is in an inclined state, the inclined micro-optical system 161 and the transmissive lighting system 162 are respectively located at two sides of the cell factory bioreactor 122, light emitted by the transmissive lighting system 162 is received by the inclined micro-optical system 161 after being transmitted through the cell factory bioreactor 122 in a downward inclined manner, so as to locally monitor edges of layers at the side of the cell factory bioreactor 122, at the same time, the C-arm 160 is rotated about a central axis 163 to rotate the tilt microscopy optical system 161 from one side of the cell factory bioreactor 122 to the other side to monitor the culture stack on the other side of the cell factory bioreactor 122. The vertical micro-optical system 151 and the reflected illumination system 152 are both located below the cell factory bioreactor 122 and the vertical micro-optical system 151 and the reflected illumination system 152 are both directed towards the cell factory bioreactor 122, and light emitted by the reflected illumination system 152 is irradiated at the bottom of the cell factory bioreactor 122, reflected and received by the vertical micro-optical system 151, so as to perform global monitoring on 1-3 layers at the bottom of the cell factory bioreactor 122. The computer 100 has a touch display screen to input various control commands. The images collected by the vertical microscope optical system 151 and the oblique microscope optical system 161 are transmitted to the computer 100 through the image collecting system, and the computer 100 also sends a control command to the image collecting system to control the image collecting process. The cold light source illumination system performs operation state control on the reflective illumination system 152 and the transmissive illumination system 162 under the control of the computer 100.

The X-axis motion stage 120, the Y-axis motion stage 130, the Z-axis motion stage 140, and the W-axis motion stage 150 each include a servo motor and a ball screw, the servo motors operate under the control of a servo controller, the ball screws drive the W-axis motion stage 150, the vertical micro-optical system 151, the reflective lighting system 152, the cell factory bioreactor 122, and the C-arm 160 to move under the drive of the servo motors, and the servo controller operates under the control of the computer 100.

The working process of the microscopic photoelectric monitoring system of the bioreactor of the cell factory is as follows: the X-axis moving platform 120 moves to the loading and unloading position of the cell factory bioreactor 122, the experimenter places the cell factory bioreactor 122 on the carrying tray 121, and the carrying tray 121 on the X-axis moving platform 120 moves to carry the cell factory bioreactor 122 to the monitoring position. And switching an inclined microscopic monitoring mode and a vertical microscopic monitoring mode, wherein in the inclined microscopic monitoring mode, a CCD camera on an inclined microscopic optical system 161 works, a transmission illumination system 162 corresponding to the CCD camera is turned on, the Z-axis moving platform 140 drives a C-shaped mechanical arm 160 to move in the vertical direction, and after the number of monitoring layers is selected, the X-axis moving platform 120 bears the cell factory bioreactor 122 to monitor the cell growth states of different positions of the layer of culture laminated plate. In the vertical microscopic monitoring mode, the CCD camera on the vertical microscopic optical system 151 works, the corresponding reflection illumination system 152 is turned on, the W-axis moving platform 150 controls the vertical microscopic optical system 151 to move in the vertical direction, the X-axis moving platform 120 moves the cell factory bioreactor 122 after the number of monitoring layers is selected, and the Y-axis moving platform 130 moves the vertical microscopic optical system 151, so that the overall monitoring of the growth state of cells in the bottom 1-3 layers of culture stacked plates is realized.

The mechanical structure of the cell factory bioreactor micro-photoelectric monitoring system belongs to three-dimensional 4-axis motion, and the C-shaped mechanical arm 160 needs to rapidly move to a specified monitoring layer and is accurately positioned, so that a power source adopts an alternating current servo motor which is small in inertia and suitable for high-speed and high-torque work, each servo motor is provided with a 17-bit encoder, the pulse equivalent is 9.888' to monitor the rotation angle of the servo motor, and the step-out problem of the traditional stepping motor can be solved. The ball screws of the moving platforms of each axis are controlled by the servo controllers to rotate corresponding servo motors, so that linear motion of the linear slide rails is realized, the rotating speed, the position and the like of the servo motors are controlled in real time, and the moving platforms of each axis are controlled by upper computer software to finish corresponding actions according to the sent motion instructions. The structure of a servo control system of the cell factory bioreactor micro-photoelectric monitoring system is shown in fig. 3, and an X-axis motor driver, a Y-axis motor driver, a Z-axis motor driver and a W-axis motor driver in fig. 3 are collectively called as a servo controller.

The inertia of the liquid medium during the movement of the cell factory bioreactor 122 affects the target control, and the vigorous shaking also affects the growth of the cells, so that the stability of the movement control is required to be high. In the conventional control, the PID control is one of the most common control methods, and the control error can be reduced by adjusting the proportional coefficient, the integral time constant and the differential time constant, so that the steady-state error can be reduced. However, when the control target changes greatly, a relatively large overshoot is generated in the control process, and in order to reduce the overshoot of the control, through analyzing the traditional PID control algorithm, the PI and feedforward control method is obtained through optimization and improvement, the overshoot under quick response is reduced, and the algorithm can ensure accurate positioning and realize good following performance.

The servo controller selects an alternating current servo controller with a main control chip being an ARM Cortex-M4 kernel STM32F407, and the overall structural block diagram of the controller is shown in FIG. 4 according to the characteristics of chip interfaces and control requirements.

The control of the servo motor is mainly realized by a position loop, a speed loop and a current loop, the control stability can be improved and the faster following performance can be kept by controlling the current loop and the speed loop, and the control positioning capability can be improved and the better position tracking performance can be kept by controlling the position loop.

A conventional cascade PID control structure is shown in fig. 5, in which a speed loop is nested in a position loop, and in the control structure, the position loop is usually controlled by proportional control, and the speed and current loops are controlled by proportional integral control, which is not favorable for fast adjustment. Therefore, the present invention uses a PI control and feedforward control mode with parallel speed and position, the control loop is as shown in fig. 6, the servo controller controls the servo motor through the control loop, the control loop includes a position loop, a speed loop and a current loop, the position loop and the speed loop are configured in parallel, the position command and the speed command are respectively processed by PI, and then are superposed with commands generated by the speed feedforward control and the acceleration feedforward control to form a current command, and the current command is processed by PI to control the servo motor. The position loop and the speed loop are arranged in parallel, speed feedforward and acceleration feedforward control are added simultaneously, and a plurality of state variables are controlled simultaneously, so that the stability of position and speed control is improved, the dynamic response performance is improved, and the position error is reduced.

The detailed structure of the control loop is shown in FIG. 7, and three sets of speed observer coefficients f in the control loop₁、f₂And f₃The system can be adjusted in real time according to the current speed change, and the response speed and the motion stability of the system under different speed conditions are ensured. And different control gains are adopted according to different motion states by using a control gain switching strategy, so that the following accuracy in static, acceleration and deceleration and uniform-speed running states is ensured.

Position proportional gain K in position loop_pHas the effect of reducing the position control error, the position integral gain K_iIs as K_pCan eliminate static error and speed proportional gain K_pvHas the effect of increasing the speed control accuracy, the speed integral gain K_ivAlso as K_pvTo eliminate speed control errors.

Compared with single feedback control, the feedforward control is more timely and effective, the interference can be predicted in advance according to the change of the interference, the real-time compensation is carried out on the interference, and the speed feedforward gain K_vffSum acceleration feedforward gain K_affThe response speed of the system can be increased, and the time for reaching the steady state is shortened. Due to the integral gain K of the velocity_ivAnd position integral gain K_iThe energy of the integration action is accumulated, and most of the feedback of the current set point in the current loop is responded to the integral terms from the position loop and the speed loop. During variable-speed motion, K_vffAnd K_affThe function of quickly reducing the integral term is achieved, so that the response of the system can be accelerated, and the stable state can be reached more quickly. Meanwhile, the feed-forward function can perform certain prejudgment on the current change of the motor when the system is accelerated and decelerated, and the response of the system can be further accelerated.

The calculation formula of the position, the speed loop and the current loop is as follows:

PWM_motor＝K_pc·I_error+K_ic·∫I_error(2)

wherein, I_setSetting a motor current value; i is_offsetIs a current bias; p_errorIs a position error; v_errorIs the speed error; PWM_motorThe PWM duty ratio value of the motor is obtained; k_pcProportional gain of current loop; k_icThe gain is integrated for the current loop.

The Z-axis moving platform 140 is used for measuring the repeated positioning error of the servo controller, the C-shaped mechanical arm 160 is provided with an inclined micro-optical system 161, a standard resolution plate is fixedly arranged at the object space focus of the inclined micro-optical system 161, the distance between adjacent scales on the standard resolution plate is 2 micrometers, and the position of an original point is calibrated on the touch screen. The servo controller controls the tilting microscopic optical system 161 to move up and down to be far away from the standard resolution board, the approaching experiment is respectively carried out from the upper direction and the lower direction, the experiment is repeated for 20 times, and the grating interval of the target position deviating from the original point at each time is read on the touch screen. The maximum deviation is 3 grating intervals in 20 uplink experiments, the maximum positioning error is about 6 microns, the maximum deviation is 5 grating intervals in 20 downlink experiments, the maximum positioning error is about-10 microns, and the repeated positioning error is mainly the self error of the Z-axis moving platform 140. The processing error of the reticle of the resolution plate and the manual reading error are considered, the repeated positioning error of the servo controller is less than +/-15 mu m, and the microscopic monitoring requirement is met.

While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.

It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.