CN110927079A - Cell culture monitoring method - Google Patents

Cell culture monitoring method Download PDF

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CN110927079A
CN110927079A CN201910387914.0A CN201910387914A CN110927079A CN 110927079 A CN110927079 A CN 110927079A CN 201910387914 A CN201910387914 A CN 201910387914A CN 110927079 A CN110927079 A CN 110927079A
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light
cell culture
sample
photodetector
sample chamber
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CN110927079B (en
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索奕双
郭强
张向平
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Jinhua Polytechnic
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Abstract

本发明涉及生命科学研究领域,一种细胞培养监测方法,细胞培养监测装置包括光源基座、九个发光二极管、孔板、九个样品室、探测器基座、九个光波导、九个光探测器、电缆和计算机,采用特殊材料制成的光波导将经过培养基的光引入光探测器,光探测灵敏度高,在不改变培养环境的条件下能够连续并量化地监测细胞培养的状态,而无需改变细胞培养环境;采用亚甲基蓝溶液来标定样品室中的溶液浓度与光探测器测得的光吸收之间的关系;采用待测的培养基来标定样品室中的酸碱度与光探测器测得的光吸收之间的关系;得到待测的细胞培养样品及培养基的浓度,同时,将九个光探测器测得的对530纳米的光的吸收的值log(I0/It)取平均,得到样品及培养基的酸碱度。

Figure 201910387914

The invention relates to the field of life science research, and a cell culture monitoring method. A cell culture monitoring device comprises a light source base, nine light-emitting diodes, an orifice plate, nine sample chambers, a detector base, nine optical waveguides, and nine optical waveguides. Detectors, cables and computers, using optical waveguides made of special materials to introduce the light passing through the culture medium into the photodetectors, the light detection sensitivity is high, and the state of cell culture can be continuously and quantitatively monitored without changing the culture environment, Without changing the cell culture environment; use methylene blue solution to calibrate the relationship between the solution concentration in the sample chamber and the light absorption measured by the photodetector; use the medium to be tested to calibrate the pH in the sample chamber and the photodetector measurement The relationship between the obtained light absorption; the concentration of the cell culture sample and the medium to be tested is obtained, and at the same time, the value of the absorption of the light at 530 nm measured by the nine photodetectors is log(I 0 /I t ) Take the average to obtain the pH of the sample and medium.

Figure 201910387914

Description

Cell culture monitoring method
Technical Field
The invention relates to the field of life science research, in particular to a cell culture monitoring method capable of continuously and quantitatively monitoring a cell culture state.
Background
In cell culture and regenerative medicine research, continuous monitoring of cell culture is very important, and conditions of the culture medium such as temperature, humidity, pH value and the like are usually required to be adjusted, so that monitoring of the culture medium is critical to cell culture, and the state of the culture medium in a cell culture chamber is usually controlled by exchanging with fresh culture medium or monitoring pH value, but these methods depend on the experience and ability of laboratory personnel, so that it is very important to develop a device for continuous monitoring and quantitative estimation of the culture medium and the cell state. The prior art methods for evaluating the state of cell culture by using devices include the yield estimation of products such as proteins and amino acids and the recovery of the culture medium, and the corresponding state is usually determined by measuring the light absorption of each species in the culture medium, but because the light detection efficiency in the detection experiment is low, the cell culture chamber needs to be taken out of the incubator, and the cover of the cell culture dish needs to be opened, so that the pollution is easily introduced and the experimental steps are complicated, and the cell culture monitoring method can solve the problems.
Disclosure of Invention
In order to solve the above problems, the method of the present invention uses a special optical waveguide to guide light passing through the culture medium to a photodetector, and can monitor the state of cell culture continuously and quantitatively without changing the culture environment.
The technical scheme adopted by the invention is as follows:
the cell culture monitoring device comprises a light source base, nine light-emitting diodes, a pore plate, nine sample chambers, a detector base, nine optical waveguides, nine optical detectors, a cable and a computer, wherein xy z is a three-dimensional space coordinate system, the light-emitting diodes emit white light, a cell culture sample is arranged in each sample chamber, the light source base is positioned at a position 12 mm above the pore plate, the pore plate is provided with nine sample chambers arranged in a 3 x 3 matrix form, the nine light-emitting diodes are arranged on the lower surface of the light source base in a 3 x 3 matrix form, the nine light-emitting diodes are respectively and correspondingly positioned right above the nine sample chambers, the nine sample chambers are all cylindrical pits with upward openings, the detector base is connected below the pore plate, the detector base is provided with nine optical waveguides arranged in a 3 x 3 matrix form, the nine optical waveguides are respectively and correspondingly positioned right below the nine sample chambers, the side surfaces of the optical waveguides are provided with light absorption coatings I with the thickness of 200 micrometers, each light absorption coating I is composed of a mixture of carbon black particles and a polysiloxane compound, the outer sides of the light absorption coatings I are covered with light absorption coatings II with the thickness of 500 micrometers, each light absorption coating II is composed of a mixture of a multi-walled carbon nanotube and polydimethylsiloxane, the lower surfaces of the nine optical waveguides are correspondingly provided with optical detectors, each optical detector is provided with a color sensor, the optical detectors are sensitive to light with the wavelengths of 620 nanometers, 530 nanometers, 460 nanometers and 860 nanometers, the nine optical detectors are all connected with a computer through cables, and the optical detectors can output detection results to the computer through 16-digit digital signals; the light source base is a square plastic sheet with the thickness of 2 mm and the side length of 30 mm, the light emitting diode is a cylinder with the height of 6 mm and the diameter of the bottom surface of 2 mm, the pore plate is a square glass sheet with the thickness of 5 mm and the side length of 30 mm, the diameter of the bottom surface of the sample chamber is 5 mm and the depth of 4 mm, the detector base is a square glass sheet with the thickness of 12 mm and the side length of 30 mm, the light guide is a cylinder with the height of 7 mm and the diameter of the bottom surface of 1 mm, the light guide is made of a transparent polysiloxane compound, the diameter of carbon black particles is 200 nm, the mass ratio of the carbon black particles of the light absorption coating I to the polysiloxane compound is 9: 1, and the mass ratio of the multi-walled carbon nano tube of the light absorption coating II to the polydimethylsiloxane is 5.
The cell culture monitoring method comprises the following steps:
step one, calibrating the relation between the solution concentration in the sample chamber and the light absorption measured by the light detector by adopting a methylene blue solution:
respectively placing the aqueous solutions of methylene blue with the concentrations of 2 micromolar, 4 micromolar, 8 micromolar, 15 micromolar, 20 micromolar, 25 micromolar, 30 micromolar, 50 micromolar and 70 micromolar in each sample chamber, starting the light-emitting diode, recording the light intensity and wavelength information of the solutions passing through each sample chamber by using a light detector, and calculating to obtain the absorption log (I) of the solutions with the concentrations to light0/It) In which I0Is the initial light intensity of light with the wavelength of 620 nanometers in white light emitted by the light emitting diode, ItThe light intensity of light which is measured by the light detector and is 620 nanometers, penetrates through the cell culture sample and enters the light detector;
step two, calibrating the relationship between the pH value in the sample chamber and the light absorption measured by the light detector by using a culture medium to be measured:
the known pH values of 7.0, 7.2, 7.4, 7.8, 8.0, 8.4, 8.8, 9.0 and 9.4 to be measuredThe culture medium is placed in each sample chamber, the light emitting diode is started, the light detector is adopted to record the light intensity and wavelength information of the culture medium passing through each sample chamber, and the absorption value log (I) of the culture medium with different pH values to light is calculated0/It) In which I0Is the initial light intensity, I, of light with the wavelength of 530 nanometers in white light emitted by a light emitting diodetThe light intensity of light transmitted through the cell culture sample and entering the light detector at 530 nm as measured by the light detector;
mixing nine identical cell culture samples to be detected with a fresh culture medium, placing the mixture into nine sample chambers, starting the light-emitting diodes, recording the light intensity and wavelength information of the culture medium passing through each sample chamber by using the light detectors, and measuring the value log (I log) of the absorption of light of 620 nanometers by using the nine light detectors0/It) Averaging to obtain the absorption of the cell culture sample to be detected to the light of 620 nanometers, and comparing with the relation between the solution concentration in the sample chamber obtained in the step one and the light absorption measured by the optical detector to finally obtain the concentration of the cell culture sample to be detected and the concentration of the culture medium;
step four, the step four and the step three are carried out simultaneously, and the value log (I) of the absorption of the light with the wavelength of 530 nanometers measured by nine photodetectors0/It) And averaging to obtain the absorption of the cell culture sample to be detected to 530 nm of light, and comparing with the relationship between the pH value in the sample chamber obtained in the step two and the light absorption measured by the light detector to finally obtain the pH value of the cell culture sample to be detected and the pH value of the culture medium.
The invention has the beneficial effects that:
the method of the invention adopts the optical waveguide made of special materials, has high light detection sensitivity, and can continuously and quantitatively monitor the state of cell culture without changing the cell culture environment.
Drawings
The following is further illustrated in connection with the figures of the present invention:
FIG. 1 is a schematic of the present invention;
FIG. 2 is a perspective view of a light source base, aperture plate, and detector base.
In the figure, 1, a light source base, 2, a light emitting diode, 3, a pore plate, 4, a sample chamber, 5, a detector base, 6, an optical waveguide and 7, a light detector are arranged.
Detailed Description
FIG. 1 is a schematic view of the present invention, FIG. 2 is a schematic perspective view of a light source base, a pore plate and a detector base, which includes a light source base (1), nine light emitting diodes (2), a pore plate (3), nine sample chambers (4), a detector base (5), nine optical waveguides (6), nine photodetectors (7), a cable and a computer, xy z is a three-dimensional space coordinate system, the light emitting diodes (2) emit white light, a cell culture sample is disposed in the sample chambers (4), the light source base (1) is a square plastic sheet with a thickness of 2 mm and a side length of 30 mm, the light source base (1) is disposed at 12 mm above the pore plate (3), the pore plate (3) is a square glass sheet with a thickness of 5 mm and a side length of 30 mm, the pore plate (3) has nine sample chambers (4) arranged in a matrix of 3 × 3, the nine light emitting diodes (2) are mounted on the lower surface of the light source base (1) in a matrix of 3 × 3, nine light-emitting diodes (2) are respectively and correspondingly positioned right above nine sample chambers (4), the light-emitting diodes (2) are cylinders with the height of 6 mm and the diameter of the bottom surface of 2 mm, the nine sample chambers (4) are all cylindrical pits with upward openings, the diameter of the bottom surface of each sample chamber (4) is 5 mm and the depth of 4 mm, a detector base (5) is a square glass sheet with the thickness of 12 mm and the side length of 30 mm, the detector base (5) is connected below a pore plate (3), nine optical waveguides (6) arranged in a 3 x 3 matrix form are arranged on the detector base (5), the optical waveguides (6) are cylinders with the height of 7 mm and the diameter of the bottom surface of 1 mm, the optical waveguides (6) are made of a transparent polysiloxane compound, the nine optical waveguides (6) are respectively and correspondingly positioned right below the nine sample chambers (4), the side surfaces of the optical waveguides (6) are respectively provided with a light absorption coating I with the thickness of 200 microns, the light absorption coating I is composed of a mixture of carbon black particles and polysiloxane compounds, the diameter of the carbon black particles is 200 nanometers, and the mass ratio of the carbon black particles to the polysiloxane compounds of the light absorption coating I is 9: 1; the outer side of the light absorption coating I is covered with a light absorption coating II with the thickness of 500 micrometers, the light absorption coating II is composed of a mixture of multi-walled carbon nanotubes and polydimethylsiloxane, the mass ratio of the multi-walled carbon nanotubes to the polydimethylsiloxane of the light absorption coating II is 5: 1, a light detector (7) is correspondingly installed below nine optical waveguides (6), the light detector (7) is provided with a color sensor, so that the light detector (7) is sensitive to light with the wavelengths of 620 nanometers, 530 nanometers, 460 nanometers and 860 nanometers, all the nine light detectors (7) are connected with a computer through cables, and the light detector (7) can output detection results to the computer through 16-bit digital signals.
The principle of the invention with higher light detection sensitivity is as follows: the white light emitted by the light emitting diode (2) irradiates on the cell culture sample in the sample chamber (4) and is absorbed in different degrees at certain wavelengths, a part of light which is not absorbed by the cell culture sample passes through the bottom surface of the pore plate (3) and is transmitted into the optical waveguide (6), the light is reflected and scattered at the interface between the cell culture sample and the bottom surface of the sample chamber (4) and the interface between the bottom surface of the pore plate (3) and the optical waveguide (6) to generate stray light, so that the signal-to-noise ratio of the optical detector (7) is influenced, because the light absorbing coating I and the light absorbing coating II on the side surface of the optical waveguide (6) have strong light absorption, therefore, most of the stray light can be coated by the light absorption layer, and only the light which propagates along the negative y direction can enter the light detector (7) through the optical waveguide (6), so that the light detection sensitivity is improved.
The cell culture monitoring device comprises a light source base (1), nine light emitting diodes (2), a pore plate (3), nine sample chambers (4), a detector base (5), nine optical waveguides (6), nine optical detectors (7), a cable and a computer, wherein xyz is a three-dimensional space coordinate system, the light emitting diodes (2) emit white light, cell culture samples are arranged in the sample chambers (4), the light source base (1) is positioned at 12 mm above the pore plate (3), the pore plate (3) is provided with nine sample chambers (4) arranged in a 3 x 3 matrix form, the nine light emitting diodes (2) are arranged on the lower surface of the light source base (1) in a 3 x 3 matrix form, the nine light emitting diodes (2) are respectively and correspondingly positioned right above the nine sample chambers (4), the nine sample chambers (4) are cylindrical pits with upward openings, the detector base (5) is connected below the pore plate (3), nine optical waveguides (6) arranged in a 3 x 3 matrix form are arranged on the detector base (5), the nine optical waveguides (6) are respectively and correspondingly positioned right below the nine sample chambers (4), the side surfaces of the optical waveguides (6) are respectively provided with a light absorption coating I with the thickness of 200 micrometers, the light absorption coating I is composed of a mixture of carbon black particles and polysiloxane compounds, the outer side of the light absorption coating I is covered with a light absorption coating II with the thickness of 500 micrometers, the light absorption coating II is composed of a mixture of multi-walled carbon nanotubes and polydimethylsiloxane, a light detector (7) is correspondingly arranged below the nine optical waveguides (6), and the light detector (7) is provided with a color sensor, the light detectors (7) are sensitive to light with wavelengths of 620 nm, 530 nm, 460 nm and 860 nm, nine light detectors (7) are all connected with a computer through cables, and the light detectors (7) can output detection results of 16-bit digital signals to the computer; the light source base (1) is a square plastic sheet with the thickness of 2 mm and the side length of 30 mm, the light emitting diode (2) is a cylinder with the height of 6 mm and the diameter of the bottom surface of 2 mm, the pore plate (3) is a square glass sheet with the thickness of 5 mm and the side length of 30 mm, the diameter of the bottom surface of the sample chamber (4) is 5 mm, the depth is 4 mm, the detector base (5) is a square glass sheet with the thickness of 12 mm and the side length of 30 mm, the optical waveguide (6) is a cylinder with the height of 7 mm and the diameter of the bottom surface of 1 mm, the optical waveguide (6) is made of a transparent polysiloxane compound, the diameter of carbon black particles is 200 nm, the mass ratio of the carbon black particles to the polysiloxane compound of the light absorption coating I is 9: 1, and the mass ratio of multi-wall carbon nano tubes to polydimethylsiloxane of the light absorption coating II is 5: 1.
The cell culture monitoring method comprises the following steps:
step one, calibrating the relation between the concentration of the solution in the sample chamber (4) and the light absorption measured by the light detector (7) by adopting a methylene blue solution:
respectively adopting 2 micromolar, 4 micromolar, 8 micromolar, 15 micromolar, 20 micromolar, 25 micromolar, 30 micromolar, 50 micromolar and 70 micromolar aqueous solutions to be placed in each sample chamber (4), starting the light-emitting diode (2), recording the light intensity and wavelength information of the solution passing through each sample chamber (4) by using a light detector (7), and calculating to obtain the value log (I) of the absorption of the solution with each concentration to light0/It) In which I0To emit lightInitial intensity, I, of light with a wavelength of 620 nm in the white light emitted by the diode (2)tThe light intensity of the light of 620 nm measured by the light detector (7) which penetrates through the cell culture sample and enters the light detector (7);
step two, calibrating the relationship between the pH value in the sample chamber (4) and the light absorption measured by the light detector (7) by using a culture medium to be measured:
placing culture media to be detected with known pH values of 7.0, 7.2, 7.4, 7.8, 8.0, 8.4, 8.8, 9.0 and 9.4 into each sample chamber (4), starting the light emitting diode (2), recording the light intensity and wavelength information of the culture media passing through each sample chamber (4) by using the light detector (7), and calculating to obtain the absorption log (I) of the culture media with different pH values to light0/It) In which I0Is the initial light intensity, I, of light with the wavelength of 530 nanometers in the white light emitted by the light emitting diode (2)tThe light intensity of light measured by the light detector (7) at 530 nm transmitted through the cell culture sample and entered the light detector (7);
mixing nine same cell culture samples to be detected with a fresh culture medium, placing the mixture into nine sample chambers (4), starting the light-emitting diodes (2), recording the light intensity and wavelength information of the culture medium in each sample chamber (4) by using the light detectors (7), and measuring the absorption value log (I) of light with 620 nanometers by using the nine light detectors (7)0/It) Averaging to obtain the absorption of the cell culture sample to be detected to the light of 620 nanometers, and comparing with the relation between the solution concentration in the sample chamber (4) obtained in the step one and the light absorption measured by the light detector (7), so as to finally obtain the concentrations of the cell culture sample to be detected and the culture medium;
step four, the step four and the step three are carried out simultaneously, and the value log (I) of the absorption of the light with the wavelength of 530 nanometers measured by the nine photodetectors (7) is measured0/It) And averaging to obtain the absorption of the cell culture sample to be detected to the light of 530 nanometers, and comparing the absorption with the relation between the pH value in the sample room (4) obtained in the step two and the light absorption measured by the light detector (7) to finally obtain the pH value of the cell culture sample to be detected and the pH value of the culture medium.
The method can measure the pH value change of the culture medium in real time under the condition of not changing the culture environment, and can detect the concentration change of the culture medium with high precision.

Claims (1)

1.一种细胞培养监测方法,细胞培养监测装置包括光源基座(1)、九个发光二极管(2)、孔板(3)、九个样品室(4)、探测器基座(5)、九个光波导(6)、九个光探测器(7)、电缆和计算机,xyz为三维空间坐标系,发光二极管(2)发出白光,样品室(4)内具有细胞培养样品,光源基座(1)位于孔板(3)上方的12毫米处,孔板(3)上具有九个以3×3矩阵形式排列的样品室(4),九个发光二极管(2)以3×3矩阵形式排列地安装于光源基座(1)的下表面,九个发光二极管(2)分别对应位于九个样品室(4)的正上方,九个样品室(4)均是开口向上的圆柱形坑,探测器基座(5)连接于孔板(3)下面,探测器基座(5)上具有九个以3×3矩阵形式排列的光波导(6),九个光波导(6)分别对应位于九个样品室(4)的正下方,光波导(6)的侧面均具有厚度为200微米的光吸收涂层I,光吸收涂层I由碳黑颗粒与聚硅氧烷化合物的混合物组成,光吸收涂层I的外侧覆盖有厚度为500微米的光吸收涂层II,光吸收涂层II由多壁碳纳米管与聚二甲硅氧烷的混合物组成,九个光波导(6)下面均对应安装有一个光探测器(7),光探测器(7)具有色彩传感器,使得光探测器(7)对波长为620纳米、530纳米、460纳米和860纳米的光敏感,九个光探测器(7)均电缆连接计算机,光探测器(7)能够以16位数字信号探测结果输出至计算机;光源基座(1)是厚度为2毫米、边长为30毫米的正方形塑料片,发光二极管(2)的外形是高度为6毫米、底面直径为2毫米的圆柱体,孔板(3)是厚度为5毫米、边长为30毫米的正方形玻璃片,样品室(4)的底面直径为5毫米、深度为4毫米,探测器基座(5)是厚度为12毫米、边长为30毫米的正方形玻璃片,光波导(6)的外形是高度为7毫米、底面直径为1毫米的圆柱体,光波导(6)由透光的聚硅氧烷化合物制成,碳黑颗粒的直径为200纳米,光吸收涂层I的碳黑颗粒与聚硅氧烷化合物的质量比例为9∶1,光吸收涂层II的多壁碳纳米管与聚二甲硅氧烷的质量比例为5∶1,1. A cell culture monitoring method, the cell culture monitoring device comprising a light source base (1), nine light-emitting diodes (2), a well plate (3), nine sample chambers (4), and a detector base (5) , nine optical waveguides (6), nine photodetectors (7), cables and a computer, xyz is a three-dimensional space coordinate system, the light-emitting diode (2) emits white light, the sample chamber (4) has a cell culture sample, and the light source base The seat (1) is located 12 mm above the orifice plate (3) with nine sample chambers (4) arranged in a 3×3 matrix, nine light-emitting diodes (2) in a 3×3 matrix. The nine light-emitting diodes (2) are arranged in a matrix form on the lower surface of the light source base (1), and the nine light-emitting diodes (2) are respectively located directly above the nine sample chambers (4), and the nine sample chambers (4) are all cylinders with upward openings. The detector base (5) is connected to the bottom of the orifice plate (3), the detector base (5) has nine optical waveguides (6) arranged in a 3×3 matrix, and the nine optical waveguides (6 ) are respectively located directly under the nine sample chambers (4), and the sides of the optical waveguide (6) are provided with a light-absorbing coating I with a thickness of 200 microns, and the light-absorbing coating I is composed of carbon black particles and polysiloxane compounds. The outer side of the light-absorbing coating I is covered with a light-absorbing coating II with a thickness of 500 microns, and the light-absorbing coating II is composed of a mixture of multi-walled carbon nanotubes and polydimethylsiloxane. Nine optical waveguides (6) a photodetector (7) is correspondingly installed below, and the photodetector (7) has a color sensor, so that the photodetector (7) is sensitive to light with wavelengths of 620 nm, 530 nm, 460 nm and 860 nm , the nine photodetectors (7) are all connected to the computer by cables, and the photodetectors (7) can output the detection results of 16-bit digital signals to the computer; the light source base (1) is 2 mm thick and 30 mm long A square plastic sheet, the shape of the light emitting diode (2) is a cylinder with a height of 6 mm and a bottom diameter of 2 mm, the orifice plate (3) is a square glass sheet with a thickness of 5 mm and a side length of 30 mm, the sample chamber ( 4) The diameter of the bottom surface is 5 mm and the depth is 4 mm, the detector base (5) is a square glass sheet with a thickness of 12 mm and a side length of 30 mm, and the shape of the optical waveguide (6) is 7 mm in height, A cylinder with a bottom diameter of 1 mm, the optical waveguide (6) is made of a light-transmitting polysiloxane compound, the diameter of the carbon black particles is 200 nanometers, and the carbon black particles of the light absorbing coating I and the polysiloxane compound The mass ratio of MWNT is 9:1, the mass ratio of multi-walled carbon nanotubes and polydimethylsiloxane in the light absorbing coating II is 5:1, 其特征是:所述一种细胞培养监测方法的步骤为:It is characterized in that: the steps of the cell culture monitoring method are: 步骤一,采用亚甲基蓝溶液来标定样品室(4)中的溶液浓度与光探测器(7)测得的光吸收之间的关系:Step 1, use methylene blue solution to calibrate the relationship between the solution concentration in the sample chamber (4) and the light absorption measured by the photodetector (7): 分别采用浓度为2、4、8、15、20、25、30、50和70微摩尔的亚甲基蓝的水溶液置于各样品室(4)内,开启发光二极管(2),采用光探测器(7)记录通过各样品室(4)内溶液的光强及波长信息,并计算得到各浓度的溶液对光的吸收log(I0/It),其中I0为发光二极管(2)发出的白光中波长为620纳米的光的初始光强,It为光探测器(7)测得的620纳米的光透过细胞培养样品并进入光探测器(7)中的光的光强;The aqueous solutions of methylene blue with concentrations of 2, 4, 8, 15, 20, 25, 30, 50 and 70 micromoles were respectively placed in each sample chamber (4), the light-emitting diode (2) was turned on, and a photodetector (7) was used. ) record the light intensity and wavelength information passing through the solution in each sample chamber (4), and calculate the light absorption log(I 0 /I t ) of the solution of each concentration, where I 0 is the white light emitted by the light-emitting diode (2) The initial light intensity of the light with the medium wavelength of 620 nm, I t is the light intensity of the light at 620 nm measured by the photodetector (7), which transmits the cell culture sample and enters the photodetector (7); 步骤二,采用待测的培养基来标定样品室(4)中的酸碱度与光探测器(7)测得的光吸收之间的关系:Step 2, using the culture medium to be tested to calibrate the relationship between the pH in the sample chamber (4) and the light absorption measured by the photodetector (7): 将已知酸碱度为7.0、7.2、7.4、7.8、8.0、8.4、8.8、9.0和9.4的待测的培养基置于各样品室(4)内,开启发光二极管(2),采用光探测器(7)记录通过各样品室(4)内培养基的光强及波长信息,并计算得到各不同酸碱度的培养基对光的吸收的值log(I0/It),其中I0为发光二极管(2)发出的自光中波长为530纳米的光的初始光强,It为光探测器(7)测得的530纳米的光透过细胞培养样品并进入光探测器(7)中的光的光强;Place the medium to be tested with known pH values of 7.0, 7.2, 7.4, 7.8, 8.0, 8.4, 8.8, 9.0 and 9.4 in each sample chamber (4), turn on the light-emitting diode (2), and use a photodetector ( 7) Record the light intensity and wavelength information passing through the medium in each sample chamber (4), and calculate the value log(I 0 /I t ) of the light absorption of the medium with different pH, where I 0 is the light-emitting diode (2) The initial light intensity of the light with a wavelength of 530 nm in the emitted self-light, I t is the light of 530 nm measured by the photodetector (7) that penetrates the cell culture sample and enters the photodetector (7) the intensity of light; 步骤三,将九份相同的待测的细胞培养样品与新鲜的培养基混合后,置于九个样品室(4)内,开启发光二极管(2),采用光探测器(7)记录通过各样品室(4)内培养基的光强及波长信息,将九个光探测器(7)测得的对620纳米的光的吸收的值log(I0/It)取平均,以得到待测的细胞培养样品对620纳米的光的吸收,并与步骤一中所得的样品室(4)中的溶液浓度与光探测器(7)测得的光吸收之间的关系进行比较,最终得到待测的细胞培养样品及培养基的浓度;In step 3, after mixing nine identical cell culture samples to be tested with fresh culture medium, they are placed in nine sample chambers (4), light-emitting diodes (2) are turned on, and photodetectors (7) are used to record the passage through each cell. For the light intensity and wavelength information of the culture medium in the sample chamber (4), the value log(I 0 /I t ) of the absorption of light at 620 nanometers measured by the nine photodetectors (7) is averaged to obtain the The measured cell culture sample absorbs light at 620 nm, and compares it with the relationship between the solution concentration in the sample chamber (4) obtained in step 1 and the light absorption measured by the photodetector (7), and finally obtains The concentration of the cell culture sample to be tested and the culture medium; 步骤四,步骤四与步骤三同时进行,将九个光探测器(7)测得的对530纳米的光的吸收的值log(I0/It)取平均,以得到待测的细胞培养样品对530纳米的光的吸收,并与步骤二中所得的样品室(4)中的酸碱度与光探测器(7)测得的光吸收之间的关系进行比较,最终得到待测的细胞培养样品及培养基的酸碱度。Step 4, step 4 and step 3 are carried out simultaneously, and the value log(I 0 /I t ) of the absorption of light at 530 nanometers measured by the nine photodetectors (7) is averaged to obtain the cell culture to be tested. The absorption of light at 530 nanometers by the sample is compared with the relationship between the pH in the sample chamber (4) obtained in step 2 and the light absorption measured by the photodetector (7), and finally the cell culture to be tested is obtained. pH of samples and media.
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