CN211057126U

CN211057126U - Cell culture monitoring devices

Info

Publication number: CN211057126U
Application number: CN201920663979.9U
Authority: CN
Inventors: 张向平; 范晓雯; 方晓华
Original assignee: Jinhua Polytechnic
Current assignee: Jinhua Polytechnic
Priority date: 2019-05-02
Filing date: 2019-05-02
Publication date: 2020-07-21
Anticipated expiration: 2029-05-02

Abstract

The utility model relates to a life science research field, a cell culture monitoring devices, including the light source base, nine emitting diode, the orifice plate, nine sample rooms, the detector base, nine optical waveguides, nine optical detector, cable and computer, the light source base is located the 12 millimeters department of orifice plate top, nine sample rooms of arranging with 3 × 3 matrix form have on the orifice plate, nine emitting diode are installed in the lower surface of light source base with 3 × 3 matrix form is arranged, and correspond respectively and be located nine sample rooms directly over, nine sample rooms all are the ascending cylindrical hole of opening, the detector pedestal connection is below the orifice plate, nine optical waveguides that arrange with 3 × 3 matrix form have on the detector base, nine optical waveguides correspond respectively and are located nine sample rooms under, the side of optical waveguide all has light absorption coating I that thickness is 200 microns, light absorption coating I's the outside covers has light absorption coating II that thickness is 500 microns, the equal cable junction computer of optical detector.

Description

Cell culture monitoring devices

Technical Field

The utility model belongs to the technical field of the life science research and specifically relates to a can monitor cell culture monitoring devices of cell culture state in succession and quantization.

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 device can solve the problems.

SUMMERY OF THE UTILITY MODEL

In order to solve the above problems, the present invention provides a device for filtering light passing through a culture medium by means of a special optical waveguide, which can monitor the state of cell culture continuously and quantitatively without changing the cell culture environment.

The utility model adopts the technical proposal that:

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 xyz 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 located 12 mm above the pore plate, the pore plate is provided with nine sample chambers arranged in a 3 × 3 matrix form, the nine light-emitting diodes are arranged on the lower surface of the light source base in a 3 × 3 matrix form and are respectively located right above the nine sample chambers, the nine sample chambers are all cylindrical with upward openings, the detector base is connected to the lower surface of the pore plate, the detector base is provided with nine optical waveguides arranged in a 3 × 3 matrix form, the nine optical waveguides are respectively located right below the nine sample chambers correspondingly, the side surfaces of the optical waveguides are all provided with an optical absorption coating I with the thickness of 200 micrometers, the optical absorption coating I is composed of a mixture of carbon black particles and polysiloxane compounds, the outer side of the optical absorption coating II is covered with the thickness of 500 micrometers, the optical waveguide, the optical absorption coating II is composed of a polysiloxane compound, the polysiloxane compound is a polysiloxane compound, the polysiloxane compound is a polysiloxane compound, the polysiloxane.

The cell culture monitoring experiment carried out by adopting the cell culture monitoring device 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:

placing aqueous solutions of methylene blue with concentrations of 2, 4, 8, 15, 20, 25, 30, 50 and 70 micromoles in each sample chamber, turning on the light emitting diode, and adoptingThe light detector records the light intensity and wavelength information of the solution passing through each sample chamber, and calculates the absorption value log (I) of the solution with each concentration to the light₀/I_t) In which I₀Is the initial light intensity of light with the wavelength of 620 nanometers in white light emitted by the light emitting diode, I_tThe 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:

placing culture 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, starting light emitting diodes, recording light intensity and wavelength information of the culture medium passing through each sample chamber by using a light detector, and calculating to obtain absorption log (I) of the culture medium with different pH values to light₀/I_t) In which I₀Is the initial light intensity, I, of light with the wavelength of 530 nanometers in white light emitted by a light emitting diode_tThe 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/I_t) 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/I_t) Averaging to obtain the absorption of 530 nm light by the cell culture sample to be tested, and comparing with the pH value in the sample chamber obtained in the step two and the light absorption measured by the photodetectorAnd comparing the relationship between the two groups of the culture medium to obtain the pH value of the cell culture sample to be detected and the culture medium.

The utility model has the advantages that:

the utility model discloses the device can real-time supervision cell cultivate the environment, can measure the pH valve change of culture medium in real time under the condition that does not change the cultivation environment to can come the concentration change of surveying the culture medium high-precision through methylene blue's absorption.

Drawings

The following is further illustrated in connection with the figures of the present invention:

FIG. 1 is a schematic view 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 view of a light source base, a pore plate and a detector base, including a light source base (1), nine light emitting diodes (2), a pore plate (3), nine sample chambers (4), a detector base (5), nine light waveguides (6), nine photodetectors (7), a cable and a computer, xyz is a three-dimensional space coordinate system, the light emitting diodes (2) emit white light, a cell culture sample is provided in the sample chamber (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 located 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 3 × matrix form, the nine light emitting diodes (2) are arranged on the lower surface of the light source base (1) in a 3 × matrix form, the nine light emitting diodes (2) are respectively located above the sample chambers (4), the sample chambers are connected to the bottom surface of the polysiloxane base (1) by a polysiloxane light waveguide (3), the polysiloxane light waveguide (2) and the polysiloxane (2) are connected to the polysiloxane light waveguide (2), the polysiloxane light waveguide (2) by a polysiloxane light absorbing carbon black, the polysiloxane light absorbing carbon black carbon.

The utility model discloses the principle that the device has higher light detection sensitivity is: 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 × 3 matrix form, the nine light-emitting diodes (2) are arranged on the lower surface of the light source base (1) in a 3 × 3 matrix form, the nine light-emitting diodes (2) are respectively positioned right above the nine sample chambers (4), the nine sample chambers (4) are all cylindrical with an opening upwards, the detector base (5) is connected to the lower surface of the detector base (3), the detector base (5) is provided with nine optical waveguides (6) arranged in a 3 × 3 matrix form, the optical waveguides (6) are respectively positioned right above the nine sample chambers (4) and are positioned on the cylindrical carbon black, the bottom of a square, the polysiloxane substrate (2) and a polysiloxane-carbon black.

The utility model discloses the device has special optical waveguide, and the light detection sensitivity is high, can monitor cell culture's state in succession and quantization under the condition that does not change the environment of cultivateing.

Claims

1. A 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, and a cell culture sample is arranged in each sample chamber (4), the cell culture monitoring device is characterized in that 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 × 3 matrix form, the nine light emitting diodes (2) are arranged on the lower surface of the light source base (1) in a 3 × 3 matrix form, the nine light emitting diodes (2) are respectively positioned right above the nine sample chambers (4), the nine sample chambers (4) are all cylindrical pits with upward openings, the detector base (5) is connected to the lower surface of the pore plate (3), the detector base (5) is provided with nine optical waveguides (6) arranged in a 3 × 3 matrix form, the nine sample chambers (6) are respectively positioned right above the optical cables, the optical detectors (6) and are respectively positioned below the optical detectors (I) and connected with a light absorption coating layer which is used for absorbing light, the light sensors (I) and the light detectors (7) and the light sensors are respectively arranged on the lateral surfaces of which are connected with the light sensors, the light sensors are connected with the light absorption coating layers, the light sensors are.

2. A cell culture monitoring device according to claim 1, wherein: the light source base (1) is a square plastic sheet with a thickness of 2 mm and a side length of 30 mm.

3. A cell culture monitoring device according to claim 1, wherein: the light emitting diode (2) has a cylindrical shape with a height of 6 mm and a diameter of 2 mm at the bottom.

4. A cell culture monitoring device according to claim 1, wherein: the perforated plate (3) is a square glass sheet with a thickness of 5 mm and a side length of 30 mm.

5. A cell culture monitoring device according to claim 1, wherein: the diameter of the bottom surface of the sample chamber (4) is 5 mm, and the depth is 4 mm.

6. A cell culture monitoring device according to claim 1, wherein: the detector base (5) is a square glass plate with a thickness of 12 mm and a side length of 30 mm.

7. A cell culture monitoring device according to claim 1, wherein: the optical waveguide (6) has a cylindrical shape with a height of 7 mm and a diameter of 1 mm at the bottom, and the optical waveguide (6) is made of a light-transmitting silicone compound.

8. A cell culture monitoring device according to claim 1, wherein: the diameter of the carbon black particles was 200 nm.