CN118421456A - Automatic change organoid culture monitoring platform - Google Patents
Automatic change organoid culture monitoring platform Download PDFInfo
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- CN118421456A CN118421456A CN202410520726.1A CN202410520726A CN118421456A CN 118421456 A CN118421456 A CN 118421456A CN 202410520726 A CN202410520726 A CN 202410520726A CN 118421456 A CN118421456 A CN 118421456A
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
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- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
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- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
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Abstract
The invention discloses an automatic organoid culture monitoring platform, which comprises a microfluidic system, a culture system, a sensing system and a high content system; the microfluidic system comprises a sealed shell, an inner groove for containing culture solution, an outer groove for containing waste liquid and the like; the culture system comprises a plurality of culture cells arranged in the culture tank, culture holes arranged in the culture cells and a first microporous filter membrane; the sensing system comprises a liquid suction device, a detection chamber, a sensing probe and a water tank, wherein the sensing probe is arranged in the detection chamber and is used for detecting various parameters of the culture solution; the high content system comprises an imaging analyzer and a computer, wherein the imaging analyzer is arranged outside the culture system and used for acquiring organoid image data in the culture hole, and the computer is connected with the imaging analyzer and the sensing system. The platform realizes the accuracy and automation of culture monitoring while ensuring the stability of the culture environment, is more suitable for the culture process of organoids, and is beneficial to improving the culture success rate and saving human resources.
Description
Technical Field
The invention relates to the technical field of organoid culture, in particular to an automatic organoid culture monitoring platform.
Background
Based on the clinical relevance of organoids over 2D culture, organoid models incorporating culture monitoring devices have become an important tool for pathophysiological research and drug screening.
In the prior art, the organoids are usually cultured in a constant-temperature carbon dioxide incubator, the culture plates are taken out for observation under a microscope, the growth state of the organoids is judged through the color of a culture medium, the morphology, the density and the like of the organoids under a mirror, and then the operations such as liquid exchange, passage, freezing storage and the like are selected. On the one hand, the process mainly depends on manual participation, errors exist in subjective experience judgment of operation time, and accuracy, intellectualization, automation and the like cannot be achieved, for example, patent CN213232314U discloses a living cell monitoring device, however, the detection device still depends on manual operation, and the method is inconvenient. On the other hand, the composition and the change of the organoid state in the cell culture liquid cannot be monitored in real time, so that a comprehensive and accurate experimental result cannot be obtained, patent CN116814431a discloses a cell culture monitoring system, which detects and processes signals through a sensor, but the system can only monitor the environmental parameters of the cell growth process, cannot monitor the volume, the number and the like of cells, is not suitable for organoid culture, and cannot realize the automation of culture operation. As patent CN116814431a discloses a cell culture monitoring system, which detects and processes signals by means of sensors, but the system can only monitor environmental parameters of the cell growth process, cannot monitor the volume, number, etc. of cells themselves, is not suitable for organoid culture, and cannot realize automation of culture operation. In addition, when the corresponding culture plate is frequently observed and searched, the incubator is opened for a long time, so that the temperature and humidity in the incubator and the concentration of carbon dioxide are unstable, even pollution is introduced, and the culture environment is damaged. The application of an automatic organoid culture real-time monitoring platform is urgent.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides an automatic organoid culture monitoring platform based on high-speed microscopy, image analysis and sensing technology, which accurately monitors the organoid growth state through high-content cell imaging and real-time monitoring of culture solution components.
In order to solve the technical problems, the invention provides the following technical scheme:
An automatic change organoid culture monitoring platform, its characterized in that: comprises a micro-fluidic system, a culture system, a sensing system and a high content system;
The microfluidic system comprises a sealed shell, an inner groove for containing culture solution, an outer groove for containing waste liquid, a liquid pump, a liquid inlet pipe and a waste liquid pipe, wherein the inner groove and the outer groove are arranged in the sealed shell, one end of the liquid inlet pipe is connected with the inner groove through a liquid inlet groove hole, the other end of the liquid inlet pipe is connected with the culture system, one end of the waste liquid pipe is connected with the outer groove through a waste liquid groove hole, the other end of the waste liquid pipe is connected with the culture system, the liquid pump is arranged in the liquid inlet groove hole, and a sensing area is arranged on the waste liquid pipe;
The culture system comprises a plurality of culture cells arranged in a culture tank, culture holes arranged in the culture cells and a first microporous filter membrane, wherein a first liquid hole and a second liquid hole are formed in the side face of the culture cells, the liquid inlet pipe and the liquid outlet pipe are respectively connected with the culture holes through the first liquid hole and the second liquid hole, an inoculation hole opened and closed through a knob is formed in the top of the culture cells and used for inoculating organoids into the culture holes, and the first microporous filter membrane is arranged on the first liquid hole and the second liquid hole and used for preventing cells from passing through;
The sensing system comprises a liquid suction device, a detection chamber, a sensing probe and a water tank, wherein one end of the liquid suction device is connected with the sensing region, the other end of the liquid suction device is connected with the detection chamber through a sensing channel, the sensing probe is arranged in the detection chamber and used for detecting various parameters of the culture solution, and the water tank is arranged at the bottom of the detection chamber;
the high content system comprises an imaging analyzer and a computer, wherein the imaging analyzer is arranged on the outer side of the culture system and used for acquiring organoid image data in a culture hole, and the computer is connected with the imaging analyzer and the sensing system.
Further, a plurality of liquid chambers are arranged in the inner groove, the liquid pump is connected with each liquid chamber, and the liquid pump is of a multi-channel structure and can draw culture liquid from different liquid chambers.
Further, the first liquid hole and the second liquid hole are respectively connected with the culture hole through an input channel and an output channel, a second microporous filter membrane with the specification of 0.22 μm is arranged at the joint, and the horizontal positions of the input channel and the output channel are higher than the culture plane at the bottom of the culture hole.
Further, the first liquid hole and the second liquid hole are respectively divided into a plurality of branches through the first multiplexer and the second multiplexer and then are connected with the culture hole.
Further, the plurality of culture holes are connected with each other through a liquid channel to form a culture cavity.
Furthermore, a micro sensing chip is embedded in the hydrogel at the bottom of the culture hole and is used for monitoring the nutrition level and the waste level in the process of medium updating.
Furthermore, the culture chamber is made of an optically transparent raw polystyrene material, and the culture hole is made of a PDMS material with good biocompatibility.
Furthermore, an environmental control device is arranged in the culture chamber, so that the temperature, humidity, oxygen and carbon dioxide concentration in the culture chamber can be controlled.
Further, the first liquid hole and the second liquid hole are respectively and tightly connected with the liquid inlet pipe and the liquid waste pipe through the liquid hole clamping groove.
Further, the sensing probe comprises a DO probe, a pH probe, a glucose probe, a lactic acid probe and a nitric oxide probe, wherein the tip of the DO probe consists of an optical fiber with the size smaller than 50 mu m, a photosensitive dye is doped into a polymer and covers the tip of the DO probe, and the concentration of the culture solution DO is detected by utilizing dynamic luminescence quenching of the photosensitive dye in the presence of oxygen based on a Stern-Volmer equation; the pH probe comprises an iridium oxide electrode, a glass film is used as an interface between the liquid to be detected and the film sealing internal buffer solution, and the pH of the culture solution is quantified by utilizing potential difference; the glucose probe is a platinum electrode, glucose oxidase, bovine serum albumin and glutaraldehyde crosslinking agent are added, and the glucose level of the culture solution is tracked in real time; the lactic acid probe contains a platinum electrode, immobilized lactic acid dehydrogenase and coenzyme as media, electrons are shuttled between the lactic acid dehydrogenase and the electrode, and the lactic acid concentration in the culture solution is quantified through oxidation current; the nitric oxide probe comprises a platinum electrode and a poly eugenol film which can selectively permeate nitric oxide, and the nitric oxide concentration of the culture solution is measured by an electrochemical method.
Compared with the prior art, the invention has the beneficial effects that: 1. the sensor, the microfluidic device and the high-content cell imager are connected through the feedback control system to form the organoid culture monitoring platform, so that a traditional incubator is replaced, a comprehensive and convenient mode of image acquisition, environment monitoring and culture solution component detection is provided in a culture environment, the culture environment is stable, meanwhile, the accuracy and automation of culture monitoring are realized, the organoid culture process is more suitable, the culture success rate is improved, human resources are saved, and the pollution risk and the adverse effects caused by culture environment change are reduced. 2. The microfluidic system can accurately quantitatively add various culture mediums with different components into different culture holes through multiplexing, so that the culture efficiency is improved. 3. The micro sensing chip is embedded at the bottom of the culture cell, so that a plurality of parameters such as DO, pH and the like can be obtained, and the functions of monitoring the nutrition level and the waste level in the process of updating the culture medium are achieved.
Drawings
FIG. 1 is a schematic diagram of an overall structure of an embodiment of the present invention;
FIG. 2 is a schematic diagram of a microfluidic system and a culture system according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of an internal tank of a microfluidic system according to a first embodiment of the present invention;
FIG. 4 is a schematic view showing the external structure of a culture chamber according to the first embodiment of the invention;
FIG. 5 is a top view showing the internal structure of a culture chamber according to the first embodiment of the invention;
FIG. 6 is a side view showing the internal structure of a culture chamber according to the first embodiment of the invention;
FIG. 7 is a schematic view showing a culture chamber inside a culture chamber according to a first embodiment of the invention;
FIG. 8 is a schematic diagram of a sensing system according to a first embodiment of the present invention;
FIG. 9 is a plan view showing the internal structure of a culture chamber in accordance with the second embodiment of the present invention.
Wherein: 1-microfluidic system, 2-culture tank, 3-culture chamber, 4-sealed housing, 5-liquid pump, 6-sensing system, 7-imaging analyzer, 8-computer, 9-sensing zone, 10-inner tank, 11-outer tank, 13-inlet tube, 14-inlet slot, 15-waste tube, 16-waste slot, 17-first liquid well, 18-second liquid well, 19-liquid well clamping slot, 21-first microporous filter membrane, 23-inoculation well, 24-knob, 25-aspirator, 26-sensing channel, 27-detection chamber, 28-DO probe, 29-pH probe, 30-glucose probe, 31-lactic acid probe, 32-nitric oxide probe, 33-clear water tube, 34-water tank, 35-waste tube, 36-liquid cavity, 37-culture well, 38-input channel, 39-output channel, 40-second microporous filter membrane, 41-culture plane, 42-first multiplexer, 43-second multiplexer, 44-culture cavity, 45-liquid channel, 45-liquid sensing chip, 46-microchip.
Detailed Description
In order to enhance the understanding of the present invention, the present invention will be further described in detail with reference to the drawings, which are provided for the purpose of illustrating the present invention only and are not to be construed as limiting the scope of the present invention.
Example 1
Fig. 1-8 show an automated organoid culture monitoring platform, which consists of a microfluidic system 1, a culture system, a high content system and a sensing system 6, and performs real-time monitoring of the organoid growth state through image acquisition and processing and sensors.
As shown in fig. 2 and 3, the microfluidic system 1 comprises a sealed housing 4, a liquid pump 5, a liquid inlet pipe 13 and a liquid waste pipe 15, wherein the sealed housing 4 comprises an inner tank 10 and an outer tank 11, the inner tank 10 is used for storing culture medium, the outer tank 11 is used for storing liquid waste, and a plurality of liquid cavities 36 are arranged in the inner tank 10; the top of the sealed shell 4 is provided with a liquid inlet slot 14 and a waste liquid slot 16, one end of a liquid inlet pipe 13 is connected with the culture system through a first liquid hole 17, the other end of the liquid inlet pipe is connected with a liquid pump 5 arranged in the liquid inlet slot 14, the liquid pump 5 is connected with each liquid cavity 36, the liquid pump 5 is of a multi-channel structure, culture liquid can be drawn from different liquid cavities 36, and simultaneous circulation of multiple liquids or gases in the same culture chamber 3 can be realized; one end of the waste liquid pipe 15 is connected with the outer tank 11 through a waste liquid slot 16, and the other end is connected with the culture system through a second liquid hole 18; the waste liquid pipe 15 is provided with a sensing area 9, and liquid can enter and exit the sensing system 6 through the sensing area 9. The liquid pump 5 is internally provided with a control system, different pressures can be applied by the liquid pump 5, and the culture medium in the inner tank 10 is controlled to flow into the culture system at different flow rates, so that the automatic update of the culture medium is realized, and the liquid pump 5 is automatically closed when the liquid in the inner tank 10 flows.
As shown in fig. 4 to 6, the culture system includes a plurality of culture chambers 3 provided in the culture tank 2, and culture holes 37 provided in the culture chambers 3. An environmental control device is mounted in the culture chamber 3, and the temperature, humidity, oxygen and carbon dioxide concentration in the culture chamber 3 can be controlled. The culture tank 2 is provided with codes, and each culture chamber 3 is correspondingly arranged. The culture chamber 3 is of an independent structure and is made of optically transparent original polystyrene, an inoculation hole 23 and an inoculation hole knob 24 are arranged on the upper wall, and the inoculation hole 23 can be opened by rotating the knob 24 so as to inoculate organoids into a culture hole 37 in the culture chamber 3 through the inoculation hole 23; the side wall of the culture chamber 3 is provided with a first liquid hole 17 and a second liquid hole 18, and the first liquid hole 17 and the second liquid hole 18 are provided with a first microporous filter membrane 21 with the diameter of 0.22 mu m, so that cells can be prevented from passing through. Through the liquid hole clamping groove 19 arranged outside the first liquid hole 17 and the second liquid hole 18, the liquid inlet pipe 13 and the liquid waste pipe 15 can be tightly connected with the first liquid hole 17 and the second liquid hole 18. Through the microfluidic system 1, each culture cell 3 can be simultaneously subjected to sample injection and sample discharge.
As shown in fig. 8, the sensing system 6 comprises a liquid suction device 25, a detection chamber 27, various sensing probes, a water tank 34 and the like, wherein the liquid suction device 25 sucks a fixed amount of liquid from the liquid waste pipes 15 through the sensing areas 9 connected with the liquid waste pipes 15, the liquid to be detected flows through the sensing channels 26 to enter the detection chamber 27, and the detection chamber 27 is closed after the liquid to be detected flows into the corresponding detection chamber 27; the detection chamber 27 is equipped with a plurality of sensor probes, and is capable of detecting DO, pH, glucose, and cell metabolites such as lactic acid, nitrides, and nitric oxide in the culture medium. Wherein the DO probe 28 tip consists of an optical fiber of less than 50 μm in size, a photosensitizing dye incorporated into the polymer and covering the probe 28 tip, and measuring DO concentration using dynamic luminescence quenching of the photosensitizing dye in the presence of oxygen based on the Stern-Volmer equation; the pH probe 29 comprises an iridium oxide electrode, a glass film as an interface between the liquid to be measured and the film-sealed internal buffer solution, and the pH is quantified by using the potential difference; glucose probe 30 is a platinum electrode and incorporates glucose oxidase, bovine serum albumin and glutaraldehyde cross-linking agent to track glucose levels in real time; the lactic acid probe 31 contains a platinum electrode, an immobilized lactic acid dehydrogenase, and a coenzyme (NAD or NADP) as a mediator, and electrons are shuttled between the lactic acid dehydrogenase and the electrode, and the lactic acid concentration in the culture solution is quantified by an oxidation current; the nitric oxide probe 32 contains a platinum electrode and a poly eugenol film that selectively transmits nitric oxide, and the concentration of nitric oxide is measured electrochemically. The clear water pipe 33 is connected with the sensing channel 26 and the water tank 34, deionized water in the water tank 34 is used for automatically cleaning the sensing channel 26, the detection chamber 27 and each sensing probe after each detection by the clear water pipe 33, and the liquid to be detected and cleaning waste liquid are collected into the waste liquid pipe 15 by the sewage pipe 35 after the detection is completed; the sensing system 6 is connected to a computer 8.
The high content system contains a high content cell imaging analyzer 7, and the imaging analyzer 7 can acquire, store and process organoid image data without damaging the culture environment. The culture chamber 3 accords with the specification of the high content imaging plate; the imaging assembly plate moves on an x-y axis to generate a bright field picture of each culture cell 3, an image analysis algorithm is used for automatically processing the acquired image, the contrast of the image is improved, the organoid area/non-organoid area is automatically identified according to the brightness value, the organoid diameter, area, perimeter, quantity, roundness and darkness values are quantized, and meanwhile related images and data are transmitted to a computer end 8 to form a growth curve; the imaging analyzer 7 is connected to a computer 8.
Preferably, each culture hole 37 is provided with a code, each culture hole 37 is respectively connected with an input channel 38 and an output channel 39, sample injection and sample discharge can be performed simultaneously, a second microporous filter membrane 40 with the thickness of 0.22 mu m is arranged at the connecting position, cells can be prevented from passing through, and the horizontal positions of the input channel 38 and the output channel 39 are higher than the culture plane 41; the inlet channel 38 merges into the first liquid aperture 17 at one end of the culture chamber 3 and the outlet channel 39 merges into the second liquid aperture 18 at the other end of the culture chamber 3.
Preferably, the culture hole 37 in the culture chamber 3 is made of PDMS material with good biocompatibility and has high permeability to CO 2 and O 2, so that the rapid exchange of gas between the atmosphere around the culture chamber 3 and the culture medium in the culture hole 37 is ensured; as shown in FIG. 7, a plurality of culture wells 37 are connected to each other through a liquid channel to form culture chambers 44, and each culture chamber 44 can be fed with a sample and discharged with a sample at the same time.
Preferably, a micro-sensor chip 47 is embedded in the hydrogel 46 at the bottom of the culture well 37 to obtain various parameters such as DO and pH, while monitoring nutrient and waste levels during medium renewal.
Preferably, the inner tank 10 contains a drug-containing culture medium with a fixed concentration, so that organoid high-flux drug sensitivity detection can be realized.
In the normal culture state, the inoculating hole knob 24 is opened, and the organoid is inoculated into the culture chamber 3 through the inoculating hole 23. The culture chamber 3 is connected with the inner tank 10 and the outer tank 11 through the liquid inlet pipe 13 and the waste liquid pipe 15 respectively, and is cultured while keeping proper temperature, humidity and carbon dioxide concentration, and the sensing area 9 on the waste liquid pipe 15 is connected with the liquid suction device 25 of the side wall sensing system 6 of the culture system.
Sufficient culture medium is supplemented into each liquid cavity 36 in the inner tank 10, the liquid pump 5 is turned on, and automatic liquid feeding is performed after 20min is set, then the liquid pump 5 sucks a fixed amount of culture medium after 20min, the culture medium enters each culture cell 3 of the culture system through the liquid feeding pipe 13, the culture medium is automatically stopped after the culture medium in each culture cell 3 is sufficient, the culture medium is automatically started and continuously operated after 6 hours, the flow rate is 9% of the culture cell volume of the culture medium updated every hour, and about 2mL of the culture medium is consumed every day. The waste liquid flows into the outer tank 11 through the waste liquid pipe 15.
The culture system is placed on an imaging plate of the high content cell imaging analyzer 7, the high content cell imaging analyzer 7 automatically scans and shoots, bright field pictures and organ diameter, area, perimeter, quantity, eccentricity and darkness value data are generated and transmitted to the computer 8, and when the quantity, diameter, area and perimeter of organoids are not increased any more, the roundness is reduced, and the darkness value is increased, passage prompting signals are displayed.
The liquid absorber 25 sucks a fixed amount of liquid from the culture chamber 3 through the waste liquid pipe 15, and enters the detection chamber 27 through the sensing channel 26, and the contents of DO, pH, glucose, lactic acid, nitride, nitric oxide and the like in the culture solution are detected by adopting an ion selective electrode potentiometry, amperometry and an enzyme reaction dependent biosensor in different detection chambers 27, so that continuous monitoring of the organoid culture environment is completed. When DO is reduced, pH is too small, glucose concentration is reduced, and lactic acid or nitride and nitric oxide concentration are obviously increased, the flow rate of the liquid pump 5 is automatically increased, and the whole culture chamber 3 is replaced.
Example two
As shown in fig. 9, the first and second wells are connected to the culture wells after being branched by the first and second multiplexers, respectively, and the feed tube 13 delivers the liquid to the first multiplexer 42, which in turn distributes the liquid to any one of the individual culture wells 37 of the culture chamber 3, and the output side is provided with one and the same second multiplexer 43 for guiding the liquid from each culture well 37 to the outer tank 11, allowing a very accurate dose of culture medium to be injected into each culture well 37. When one of the culture wells 37 is being treated, the state valve of the first multiplexer 42 is switched to prevent the outflow liquid from reaching any other culture well 37 by back-diffusion and back-flow, the flow rate being controlled by the number of drive cycles applied to the liquid pump 5, and the flow rate being controlled by the frequency at which the valve of the first multiplexer 42 is switched. The other structure is the same as that of the first embodiment.
The foregoing detailed description will set forth only for the purposes of illustrating the general principles and features of the invention, and is not meant to limit the scope of the invention in any way, but rather should be construed in view of the appended claims.
Claims (10)
1. An automatic change organoid culture monitoring platform, its characterized in that: comprises a micro-fluidic system (1), a culture system, a sensing system (6) and a high content system;
The micro-fluidic system (1) comprises a sealed shell (4), an inner groove (10) for containing culture solution, an outer groove (11) for containing waste liquid, a liquid pump (5), a liquid inlet pipe (13) and a waste liquid pipe (15), wherein the inner groove (10) and the outer groove (11) are arranged in the sealed shell (4), one end of the liquid inlet pipe (13) is connected with the inner groove (10) through a liquid inlet groove hole (14), the other end of the liquid inlet pipe is connected with a culture system, one end of the waste liquid pipe (15) is connected with the outer groove (11) through a waste liquid groove hole (16), the other end of the liquid pipe is connected with the culture system, the liquid pump (5) is arranged in the liquid inlet groove hole (14), and a sensing area (9) is arranged on the waste liquid pipe (15);
the culture system comprises a plurality of culture cells (3) arranged in a culture tank (2), culture holes (37) arranged in the culture cells (3) and a first microporous filter membrane (21), wherein a first liquid hole (17) and a second liquid hole (18) are formed in the side face of the culture cells (3), the liquid inlet pipe (13) and the liquid outlet pipe (15) are connected with the culture holes (37) through the first liquid hole (17) and the second liquid hole (18) respectively, an inoculation hole (23) opened and closed through a knob (24) is formed in the top of the culture cells (3) and used for inoculating organoids into the culture holes (37), and the first microporous filter membrane (21) is arranged on the first liquid hole (17) and the second liquid hole (18) and used for preventing cells from passing through;
The sensing system (6) comprises a liquid suction device (25), a detection chamber (27), a sensing probe and a water tank (34), wherein one end of the liquid suction device (25) is connected with the sensing area (9), the other end of the liquid suction device is connected with the detection chamber (27) through a sensing channel (26), the sensing probe is arranged in the detection chamber (27) and is used for detecting various parameters of culture liquid, and the water tank (34) is arranged at the bottom of the detection chamber (27) and is used for flushing the detection chamber (27);
The high content system comprises an imaging analyzer (7) and a computer (8), wherein the imaging analyzer (7) is arranged on the outer side of the culture system and used for acquiring organoid image data in a culture hole (37), and the computer (8) is connected with the imaging analyzer (7) and the sensing system (6).
2. An automated organoid culture monitoring platform according to claim 1, wherein: the inner tank (10) is internally provided with a plurality of liquid chambers (36), the liquid pump (5) is connected with each liquid chamber (36), and the liquid pump (5) is of a multi-channel structure and can draw culture liquid from different liquid chambers (36).
3. An automated organoid culture monitoring platform according to claim 1, wherein: the first liquid hole (17) and the second liquid hole (18) are respectively connected with the culture hole (37) through an input channel (38) and an output channel (39), a second microporous filter membrane (40) with the specification of 0.22 mu m is arranged at the joint, and the horizontal positions of the input channel (38) and the output channel (39) are higher than a culture plane (41) at the inner bottom of the culture hole (37).
4. An automated organoid culture monitoring platform according to claim 1, wherein: the first liquid hole (17) and the second liquid hole (18) are respectively divided into a plurality of branches by a first multiplexer (42) and a second multiplexer (43) and then are connected with the culture hole (37).
5. An automated organoid culture monitoring platform according to claim 3 or 4, wherein: the plurality of culture holes (37) are connected with each other through a liquid channel (45) to form a culture cavity (44).
6. An automated organoid culture monitoring platform according to claim 1, wherein: a micro sensing chip (47) is embedded in hydrogel (46) at the bottom of the culture hole (37) and is used for monitoring the nutrition level and the waste level in the process of medium updating.
7. An automated organoid culture monitoring platform according to claim 1, wherein: the culture chamber (3) is made of an optically transparent raw polystyrene material, and the culture hole (37) is made of a PDMS material with good biocompatibility.
8. An automated organoid culture monitoring platform according to claim 1, wherein: an environmental control device is arranged in the culture chamber (3) and can control the temperature, humidity, oxygen and carbon dioxide concentration in the culture chamber (3).
9. An automated organoid culture monitoring platform according to claim 1, wherein: the first liquid hole (17) and the second liquid hole (18) are respectively and tightly connected with the liquid inlet pipe (13) and the waste liquid pipe (15) through the liquid hole clamping groove (19).
10. An automated organoid culture monitoring platform according to claim 1, wherein: the sensing probe comprises a DO probe (28), a pH probe (29), a glucose probe (30), a lactic acid probe (31) and a nitric oxide probe (32), wherein the tip of the DO probe (28) consists of an optical fiber with the size smaller than 50 mu m, a photosensitive dye is doped into a polymer and covers the tip of the DO probe (28), and the concentration of a culture solution DO is detected by utilizing dynamic luminescence quenching of the photosensitive dye in the presence of oxygen based on a Stern-Volmer equation; the pH probe (29) comprises an iridium oxide electrode, a glass film is used as an interface between a liquid to be detected and a film sealing internal buffer solution, and the pH of the culture solution is quantified by utilizing potential difference; the glucose probe (30) is a platinum electrode, glucose oxidase, bovine serum albumin and glutaraldehyde crosslinking agent are added, and the glucose level of the culture solution is tracked in real time; the lactic acid probe (31) head contains a platinum electrode, immobilized lactic acid dehydrogenase and coenzyme as mediums to shuttle electrons between the lactic acid dehydrogenase and the electrode, and the lactic acid concentration in the culture solution is quantified through oxidation current; the nitric oxide probe (32) comprises a platinum electrode and a poly eugenol film which can selectively permeate nitric oxide, and the nitric oxide concentration of the culture solution is measured by an electrochemical method.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN118879481A (en) * | 2024-08-05 | 2024-11-01 | 长春艺夫生物科技有限公司 | Device and method for drug sensitivity testing of human intestinal cancer organoids |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118879481A (en) * | 2024-08-05 | 2024-11-01 | 长春艺夫生物科技有限公司 | Device and method for drug sensitivity testing of human intestinal cancer organoids |
| CN118879481B (en) * | 2024-08-05 | 2025-04-08 | 长春艺夫生物科技有限公司 | Equipment and method for testing drug sensitivity of human intestinal cancer organoids |
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