High-throughput microalgae cultivation device and monitoring integrated system and cultivation monitoring integrated system
Patent Information
- Application Number
- CN202522587635.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-05
- Publication Date
- 2026-10-27
- Estimated Expiration
- 2035-12-05
AI Technical Summary
然而，该方法存在固有瓶颈：（a）由于微孔板每个孔体积太小，蒸发量大，温度难以控制；（b）相邻孔之间存在严重光串扰；（c）只能采用多功能酶标仪测量OD值，误差大，成本高；（d）培养液太少导致无法做后续的成分分析；（e）使用酶标仪测量时，需要将孔板从培养箱中取出，这个过程会中断培养（温度、光照变化等），属于一种扰动式测量
[0029]1.高通量微藻培养装置通过“暗场格栅+第一匀光板+第二匀光板”的组合设计，有效消除了边缘光衰和不同培养容器间的光串扰，光照均匀度可达96%以上，为高通量平行实验提供了高度一致的培养条件。
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Figure CN224901280U_ABST
Abstract
Description
Technical Field
[0001] This utility model belongs to the field of microbial technology, specifically relating to a high-throughput microalgae culture device and integrated monitoring system, as well as a culture and monitoring integrated system. Background Technology
[0002] Optimizing microalgae culture conditions (such as nutrient ratios) requires numerous parallel experiments. Traditional methods mainly involve two technical approaches, but both have significant drawbacks.
[0003] The glass Erlenmeyer flask culture method is the most basic method, but it is cumbersome, time-consuming, labor-intensive, consumes a lot of culture medium, and is difficult to achieve uniformity of light among a large number of parallel samples, and cannot be used for high-throughput screening.
[0004] Microplate culture methods utilize 24-well or 96-well microplates to increase throughput. However, this method has inherent bottlenecks: (a) due to the small volume of each well, evaporation is high, making temperature control difficult; (b) severe optical crosstalk exists between adjacent wells; (c) only multi-mode microplate readers can be used to measure OD values, resulting in large errors and high costs; (d) insufficient culture medium prevents subsequent component analysis; and (e) when using a microplate reader, the well plate needs to be removed from the incubator, interrupting the culture process (due to temperature, light changes, etc.), making it a perturbation-based measurement.
[0005] In summary, neither the traditional triangular flask method nor the microplate method can achieve non-destructive, in-situ, and real-time dynamic monitoring of each parallel sample during the culture process. Utility Model Content
[0006] One of the objectives of this invention is to provide a high-throughput microalgae culture device. The composite structure of "first light-uniform plate, second light-uniform plate and dark field grid" enables each culture container to have high light uniformity, meeting the requirements of high-throughput culture. The top cover gap design effectively reduces the evaporation rate and ensures long-term culture stability.
[0007] Another objective of this invention is to provide an integrated high-throughput microalgae culture and monitoring system that includes the above-mentioned high-throughput microalgae culture device, which can perform in-situ, non-destructive monitoring of the cultured microalgae while achieving high-throughput culture.
[0008] The technical solution of this utility model is as follows:
[0009] A high-throughput microalgae cultivation device includes a cultivation module and a light module;
[0010] The training module includes:
[0011] Dark field grille, with opaque sidewalls and an array of through-holes inside;
[0012] The first light-diffusing plate is fixed to the bottom of the dark field grid, and the dark field grid and the first light-diffusing plate together form an array of receiving cavities.
[0013] A culture container with a transparent bottom is disposed within a receiving cavity, and the shape and size of the culture container are adapted to the shape and size of the receiving cavity;
[0014] The top cover is transparent and covers the dark field grid, and a gas exchange gap is provided between the top cover and the upper surface of the dark field grid.
[0015] The illumination module is located below the culture module and includes a culture light source and a second light homogenizing plate. The second light homogenizing plate is located between the first light homogenizing plate and the culture light source, and the projected area of the second light homogenizing plate is larger than the projected area of the dark field grid.
[0016] In some preferred embodiments, the culture container is a cuvette.
[0017] In some preferred embodiments, the projected area of the second light-diffusing plate is at least 1.2 times the projected area of the dark field grid.
[0018] In some preferred embodiments, the gap between the top cover and the upper surface of the dark field grid is 1 mm.
[0019] In some preferred embodiments, the cultivation light source provides the spectrum required for photosynthesis in microalgae and can adjust the light intensity and light-dark cycle according to cultivation needs.
[0020] A high-throughput microalgae culture monitoring integrated system includes the above-mentioned high-throughput microalgae culture device, a measurement light source, an in-situ optical monitoring module, and a control and data processing unit;
[0021] The measurement light source is positioned below the culture module;
[0022] The in-situ optical monitoring module includes an XY-axis moving frame and an optical measuring head; the XY-axis moving frame is mounted on top of the top cover; the optical measuring head is mounted on the XY-axis moving frame and is configured to perform non-contact optical monitoring of the culture container through the top cover.
[0023] The control and data processing unit is electrically connected to the XY-axis moving frame, the optical measuring head, the measuring light source, and the culture light source. The control and data processing unit is configured to control the XY-axis moving frame to move the optical measuring head above the culture container to be measured to acquire optical signals and process the acquired optical signals.
[0024] In some preferred embodiments, the high-throughput microalgae culture and monitoring integrated system further includes a temperature-controlled shaker, in which the culture module, illumination module, and in-situ optical monitoring module are placed, and the temperature-controlled shaker is connected to the control and data processing unit.
[0025] In some preferred embodiments, the measuring light source is a halogen lamp or an LED light source.
[0026] In some preferred embodiments, the emission spectrum of the measuring light source covers the characteristic wavelengths of the analyte.
[0027] In some preferred embodiments, the optical measuring head is a miniature spectrometer or a CMOS camera.
[0028] This utility model has at least the following beneficial effects:
[0029] 1. The high-throughput microalgae culture device effectively eliminates edge light decay and light crosstalk between different culture containers through the combined design of "dark field grid + first light uniform plate + second light uniform plate". The light uniformity can reach more than 96%, providing highly consistent culture conditions for high-throughput parallel experiments.
[0030] 2. The tiny gap (e.g., 1 mm) between the top cover of the high-throughput microalgae culture device and the upper surface of the dark field grid ensures gas exchange while significantly reducing the evaporation rate of the culture medium (as low as 1.8% in the test), thus ensuring the stability of long-term culture.
[0031] 3. In some preferred embodiments, conventional culture containers such as standard cuvettes are used, which can be directly transferred to a common spectrophotometer for measurement, reducing the dependence on expensive special equipment (such as ELISA readers), saving equipment costs and avoiding sample transfer contamination.
[0032] 4. The array-type containment chamber design consisting of a dark field grid and a first light homogenizing plate supports dozens to hundreds of parallel experiments. The amount of culture medium required for a single sample is small, which greatly saves algae and reagents and achieves high-throughput culture with minimal space occupation.
[0033] 5. The high-throughput microalgae culture monitoring integrated system uses an optical measuring head mounted on an XY-axis moving frame to perform non-contact optical scanning of the culture container through a transparent top cover, enabling real-time, non-destructive dynamic monitoring without interrupting the culture process. Attached Figure Description
[0034] Figure 1 This is a schematic diagram of the high-throughput microalgae cultivation device provided in Example 1, wherein the top cover is not placed above the dark field grid;
[0035] Figure 2 A side view of the high-throughput microalgae culture device provided in Example 1;
[0036] Figure 3 This is a schematic diagram of the dark field grid structure in Example 1;
[0037] Figure 4 A side view of the high-throughput microalgae culture monitoring integrated system provided in Example 1;
[0038] Figure 5 This is a schematic diagram of the integrated high-throughput microalgae culture and monitoring system provided in Example 1;
[0039] Figure 6 The flowchart shows the workflow of the integrated high-throughput microalgae culture and monitoring system provided in Example 1.
[0040] The reference numerals in the figure are: 1-Dark field grid; 101-Through hole structure; 2-First light homogenizer; 3-Second light homogenizer; 4-Top cover; 5-Cultivation light source; 6-Optical measuring head; 7-XY axis moving frame. Detailed Implementation
[0041] The technical solution of this utility model will be further explained and described below through specific embodiments.
[0042] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are preferred embodiments of the present utility model and should not be considered as excluding other embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present utility model without creative effort are within the scope of protection of the present utility model.
[0043] Unless otherwise expressly defined, in the claims, description and accompanying drawings of this utility model, the use of directional terms such as "top", "bottom", "inner", "outer", "upper", "lower", "front", "rear" to indicate orientation or positional relationship is based on the orientation and positional relationship shown in the accompanying drawings, and is only for the convenience of describing this utility model and simplifying the description, and is not intended to indicate or imply that the device or element referred to must have a specific orientation or be constructed and operated in a specific orientation, and therefore should not be construed as limiting the specific protection scope of this utility model.
[0044] The orthographic projection mentioned in this article refers to the projection from the first homogenizing plate to the second homogenizing plate.
[0045] In Example 1, the projected area of the first light-diffusing plate is slightly larger than the projected area of the dark field grid. The phrase "slightly larger" is used because the first light-diffusing plate also acts as a base; after the top cover is placed on the dark field grid, the lower edge of the top cover abuts against the upper surface of the first light-diffusing plate. Therefore, those skilled in the art should understand that the area of the first light-diffusing plate matches the shape of the lower edge of the top cover, and that the phrase "slightly larger" does not make the embodiment unclear. Of course, in other possible embodiments, the projected area of the first light-diffusing plate only needs to be not smaller than the projected area of the dark field grid, and the first light-diffusing plate can close the bottom of the dark field grid.
[0046] Furthermore, the first light-diffusing plate is fixedly connected to the bottom of the dark-field grid, thereby sealing the bottom of the through-hole structure. Therefore, the sidewalls of the dark-field grid and the upper surface of the first light-diffusing plate together form an array-type containment cavity. It can be understood that the containment cavity is functionally a bottom-transparent cavity used to stably hold the culture container.
[0047] Example 1
[0048] This embodiment first provides a high-throughput microalgae cultivation device, including, as follows: Figure 1 and Figure 2 The cultivation module and illumination module are shown.
[0049] Specifically, the culture module consists of a dark field grid 1, a first light-diffusing plate 2, a culture container (not shown in the figure), and a top cover 4.
[0050] Among them, such as Figure 3 The dark field grid 1 shown is divided into several arrays of through-hole structures 101 that are arranged vertically. The dark field grid 1 is made of black opaque ABS (acrylonitrile-butadiene-styrene terpolymer) by 3D printing. The shape and size of its through-hole structure 101 are adapted to the culture container.
[0051] Back Figure 1 and Figure 2 The first light-diffusing plate 2 is fixed to the bottom of the dark field grid 1 and seals the bottom of the dark field grid 1. The first light-diffusing plate 2 and the dark field grid 1 together form a plurality of arrayed receiving cavities. The side walls of the receiving cavities are made of black opaque material, which can effectively prevent light crosstalk between different receiving cavities. With the first light-diffusing plate 2 as the bottom, the uniformity of the culture light / monitoring light received by different receiving cavities can be improved.
[0052] The culture container, having a transparent bottom and a shape and size adapted to the culture chamber, is placed within the chamber. In this embodiment, the culture container is a standard cuvette. In other possible embodiments, the culture container can be other micro-containers with transparent bottoms. Compared to the traditional Erlenmeyer flask culture method, the volume of culture medium required for a single culture container is reduced by 1 to 2 orders of magnitude. When conducting dozens to hundreds of parallel experiments, this significantly saves on precious algal stockpiles, expensive culture medium additives, and chemical reagents, while minimizing space occupation. Compared to the ordinary microplate culture method, this embodiment allows the culture container to be directly transferred to a standard spectrophotometer for endpoint comparison measurements, completely eliminating the reliance on expensive specialized equipment such as multifunctional microplate detectors. This greatly reduces the equipment threshold and operating costs, and avoids the risk of contamination from sample transfer.
[0053] The top cover 4 is made of transparent glass and is positioned above the dark field grid 1, forming a 1mm gap between it and the upper surface of the dark field grid 1. This balances the conflict between carbon dioxide supply and water evaporation loss in microalgae cultivation, ensuring the stability of long-term cultivation. In other possible embodiments, the top cover 4 can also be made of other colorless and transparent materials, such as colorless and transparent acrylic.
[0054] The illumination module includes a culture light source 5 and a second light homogenizing plate 3.
[0055] The cultivation light source 5 is positioned below the cultivation module and emits cultivation light upwards. The cultivation light source 5 is configured to provide the spectrum required for microalgal photosynthesis (such as blue light, red light, or full spectrum) and can adjust the light intensity and light-dark cycle according to cultivation needs.
[0056] The second homogenizing plate 3 is disposed between the culture light source 5 and the culture module. In this embodiment, the first homogenizing plate 2, the second homogenizing plate 3, and the dark field grid 1 are all cubes, and their geometric centers are located on the same vertical axis. The projected area of the first homogenizing plate 2 is slightly larger than that of the dark field grid 1, and the projected area of the second homogenizing plate 3 is 1.44 times that of the first homogenizing plate 2. In other possible embodiments, the projected area of the second homogenizing plate 3 is not less than 1.2 times that of the first homogenizing plate 2. This application can effectively compensate for edge light attenuation through the first homogenizing plate 2 and the second homogenizing plate 3. Tests have shown that, after the optical isolation of the dark field grid 1 and the coordinated homogenization of the first homogenizing plate 2 and the second homogenizing plate 3, the illumination uniformity within the device can reach over 95%, providing highly consistent culture conditions for high-throughput screening.
[0057] like Figure 4As shown in the figure, this embodiment also provides a high-throughput microalgae culture monitoring integrated system, including the above-mentioned high-throughput microalgae culture device, measurement light source, in-situ optical monitoring module, temperature-controlled shaker (not shown in the figure) and control and data processing unit (not shown in the figure).
[0058] The measuring light source and the culture light source 5 are integrated in the same lamp box and can be controlled independently. The measuring light source is a halogen lamp, which has a high color rendering index and its emission spectrum covers the characteristic wavelength of the analyte. In other possible embodiments, the measuring light source may also be monochromatic light of the characteristic wavelength of the analyte.
[0059] The in-situ optical monitoring module includes an XY-axis moving frame 7 and an optical measuring head 6. The XY-axis moving frame 7 is mounted above the top cover 4 and can be precisely positioned in the X and Y-axis planes under the drive of a motor. The optical measuring head 6 is mounted on the XY-axis moving frame 7 and configured to perform non-contact optical monitoring of the culture container through the top cover 4. In this embodiment, the optical measuring head 6 is a miniature spectrometer, capable of acquiring full-spectrum data from the culture container, achieving real-time, non-destructive, high-throughput monitoring, facilitating subsequent analysis of multiple key physiological parameters such as biomass concentration, pigment composition and content, and physiological state. In other possible embodiments, the optical measuring head 6 is a common CMOS camera, enabling low-cost, high-speed monitoring of biomass.
[0060] The aforementioned culture module, illumination module, measurement light source, and in-situ optical monitoring module are housed in a temperature-controlled shaker. The temperature-controlled shaker provides a closed, stable, and controllable temperature and humidity environment, ensuring that each culture chamber is under identical temperature conditions and achieving stable evaporation control. Simultaneously, the temperature-controlled shaker promotes gas exchange through oscillation, prevents microalgae sedimentation and adhesion to the walls, and promotes uniform nutrient distribution within the culture chamber, thereby ensuring the reliability and reproducibility of high-throughput screening data.
[0061] The control and data processing unit is electrically connected to the XY-axis moving frame 7, the optical measuring head 6, the measuring light source, and the culture light source 5. The control and data processing unit is configured to control the XY-axis moving frame to move the optical measuring head above the culture container to be measured to acquire and process the acquired optical signals.
[0062] The following describes the workflow of the integrated high-throughput microalgae culture and monitoring system provided in this embodiment. Figure 5 and Figure 6 As shown, it includes the following operations: culture, in-situ monitoring (triggering, light source switching, scanning and acquisition, and looping), and data processing.
[0063] (1) Trigger: The control and data processing unit triggers the monitoring cycle according to the preset time (e.g., every 1 hour) or external signal.
[0064] (2) Light source switching: The control and data processing unit first stops the operation of the temperature-controlled shaker, restores the temperature-controlled shaker to its initial position, and then turns off the culture light source 5. After waiting for a short delay (e.g., 500 milliseconds to ensure that the culture light source 5 is completely extinguished), the measurement light source is turned on. At this time, only the measurement light source is working below the device, providing pure and stable illumination for optical detection.
[0065] (3) Scanning and Acquisition: The XY-axis moving frame 7 moves the optical measuring head 6 above the first culture container to be measured (e.g., position A1). After the XY-axis moving frame 7 stabilizes (e.g., pause for 0.5 seconds to eliminate vibration), the control and data processing unit instructs the optical measuring head 6 to perform integration and acquisition to obtain the transmission spectrum (optical signal) of that point. The measurement cycle of a single point can be controlled within 2 seconds.
[0066] (4) Cycle: Repeat step 3 until all culture containers to be measured (e.g., 10x10 array, 100 points in total) have been scanned. It takes about 3.5 minutes to complete the entire scan.
[0067] (5) Cultivation operation: After scanning all points, the control and data processing unit turns off the measurement light source and immediately turns on the cultivation light source 5 and the temperature-controlled shaker to restore the microalgae to normal photosynthetic cultivation.
[0068] (6) Data processing operation: The control and data processing unit calculates the absorbance at each characteristic wavelength based on the spectral data collected by the optical measuring head 6 and plots the time series curve.
[0069] Example 2: Comparative Experiment of Dynamic Growth Monitoring and Endpoint Screening
[0070] This embodiment aims to demonstrate the comprehensive advantages of the present invention over traditional methods in terms of high-throughput screening efficiency, data quality, and downstream application scalability.
[0071] Objective: To verify whether the high-throughput microalgae culture monitoring integrated system provided by this invention can identify high-performance algae strains earlier and more accurately than the traditional endpoint method by dynamically monitoring growth kinetic parameters (dynamic method), and to evaluate its ability to perform subsequent in-depth analysis.
[0072] Experimental Design:
[0073] One known fast-growing wild-type algal strain (WT) and five mutant algal strains (MT1-MT5) were selected. All algal strains were cultured in parallel under the same conditions (n=5) in the high-throughput microalgae culture monitoring system, Erlenmeyer flasks, and 96-well plates provided in Example 1.
[0074] The high-throughput microalgae culture monitoring system automatically scans all culture containers every 2 hours to monitor the OD750 value for 7 consecutive days. Both the Erlenmeyer flask group and the 96-well plate group were analyzed using the endpoint method. In the Erlenmeyer flask group, samples were taken on the 7th day, and the endpoint biomass was calculated by measuring the OD750 value (traditional endpoint method). In the 96-well plate group, the endpoint biomass was calculated by measuring the OD750 value on the 7th day using a microplate reader (existing high-throughput endpoint method).
[0075] Data Analysis and Downstream Applications: The dynamic method uses data from the first 72 hours to fit the growth curve and calculate the maximum specific growth rate (μmax). After cultivation, an appropriate amount of algal solution is taken directly from the culture container (cuvette) for lipid content determination. Due to insufficient culture medium volume, the 96-well plate group could not perform component analysis under the same batch and conditions.
[0076] Results and comparisons:
[0077] Screening timeliness: The dynamic method can complete the screening based on μmax on the 3rd day, while the endpoint method (Eragonal flask group and 96-well plate group) must wait until the 7th day, shortening the screening cycle by 57%.
[0078] Screening accuracy: The endpoint method showed that MT1 had the highest final biomass. However, the dynamic method revealed that WT's μmax was significantly higher than MT1 (0.08 h⁻¹ vs. 0.05 h⁻¹), indicating that WT had superior growth potential. Subsequent validation experiments confirmed that in fed-batch culture, WT's biomass yield surpassed MT1 by 20%.
[0079] Downstream analytical advantages: Dynamic analysis of the algal broth from the WT strain revealed a lipid content of 35% of the cell dry weight. This successful acquisition of this key physiological indicator directly correlates growth kinetic advantages with the final economic trait (oil production capacity), achieving a seamless transition from "rapid initial screening" to "precise validation." In contrast, the 96-well plate method only provides endpoint biomass data and cannot perform such subsequent component analysis, leading to uncertainty in its screening results.
[0080] Conclusion: The dynamic method not only significantly improves screening speed but also reveals potential growth advantages through dynamic parameters, avoiding misjudgments that may occur with the endpoint method due to different times when strains enter the decline phase. Its ample sample volume provides valuable downstream analysis samples for high-throughput screening, forming a complete "screening-validation" closed loop, which is impossible with the microplate method.
[0081] Example 3: Dynamic analysis of lipid accumulation in microalgae under nitrogen stress
[0082] This embodiment aims to demonstrate the unique ability of this invention to reveal subtle physiological processes, which is impossible with traditional methods.
[0083] Objective: To capture and quantify the continuous dynamic changes in physiological indicators of microalgae in response to nutrient stress, rather than simply observing the final results.
[0084] Experimental Design: An oil-producing microalga was initially cultured normally. During the mid-exponential growth phase, the culture medium was switched to nitrogen-free medium to induce lipid accumulation. The high-throughput microalgae culture monitoring system provided in Example 1 was activated, and a high-frequency full scan was performed every 30 minutes for continuous monitoring for 96 hours. Using full-spectrum data, in addition to biomass (OD750), the OD530 / OD680 ratio was analyzed as an indirect indicator of neutral lipid accumulation using a specific algorithm. After monitoring, the culture containers at specific time points were immediately removed, and the algae were analyzed for the biochemical composition of proteins, carbohydrates, and lipids.
[0085] Results and scientific findings:
[0086] Response timing: Dynamic data clearly show that the growth rate began to decline 7.5 hours after nitrogen stress, while the lipid accumulation signal significantly increased after 12.2 hours. Traditional methods cannot capture this precise physiological response timing.
[0087] Process kinetics: Nonlinear kinetic curves of lipid accumulation were successfully plotted, revealing a burst growth period rather than uniform accumulation 24-48 hours after stress.
[0088] The fundamental difference from the 96-well plate method is that precise biochemical assays can be performed directly using the same culture sample at the peak point of lipid accumulation rate changes (e.g., 48 hours after stress). This achieves a precise correspondence between "dynamic process monitoring data" and "static endpoint component data" on the same sample, providing the only feasible means to construct reliable physiological models. With 96-well plates, due to sample volume limitations, destructive sampling must be performed at the expense of parallel samples, or such analyses may be impossible to complete, leading to a disconnect between process and results.
[0089] Conclusion: This invention provides a high temporal resolution physiological dynamic map, offering crucial data for analyzing the precise time windows of metabolic pathway regulation, and achieving a leap from "outcome observation" to "process analysis." It also overcomes the sample size bottleneck that microplate culture systems cannot overcome.
[0090] Example 4: Illumination Uniformity Test
[0091] Methods and Results: Illuminance values at the centers of 100 culture chambers within the culture module of Example 1 were measured using a lux meter. The average illuminance was 2460 lux, the minimum was 2390 lux, and the maximum was 2480 lux. The calculated light uniformity (minimum / maximum) was as high as 96.4%.
[0092] Brief conclusion: The results confirm that the double-layer homogenization system consisting of the second homogenizing plate and the first homogenizing plate can provide a highly uniform culture environment, effectively eliminate edge light decay, and provide a fundamental guarantee for the reliability of each parallel experiment.
[0093] Example 5: Evaporation Control Test
[0094] Methods and Results: Under an environment of 20°C and 50% humidity, the evaporation of the culture medium in a traditional 96-well plate and the culture module (using cuvettes as the culture device) in Example 1 was compared during a 7-day static incubation period. The results showed that the average evaporation rate of the culture medium in the 96-well plate was 6.5%, while the average evaporation rate in the device of this invention was only 1.8%.
[0095] In summary, this invention effectively controls the evaporation of the culture medium and ensures the stability of long-term culture experiments by using a standard cuvette as the culture container and combining it with the uniform gap formed between the top cover and the dark field grid.
[0096] The above are merely preferred embodiments of the present utility model, and therefore cannot be used to limit the scope of the present utility model. All equivalent changes and modifications made in accordance with the scope of the present utility model patent and the contents of the specification should still fall within the scope of the present utility model.
Claims
1. A high-throughput microalgae cultivation device, characterized in that, Includes a cultivation module and a lighting module; The cultivation module includes: Dark field grille, with opaque sidewalls and an array of through-holes inside; The first light-diffusing plate is fixed to the bottom of the dark field grid, and the dark field grid and the first light-diffusing plate together form an array-arranged receiving cavity. A culture container having a transparent bottom is disposed within the receiving cavity, and the shape and size of the culture container are adapted to the shape and size of the receiving cavity; A top cover, the top surface of which is transparent and covers the dark field grid, and a gas exchange gap is provided between the top cover and the upper surface of the dark field grid. The illumination module is located below the culture module and includes a culture light source and a second light-diffusing plate; the second light-diffusing plate is located between the first light-diffusing plate and the culture light source, and the projected area of the second light-diffusing plate is larger than the projected area of the dark field grid.
2. The high-throughput microalgae cultivation device as described in claim 1, characterized in that, The culture container is a cuvette.
3. The high-throughput microalgae cultivation device as described in claim 1, characterized in that, The projected area of the second light-diffusing plate is at least 1.2 times the projected area of the dark field grid.
4. The high-throughput microalgae cultivation device as described in claim 1, characterized in that, The gap between the top cover and the upper surface of the dark field grid is 1 mm.
5. The high-throughput microalgae cultivation device as described in claim 1, characterized in that, The culture light source provides the spectrum required for photosynthesis in microalgae and can adjust the light intensity and light-dark cycle according to the culture requirements.
6. A high-throughput integrated system for monitoring microalgae culture, characterized in that, Includes the high-throughput microalgae cultivation device, measurement light source, in-situ optical monitoring module, and control and data processing unit as described in any one of claims 1 to 5; The measuring light source is positioned below the culture module; The in-situ optical monitoring module includes an XY-axis moving frame and an optical measuring head; the XY-axis moving frame is mounted above the top cover; the optical measuring head is mounted on the XY-axis moving frame and is configured to perform non-contact optical monitoring of the culture container through the top cover. The control and data processing unit is electrically connected to the XY-axis moving frame, the optical measuring head, the measuring light source, and the culture light source. The control and data processing unit is configured to control the XY-axis moving frame to move the optical measuring head above the culture container to be measured to acquire optical signals and process the acquired optical signals.
7. The integrated high-throughput microalgae culture and monitoring system as described in claim 6, characterized in that, It also includes a temperature-controlled shaker, in which the culture module, the illumination module and the in-situ optical monitoring module are placed, and the temperature-controlled shaker is connected to the control and data processing unit.
8. The integrated high-throughput microalgae culture and monitoring system as described in claim 6, characterized in that, The measurement light source is a halogen lamp or an LED light source.
9. The integrated high-throughput microalgae culture and monitoring system as described in claim 6, characterized in that, The emission spectrum of the measuring light source covers the characteristic wavelengths of the analyte.
10. The integrated high-throughput microalgae culture and monitoring system as described in claim 6, characterized in that, The optical measuring head is a miniature spectrometer or a CMOS camera.