Biological culture and imaging integrated chip and three-dimensional imaging system

Abstract

The invention provides a biological culture and imaging integrated chip and a three-dimensional observation system. The biological culture and imaging integrated chip comprises a microfluidic structure and an image acquisition part, a culture cavity is formed in the micro-fluidic structure; the culture cavity is of a porous structure; the culture cavity is used for accommodating biological samples; the image acquisition part comprises an acquisition device; the collection device is attached to an orifice of the culture cavity; the acquisition device is used for acquiring an image of the biological sample in the culture cavity; in the embodiment of the invention, the biological culture and imaging integrated chip can effectively shorten the spacing distance between the biological sample in the culture cavity and the acquisition device in the image acquisition part and the number of dielectric layers, and eliminates three-dimensional reconstruction errors caused by gaps and multiple dielectric layers in a traditional microscopic method; therefore, potential errors caused by signal attenuation or logistics model errors are remarkably reduced, and finally the accuracy and reliability of acquisition and imaging of the biological sample are improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of microbial culture detection, in particular to a biological culture imaging integrated chip and a three-dimensional imaging system. BACKGROUND

[0002] In the application of long-term organ micro-imaging, it has significant technical advantages to realize the miniaturization of imaging equipment and integrate it in the incubator for in-situ observation. At present, the existing imaging technology related to the field mainly includes traditional optical micro-imaging and lensless imaging. The traditional optical micro-imaging usually places a sample carrier such as a culture dish or a microfluidic chip on the objective table of a standard microscope. Through a complex optical system composed of an objective lens and other precise optical components, the illumination light source is focused on the sample area, and an enlarged optical image is formed on the remote photodetector. Lensless imaging is to place the culture dish or the substrate carrying the sample directly at a very close distance (usually the distance is less than 5 mm) from the photosensitive effective area of the CMOS image sensor chip. It relies on the direct modulation of light by the sample (such as diffraction, absorption or phase modulation) to form a projection image or interference pattern directly on the sensor surface, thereby eliminating the lens group in the traditional imaging system.

[0003] In the use process of the existing imaging technology, there is usually a working distance of several millimeters between the biological sample and the photosensitive surface of the image sensor. The path of the light signal propagating from the sample to the image sensor must pass through multiple medium layers with different refractive indices and thicknesses, including the sample carrier (such as a glass slide), an air layer, and the packaging protection layer of the CMOS chip itself, possibly a filter or a microlens. The inhomogeneity of the medium in this optical path will cause complex optical distortion and signal attenuation, seriously interfere with the model-based image reconstruction process, significantly reduce the fidelity and accuracy of the reconstructed image, and put higher requirements on the adaptability and calibration of the algorithm. SUMMARY

[0004] In order to solve the above technical problems, the present application discloses a biological culture imaging integrated chip and a three-dimensional imaging system, which can solve the technical problems that the inhomogeneity of the medium in the optical path will cause complex optical distortion and signal attenuation, seriously interfere with the model-based image reconstruction process, significantly reduce the fidelity and accuracy of the reconstructed image, and put higher requirements on the adaptability and calibration of the algorithm.

[0005] In order to achieve the above purpose, the present application provides a biological culture imaging integrated chip, which comprises a microfluidic structure and an image acquisition part.

[0006] The microfluidic structure is provided with a culture cavity; the culture cavity is a hole structure; and the culture cavity is used for accommodating a biological sample.

[0007] The image acquisition part comprises an acquisition device; the acquisition device is arranged at the orifice of the culture cavity; and the acquisition device is used for acquiring the image of the biological sample in the culture cavity.

[0008] In a possible embodiment, the microfluidic structure is provided with at least one flow channel; one end of the flow channel is in communication with the culture cavity, and the other end of the flow channel is in communication with the outside world.

[0009] The biological sample can flow into or out of the culture cavity through the flow channel.

[0010] In a possible embodiment, the flow channel is in communication with the top of the culture cavity.

[0011] In a possible embodiment, the culture cavity is a through-hole structure with two orifices.

[0012] The biological culture imaging integrated chip further comprises a transparent cover plate.

[0013] In the process of acquiring the image of the biological sample in the culture cavity by the acquisition device, the acquisition device is arranged at one orifice of the through-hole structure, and the transparent cover plate is arranged at the other orifice of the through-hole structure.

[0014] In a possible embodiment, the microfluidic structure is made of a material comprising polydimethylsiloxane.

[0015] The polydimethylsiloxane material is a material prepared by using a prepolymer and a crosslinking agent as raw materials, through light curing, and in a mold with low surface roughness.

[0016] In a possible embodiment, the acquisition device comprises a complementary metal oxide semiconductor image sensor, or the acquisition device comprises a charge coupled device sensor.

[0017] In another aspect, the present application discloses a three-dimensional imaging system, which comprises any one of the biological culture imaging integrated chips, at least one illumination lamp strip, and an electric control system.

[0018] Each illumination lamp strip comprises at least two illumination devices; every two adjacent illumination devices in the at least two illumination devices are spaced apart by a preset angle; and each illumination device is arranged towards the culture cavity.

[0019] In the use of the three-dimensional imaging system, each illumination device on each illumination lamp strip is sequentially turned on and turned off, so that the image acquisition part can acquire images when each illumination device illuminates the culture cavity; and the electric control system can establish a three-dimensional model of the biological sample through the acquisition results of the image acquisition part.

[0020] In a possible embodiment, the three-dimensional imaging system comprises two arc-shaped illumination lamp strips.

[0021] The image collector is located at the bottom of the biological culture and imaging integrated chip; and the two orthogonal illumination lamp strips are arranged above the biological culture and imaging integrated chip.

[0022] In a possible embodiment, the biological culture and imaging integrated chip is located at the arc center of the two arc-shaped illumination lamp strips.

[0023] In a possible embodiment, the shielding member is arranged on each illumination device.

[0024] The shielding member is used to limit the light angle of the illumination device so as to concentrate the illumination of the culture cavity.

[0025] The technical scheme provided by the embodiment of the application has the following technical effects:

[0026] 1. The biological culture and imaging integrated chip can integrate the microfluidic structure and the image collector, thereby obtaining a small and highly integrated biological culture and imaging structure, which is suitable for long-term biological sample imaging.

[0027] 2. In the prior art, the biological sample in the culture dish is observed by placing the culture dish containing the biological sample on the image sensor. In this observation process, the light signal must pass through multiple medium layers with different refractive indexes and thicknesses, including the sample carrier (the transparent bottom plate of the culture dish), the air layer and the packaging protection layer of the image sensor itself, in the path from the biological sample to the image sensor. The biological culture and imaging integrated chip directly matches the collector with the aperture of the culture cavity, so that the collector is matched with the biological sample in the culture cavity, effectively shortens the interval distance between the biological sample in the culture cavity and the collector in the image collector and the number of medium layers, eliminates the three-dimensional reconstruction error caused by the gap and the multiple medium layers in the traditional microscopic method, thereby significantly reducing the potential error introduced by signal attenuation or flow model error, and finally improving the accuracy and reliability of the biological sample imaging.

[0028] 3. The biological culture and imaging integrated chip integrates the culture and observation of the biological sample, thereby realizing the miniaturization of the chip, making the structure more compact and smaller, and being more suitable for in-box long-term observation than the prior art. BRIEF DESCRIPTION OF DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, and the advantages thereof, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description are only some embodiments of the present application, and for those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0030] Figure 1 is a schematic diagram of a biological culture imaging integrated chip provided by an embodiment of the present application;

[0031] Figure 2 is a schematic diagram of a biological culture imaging integrated chip provided by an embodiment of the present application after a transparent cover plate is covered on the cover of the biological culture imaging integrated chip;

[0032] Figure 3 is a schematic diagram of a three-dimensional imaging system provided by an embodiment of the present application;

[0033] Figure 4 is a schematic diagram of a historical modeling of a three-dimensional imaging system provided by an embodiment of the present application;

[0034] In the drawings, the reference signs are explained as follows:

[0035] 1-microfluidic structure; 11-culture cavity; 12-flow channel; 2-image acquisition unit; 3-transparent cover plate; 4-illumination lamp strip; 41-illumination device. DETAILED DESCRIPTION

[0036] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without any creative effort belong to the scope of protection of the present application.

[0037] It should be noted that the "one embodiment" or "an embodiment" described in the specification of the present application means that a specific feature, structure or characteristic described in the specification can be included in at least one implementation of the present application. It should be understood that in the specification and claims of the embodiments of the present application and the above-mentioned drawings, the terms "upper", "lower", "top", "bottom" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the present application. The terms "first", "second" are only for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features limited by "first", "second" can be explicitly or implicitly included one or more features. Moreover, the terms "first", "second" and the like are used to distinguish similar objects, and do not necessarily describe a particular order or sequence.

[0038] As shown in Figures 1 to 2 The present application provides a biological culture imaging integrated chip, which comprises a microfluidic structure 1 and an image acquisition part 2. The microfluidic structure 1 can be a structure for simulating the microenvironment in the biological body in the biological culture imaging integrated chip, and the microfluidic structure 1 can provide dynamic culture conditions for cells or tissues. The image acquisition part 2 can be a functional module or functional device capable of acquiring images of biological samples.

[0039] In a possible embodiment, the microfluidic structure 1 is provided with a culture cavity 11, which can be a hole structure. The culture cavity 11 can be a hole structure including a cylindrical hole, a square hole, an irregular hole, or any shaped cavity designed according to actual production and life needs. The culture cavity 11 is used to accommodate biological samples including biological cells, biological tissues and organoids.

[0040] Optionally, the image acquisition part 2 can include an acquisition device, which is attached to the aperture of the culture cavity 11. The acquisition device can be attached to the aperture of the culture cavity 11 and fixedly connected with the microfluidic structure 1, or the acquisition device can be attached to the aperture of the culture cavity 11 and adhesively connected with the microfluidic structure 1. The acquisition device can be used to acquire images of biological samples in the culture cavity 11.

[0041] In this embodiment, the bio-culture imaging integrated chip can integrate the microfluidic structure 1 and the image acquisition unit 2, thereby obtaining a bio-culture imaging structure with small volume and high integration, which is suitable for long-term bio-sample imaging. Meanwhile, the bio-culture imaging integrated chip can effectively shorten the distance between the bio-sample in the culture cavity 11 and the acquisition device in the image acquisition unit 2 and the number of medium layers, eliminate the three-dimensional reconstruction error caused by the gap and the multiple medium layers in the traditional microscopic method, significantly reduce the potential error caused by signal attenuation or flow model error, and finally improve the accuracy and reliability of bio-sample imaging.

[0042] In a possible embodiment, the microfluidic structure 1 is provided with at least one flow channel 12, one end of the flow channel 12 is in communication with the culture cavity 11, and the other end of the flow channel 12 is in communication with the outside, so that the bio-sample can flow into or out of the culture cavity 11 through the flow channel 12.

[0043] Optionally, the flow channels 12 in the at least one flow channel 12 can be divided into an inlet flow channel and an outlet flow channel, the bio-sample can flow into the culture cavity 11 from the inlet flow channel, and can flow out of the culture cavity 11 through the outlet flow channel, thereby realizing the flow balance of the liquid in the microfluidic structure 1.

[0044] In a possible embodiment, the flow channel 12 is in communication with the top of the culture cavity 11. The top of the flow channel 12 can be flush with the top of the culture cavity 11, and the depth of the flow channel 12 can be less than the depth of the culture cavity 11, so that the bio-sample can flow into or out of the culture cavity 11 through the flow channel 12.

[0045] Optionally, in other embodiments, according to the actual design needs, the flow channel 12 can also be in communication with the bottom of the culture cavity 11.

[0046] In a possible embodiment, the culture cavity 11 is a through-hole structure with two orifices, and the bio-culture imaging integrated chip further comprises a transparent cover plate 3, which is used to cover the orifices of the through-hole structure to form a sealed bio-culture environment. The transparent cover plate 3 can be detachably arranged on the orifices, or fixedly arranged on the orifices. People can directly cover the transparent cover plate 3 on the orifices when the bio-culture imaging integrated chip is used, and take the transparent cover plate 3 away from the orifices after the bio-culture imaging integrated chip is used.

[0047] Optionally, during the process of collecting the image of the biological sample in the culture cavity 11 by the collecting device, the collecting device can be attached to one of the holes of the through-hole structure, and the transparent cover plate 3 can be attached to the other hole of the through-hole structure. The through-hole structure can be a through-hole structure that penetrates from top to bottom, and the two holes of the through-hole structure can include an upper hole and a lower hole. The collecting device can be attached to the lower hole of the through-hole structure, and the transparent cover plate 3 can be attached to the upper hole of the through-hole structure.

[0048] In a possible embodiment, the microfluidic structure 1 can be made of a material including polydimethylsiloxane (PDMS). The PDMS material can be a material prepared by using a prepolymer and a cross-linking agent as raw materials, by light curing, and in a mold with low surface roughness.

[0049] Optionally, the main component of the prepolymer can be a PDMS prepolymer (such as a siloxane oligomer), which can provide a flexible chain structure as an uncrosslinked liquid base material. The cross-linking agent can be a cross-linking agent containing hydrogen siloxane or a platinum catalyst, which can react with active groups such as vinyl groups in the prepolymer to initiate cross-linking and curing (vulcanization reaction).

[0050] In the production process of PDMS, the mixing ratio between the prepolymer and the cross-linking agent can be adjusted, and the prepolymer and the cross-linking agent can be mixed in a mixing ratio of 5:1-10:1. In actual operation, people can also adjust the mixing ratio between the prepolymer and the cross-linking agent according to actual production and life needs.

[0051] In a possible embodiment, the collecting device includes a complementary metal oxide semiconductor (CMOS) image sensor, or the collecting device includes a charge-coupled device (CCD) sensor. Both the CMOS image sensor and the CCD sensor can be used to collect images of biological samples, and people can replace the CMOS sensor and the CCD sensor according to actual production and life needs.

[0052] In a specific embodiment, during the process of using the CMOS sensor to collect images, the CMOS sensor is more cost-effective, and the chip made of the CMOS sensor has a lower cost. People can reduce production costs by using the CMOS image sensor as the collecting device.

[0053] Please refer to Figure 3 and Figure 4In another aspect, the application discloses a three-dimensional imaging system, which comprises any one of the above biological culture imaging integrated chips, at least one illumination lamp strip 4 and an electric control system. The illumination lamp strip 4 can be used to illuminate the culture cavity 11 on the biological culture imaging integrated chip, and the electric control system can be electrically connected with the biological culture imaging integrated chip and the illumination lamp strip 4.

[0054] Optionally, each illumination lamp strip 4 comprises at least two illumination devices 41, and each two adjacent illumination devices 41 in the at least two illumination devices 41 are spaced apart by a preset angle. The illumination device 41 can be any illumination device capable of illuminating the culture cavity, such as an LED lamp or an incandescent lamp, and the type of the illumination device 41 can be adjusted according to actual production and life needs.

[0055] Optionally, the illumination device 41 can be fixedly arranged on the illumination lamp strip 4, and each two adjacent illumination devices 41 in the at least two illumination devices 41 can be spaced apart by a fixed preset angle. Alternatively, the illumination device 41 can be detachably and adjustably arranged on the illumination lamp strip 4, and each two adjacent illumination devices 41 in the at least two illumination devices 41 can be spaced apart by an adjustable preset angle. The number and position of the illumination device 41 on the illumination lamp strip 4 can be adjusted according to actual observation needs. Each illumination device 41 can be arranged towards the culture cavity 11, and each illumination device 41 can illuminate the culture cavity 11.

[0056] Optionally, the electric control system can comprise a control circuit module composed of a Raspberry Pi, a single-chip microcomputer, an FPGA (Field Programmable Gate Array), a jetson nano or any other electronic device having a control function. The electric control system can be used to turn on or turn off the illumination device 41 on the illumination lamp strip 4, and can also be used to perform three-dimensional modeling of the biological sample in the culture cavity 11 based on the collected data of the biological culture imaging integrated chip.

[0057] During use of the three-dimensional imaging system, each illumination device 41 on each illumination lamp strip 4 is turned on and turned off in sequence, so that the image acquisition unit 2 can perform image acquisition when each illumination device 41 illuminates the culture cavity 11, until each illumination device 41 on each illumination lamp strip 4 is turned on and turned off for one round. The electric control system can establish a three-dimensional model of the biological sample based on the acquisition results of the image acquisition unit 2. Figure 4 A historical modeling schematic diagram for establishing a three-dimensional model for the electric control system.

[0058] In this embodiment, at least one lighting lamp strip 4 can perform multi-angle and multi-view imaging on the biological sample, thereby meeting the multi-angle lighting needs of the biological sample. At the same time, this embodiment can realize parallel processing of image acquisition, data transmission and three-dimensional model establishment, that is, in the process of transmitting current period data and establishing a model by the electric control system, the image acquisition unit 2 can acquire image data of the biological sample in the next period, thereby ensuring that the system can realize long-term, uninterrupted continuous imaging. The three-dimensional model of the biological sample can be established based on the lensless holographic three-dimensional imaging construction method provided in the patent CN114967397B.

[0059] In a possible embodiment, the three-dimensional imaging system includes two arc-shaped lighting lamp strips 4, and the image acquisition unit 2 is located at the bottom of the biological culture and imaging integrated chip. The two lighting lamp strips 4 can be orthogonally arranged above the biological culture and imaging integrated chip, and the two lighting lamp strips 4 can form a 90° angle.

[0060] In other embodiments, the arc-shaped lighting lamp strip 4 can also be replaced by a ring-shaped lighting lamp strip 4, and the biological culture and imaging integrated chip can be located at the center of the ring-shaped lighting lamp strip 4. However, the array type integrated arrangement of the ring-shaped lighting lamp strip 4 is more difficult, and the overall volume of the system is more difficult to compress. The embodiments of the present application prefer to use the arc-shaped lighting lamp strip 4.

[0061] Optionally, 41 lighting devices 41 can be uniformly arranged on each arc-shaped lighting lamp strip 4, and the interval angle between every two adjacent lighting devices 41 can be 6°. The lighting device 41 can be an LED lamp bead.

[0062] In a possible embodiment, the arc-shaped lighting lamp strip 4 can be a circular arc lighting lamp strip 4, and the biological culture and imaging integrated chip is located at the arc center position of the two arc-shaped lighting lamp strips 4, that is, the two circular arc lighting lamp strips 4 can be concentric, and the biological culture and imaging integrated chip is located at the center of the two circular arc lighting lamp strips 4. Therefore, the interval distance between each lighting device 41 on the circular arc lighting lamp strip 4 and the biological culture and imaging integrated chip is limited to a fixed value, thereby realizing more accurate image acquisition of the biological sample.

[0063] Optionally, the working distance between each lighting device 41 on the circular arc lighting lamp strip 4 and the biological culture and imaging integrated chip can be set to 10 centimeters. This distance helps to make the incident light of the lighting device 41 approximate to parallel plane light, thereby being able to optimize the imaging effect.

[0064] In a possible embodiment, each lighting device 41 is provided with a shielding member, which can be a circular mask structure arranged on the outer surface of the lighting device 41, the circular mask structure being capable of precisely shielding the angle range of the light beam emitted by the lighting device 41, the shielding member being used to limit the light angle of the lighting device 41 so as to concentrate the illumination of the culture cavity 11, thereby enabling the spot illumination of the biological sample in the culture cavity 11.

[0065] It should be noted that the above-mentioned sequence of the embodiments of the present application is merely for description, and does not represent the advantages and disadvantages of the embodiments. Moreover, the above-mentioned description is made for specific embodiments of the present application. Other embodiments are within the scope of the appended claims. In some cases, the actions or steps recited in the claims can be performed in a different order than the order in which they are recited and still achieve desirable results. In addition, the processes depicted in the figures do not necessarily require the particular order shown, or sequential order, to achieve the desired results. In certain implementations, multitasking and parallel processing can be advantageous.

[0066] Each of the embodiments in the present specification is described in a progressive manner, and the same or similar parts between the embodiments can be mutually referred to. Each of the embodiments focuses on the difference from other embodiments. In particular, for the device embodiments, since they are basically similar to the method embodiments, the description is relatively simple, and the relevant parts can be referred to the part of the description of the method embodiments.

Claims

1. A bio-culture imaging integrated chip, characterized in that, The microfluidic structure and the image acquisition part are included; The microfluidic structure is provided with a culture cavity; the culture cavity is a hole structure; the culture cavity is used for accommodating a biological sample; The image acquisition part includes an acquisition device; the acquisition device is arranged at the aperture of the culture cavity; the acquisition device is used for acquiring the image of the biological sample in the culture cavity.

2. The bio-culturing and imaging integrated chip according to claim 1, wherein, The microfluidic structure is provided with at least one flow channel; one end of the flow channel is communicated with the culture cavity, and the other end of the flow channel is communicated with the outside world; The biological sample can flow into or out of the culture cavity through the flow channel.

3. The bio-culturing and imaging integrated chip according to claim 2, wherein, The flow channel is communicated with the top of the culture cavity.

4. The bio-culturing and imaging integrated chip according to claim 2, wherein, The culture cavity is a through-hole structure with two apertures; The biological culture imaging integrated chip further includes a transparent cover plate; During the process that the acquisition device acquires the image of the biological sample in the culture cavity, the acquisition device is arranged at one aperture of the through-hole structure, and the transparent cover plate is arranged at the other aperture of the through-hole structure.

5. The bio-culturing and imaging integrated chip according to claim 1, wherein, The microfluidic structure is made of a material including polydimethylsiloxane; The polydimethylsiloxane material is a material prepared by using a prepolymer and a crosslinking agent as raw materials, through light curing, and in a mold with low surface roughness.

6. The bio-culturing and imaging integrated chip according to claim 5, wherein, The acquisition device includes a complementary metal oxide semiconductor image sensor, or the acquisition device includes a charge coupled device sensor.

7. A three-dimensional imaging system characterized by, The biological culture imaging integrated chip, at least one illumination lamp strip, and an electric control system are included; Each illumination lamp strip includes at least two illumination devices; each two adjacent illumination devices in the at least two illumination devices are spaced apart by a preset angle; each illumination device is arranged towards the culture cavity; During the use of the three-dimensional imaging system, each illumination device on each illumination lamp strip is sequentially turned on and turned off, so that the image acquisition part can acquire images when each illumination device illuminates the culture cavity; The electric control system can establish a three-dimensional model of the biological sample through the acquisition results of the image acquisition part.

8. The three-dimensional imaging system of claim 7, wherein, Two arc-shaped illumination lamp strips are included; The image acquisition part is located at the bottom of the biological culture imaging integrated chip; the two illumination lamp strips are orthogonally arranged above the biological culture imaging integrated chip.

9. The three-dimensional imaging system of claim 8, wherein, The biological culture imaging integrated chip is located at the arc center position of the two arc-shaped illumination lamp strips.

10. The three-dimensional imaging system of claim 7, wherein, Each illumination device is provided with a shielding piece; The shielding piece is used to limit the light angle of the illumination device so as to concentrate the illumination of the illumination device on the culture cavity.

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