Optical imaging sensor and biological reaction system

Abstract

The present disclosure relates to an optical imaging sensor and a biological reaction system. The optical imaging sensor comprises: an illumination assembly configured to generate an illumination beam in a first direction; a detection assembly configured to receive and detect an imaging beam formed from the illumination beam and in a second direction, wherein the first direction and the second direction intersect each other; and a housing configured to accommodate the illumination assembly and the detection assembly, wherein the housing comprises a window portion, and the window portion is located on an emergent light path of the illumination assembly and on an incident light path of the detection assembly, so that the illumination beam and the imaging beam can be transmitted through the window portion.

Description

Optical imaging sensors and bio-response systems

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application is based on and claims priority to the Chinese patent application with Chinese application number 202310903956.1 and application date July 21, 2023. The disclosed content of the Chinese patent application is hereby introduced as a whole into this application. Technical Field

[0003] The present disclosure relates to the field of optical detection technology, and in particular to an optical imaging sensor and a biological reaction system. Background Art

[0004] In research and production in biological-related fields, it is often necessary to monitor biological reaction processes, such as cell culture. However, current monitoring methods are usually ex situ and offline, that is, samples are extracted from the reaction vessel and then analyzed. This monitoring method is difficult to reflect the true state of the biological reaction in real time, nor can it provide continuous control signals to help achieve automated control. In addition, contamination may be introduced during the sampling process. Therefore, there is a need for improved monitoring of biological reactions.

[0005] Summary of the Invention

[0006] One of the objectives of the present disclosure is to provide an optical imaging sensor and a biological response system.

[0007] According to a first aspect of the present disclosure, there is provided an optical imaging sensor, comprising:

[0008] an illumination assembly configured to generate an illumination beam along a first direction;

[0009] a detection component configured to receive and detect an imaging light beam formed from the illumination light beam and along a second direction, wherein the first direction and the second direction intersect each other; and

[0010] The shell is configured to accommodate the lighting component and the detection component, and the shell includes a window portion, which is located on the outgoing light path of the lighting component and on the incident light path of the detection component, so that the illumination light beam and the imaging light beam can be transmitted through the window portion.

[0011] In some embodiments, the lighting assembly includes:

[0012] An illumination light source is configured to generate an initial light beam.

[0013] In some embodiments, the lighting assembly further comprises:

[0014] A beam shaping component is provided on the outgoing light path of the illumination light source, and the beam shaping component is configured to shape the light beam to generate an illumination beam with a preset shape.

[0015] In some embodiments, the beam shaping assembly comprises:

[0016] an aperture, the aperture being provided on an outgoing light path of the illumination light source, and comprising a light-shielding screen and a light-transmitting portion provided in the light-shielding screen, wherein the light-transmitting portion is rectangular; and

[0017] a cylindrical lens group, the cylindrical lens group being arranged on an exit light path of the aperture;

[0018] The cylindrical lens group and the aperture are configured to shape the light beam to generate a substantially sheet-shaped illumination light beam, and the plane where the illumination light beam is located is substantially perpendicular to the second direction.

[0019] In some embodiments, the cylindrical lens group includes a first cylindrical lens, wherein a convex surface of the first cylindrical lens is arranged to face the aperture.

[0020] In some embodiments, when the illumination light source is an incoherent light source, the minimum thickness Z of the illumination light beam in the second direction satisfies Z=1.4*f*λ / (2*D), and the effective length L of the illumination light beam in the first direction satisfies L=2*n*π*Z 2 / λ, and the effective width of the illumination light beam in the third direction satisfies W=D'*NA, wherein the third direction is perpendicular to both the first direction and the second direction, f is the focal length of the cylindrical lens group, λ is the wavelength of the illumination light beam, D is the side length of the light-transmitting portion in the second direction, D' is the side length of the light-transmitting portion in the third direction, n is the refractive index of the medium in which the illumination light beam is transmitted, and NA is the numerical aperture of the illumination assembly.

[0021] In some embodiments, when the illumination light source is a coherent light source, the minimum thickness Z of the illumination light beam in the second direction satisfies Z=2*λ / (π*NA), and the effective length L of the illumination light beam in the first direction satisfies L=2*n*λ / (π*NA 2 ), and the effective width of the illumination light beam in the third direction satisfies W=D'*NA, wherein the third direction is perpendicular to both the first direction and the second direction, λ is the wavelength of the illumination light beam, NA is the numerical aperture of the illumination assembly, n is the refractive index of the medium in which the illumination light beam is transmitted, and D' is the side length of the light-transmitting portion in the third direction.

[0022] In some embodiments, the iris includes an adjustable iris, and the light transmission range of the light-transmitting portion of the adjustable iris can be changed.

[0023] In some embodiments, the lighting assembly further comprises:

[0024] A collimating lens group is provided on the outgoing light path of the beam shaping component and is configured to collimate the light beam to generate a collimated illumination beam.

[0025] In some embodiments, the detection component includes:

[0026] an imaging lens group configured to receive the imaging light beam; and

[0027] An optical detector is provided on an exit light path of the imaging lens group and is configured to sense the imaging light beam.

[0028] In some embodiments, the imaging lens assembly includes:

[0029] objective lens; and

[0030] A tube lens is provided between the objective lens and the optical detector.

[0031] In some embodiments, the detection component further comprises:

[0032] A filter is provided on the incident light path of the optical detector, and the filter is configured to filter out a portion of the imaging light beam in a preset wavelength band and allow a portion of the imaging light beam in other wavelength bands to pass.

[0033] In some embodiments, the illumination assembly is positioned such that the illumination beam covers at least a focal plane of the detection assembly.

[0034] In some embodiments, the window portion is formed of a light-transmitting hard material or a light-transmitting flexible material.

[0035] In some embodiments, other portions of the housing except the window portion are formed of a non-light-transmitting material.

[0036] According to a second aspect of the present disclosure, there is provided a bioreactor system comprising:

[0037] a reaction vessel, wherein at least a portion of the reaction vessel is formed of a light-transmitting material; and

[0038] The optical imaging sensor as described above, wherein the window portion of the optical imaging sensor is configured to be in contact with a portion of the reaction container formed of a light-transmitting material, so as to monitor a biological reaction in the reaction container.

[0039] In some embodiments, the bioreactor system further comprises:

[0040] a memory having instructions stored therein; and

[0041] A processor is communicatively connected to the optical imaging sensor and the memory, and when the instructions are executed by the processor, performs image processing on the imaging image generated by the optical imaging sensor.

[0042] Other features and advantages of the present disclosure will become apparent from the following detailed description of exemplary embodiments of the present disclosure with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the present disclosure and, together with the description, serve to explain the principles of the present disclosure.

[0044] The present disclosure can be more clearly understood from the following detailed description with reference to the accompanying drawings, in which:

[0045] FIG1 shows a schematic structural diagram of a bioreactor system according to a specific embodiment of the present disclosure;

[0046] FIG2 is a schematic diagram showing the bioreactor system in FIG1 in a working state;

[0047] FIG3 shows a partially enlarged schematic diagram of the bioreactor system in FIG1 in a working state;

[0048] FIG4 shows a schematic structural diagram of a bioreactor system according to another specific embodiment of the present disclosure;

[0049] FIG5 shows a schematic structural diagram of an optical imaging sensor according to an exemplary embodiment of the present disclosure;

[0050] FIG6 shows a schematic structural diagram of an aperture according to a specific embodiment of the present disclosure;

[0051] FIG7 is a schematic diagram showing an illumination beam generated when a cylindrical lens group including a first cylindrical lens is used according to a specific embodiment of the present disclosure;

[0052] FIG8 shows a schematic cross-sectional view of the illumination beam in FIG7 on the xz plane;

[0053] FIG9 shows a schematic cross-sectional view of the illumination beam in FIG7 on the xy plane;

[0054] FIG10 shows an imaging view of cells in a bioreactor system according to a specific application example of the present disclosure;

[0055] FIG11 shows a graph showing changes in total cell density over time in a bioreactor system according to a specific application example of the present disclosure;

[0056] FIG12 shows a graph showing changes in live cell density over time in a bioreactor system according to a specific application example of the present disclosure;

[0057] FIG13 shows a graph showing changes in cell viability over time in a bioreactor system according to a specific application example of the present disclosure;

[0058] FIG14 shows a curve diagram showing changes in cell diameter over time in a bioreactor system according to a specific application example of the present disclosure.

[0059] Note that in the embodiments described below, the same reference numerals are sometimes used in common across different drawings to denote the same parts or parts having the same functions, and their repeated descriptions are omitted. In this specification, similar reference numerals and letters are used to denote similar items. Therefore, once an item is defined in one drawing, it need not be further discussed in subsequent drawings.

[0060] To facilitate understanding, the positions, sizes, and ranges of various structures shown in the drawings and the like may not always represent actual positions, sizes, and ranges. Therefore, the disclosed invention is not limited to the positions, sizes, and ranges disclosed in the drawings and the like. Furthermore, the drawings are not necessarily drawn to scale, and some features may be exaggerated to illustrate details of specific components. DETAILED DESCRIPTION

[0061] Various exemplary embodiments of the present disclosure will now be described in detail with reference to the accompanying drawings. It should be noted that unless otherwise specifically stated, the relative arrangement of components and steps, numerical expressions and numerical values ​​set forth in these embodiments do not limit the scope of the present disclosure.

[0062] The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the present disclosure, its application, or use. That is, the devices and methods described herein are presented in an exemplary manner to illustrate various embodiments of the present disclosure and are not intended to be limiting. Those skilled in the art will appreciate that these descriptions are merely illustrative of exemplary ways that may be used to implement the present disclosure, and are not intended to be exhaustive.

[0063] Technologies, methods and equipment known to ordinary technicians in the relevant art may not be discussed in detail, but where appropriate, such technologies, methods and equipment should be considered part of the authorization specification.

[0064] In research and production in the biotechnology field, monitoring of reaction processes, such as biological culture, is often required. Currently, culture vessels such as culture dishes, shake flasks, reactors, and disposable bio-culture bags (e.g., cell culture bags) are commonly used to culture cells or microorganisms. Disposable bio-culture bags are particularly popular in large-scale culture applications such as biopharmaceuticals and synthetic biology.

[0065] During the culture process, the accuracy of detecting parameters such as the number, viability and density of cells will directly affect the optimization of culture conditions, the control of production conditions and the quality of the final biopharmaceutical product. At present, the main detection method is ectopic and offline, that is, samples are extracted from a reaction vessel (e.g., a reaction tank or a culture bag), and then the extracted samples are subjected to cell analysis. For example, cell sensors based on dielectric spectrum or capacitance can be used to indirectly measure some parameters. In this measurement method, the electrical signals generated by the cells polarized by the radio frequency electric field can be collected in advance, and a correlation relationship between the electrical signals and parameters such as the density of living cells can be established. During the measurement process, offline detection equipment can be used to obtain electrical signal measurement values, and the relevant parameters of the sample can be obtained based on the pre-established correlation relationship. However, the measurement method based on the above-mentioned cell sensor can only roughly obtain information related to the number and density of living cells, with low measurement accuracy, and cannot measure parameters such as the total number of cells (including living cells and dead cells) or total cell density. In addition, the above-mentioned indirect measurement scheme is also difficult to monitor the cell growth status continuously and in real time. At the same time, the process of extracting samples may also introduce the risk of cross-contamination, and the process and time spent on sample preparation may also affect the accuracy and real-time nature of the monitoring results.

[0066] In order to solve the above problems, the present disclosure proposes an optical imaging sensor and a bioreaction system, wherein the optical imaging sensor can perform real-time optical imaging of microorganisms or plant and animal cells in the original reaction environment, thereby realizing continuous and real-time monitoring. Furthermore, it can be combined with, for example, image processing software to realize automated processing of imaging images, extract or calculate the required parameters therefrom, thereby realizing real-time analysis of microorganisms or plant and animal cells in the original reaction environment. The optical imaging sensor disclosed in the present disclosure can be used in a variety of reaction containers such as cell culture dishes, T bottles, cell culture bags, and can be used to obtain continuous and real-time status data of cells in the original culture environment, such as cell density, cell viability, cell diameter / volume, cell classification and grouping, cell performance (for example, cytokine secretion), cell characteristics (for example, surface markers or genetic material), cell growth rate, and cell distribution at different levels in the reaction container.

[0067] In exemplary embodiments of the present disclosure, an optical imaging sensor for imaging a sample to be tested can be provided separately or integrated with, for example, a reaction vessel for culturing cells, to form at least a portion of a bioreactor system. As shown in Figures 1 to 4, in some embodiments of the present disclosure, the bioreactor system can include a reaction vessel 200 and an optical imaging sensor 100. The reaction vessel 200 can also be provided with a stirring portion 210 or the like to agitate the liquid containing the biological particles 900 during the reaction process to achieve a better reaction effect. At least a portion of the reaction vessel 200 can be formed from a light-transmitting material so that the window portion 131 of the optical imaging sensor 100 can abut against the light-transmitting portion of the reaction vessel 200, thereby allowing the illumination beam generated by the optical imaging sensor 100 to illuminate the sample in the reaction vessel 200. The imaging beam generated by the interaction of the illumination beam with the biological particles 900 in the sample can then pass through the reaction vessel 200 and return to the optical imaging sensor 100 for imaging. In one specific example, the entire reaction vessel 200 can be formed of a light-transmitting material, so that the window portion 131 of the optical imaging sensor 100 can be placed at most locations on the reaction vessel 200 to monitor biological reactions at desired locations. In other specific examples, only a portion of the reaction vessel 200 is formed of a light-transmitting material, while the other portions can be formed of an opaque material. This minimizes the entry of unnecessary ambient light into the reaction vessel 200 while ensuring the monitoring effect of the optical imaging sensor 100, thereby reducing interference with the biological reactions therein. For example, the light-transmitting portion of the reaction vessel 200 can be a long strip extending along its longitudinal axis. In this way, the window portion 131 of the optical imaging sensor 100 can move along the longitudinal axis of the reaction vessel 200 to monitor biological reactions at multiple levels in the reaction vessel 200.

[0068] In the specific examples shown in Figures 1 to 3 , when the bioreactor system is in operation, the reaction vessel 200 can be placed horizontally, the optical imaging sensor 100 can be disposed at the bottom of the reaction vessel 200, and the window 131 of the optical imaging sensor 100 can be in contact with the light-transmitting portion of the reaction vessel 200. In the specific example shown in Figure 4 , when the bioreactor system is in operation, the reaction vessel 200 can be placed vertically, the optical imaging sensor 100 can be disposed on the side of the reaction vessel 200, for example, attached to the sidewall of the reaction vessel 200, and the window 131 of the optical imaging sensor 100 can be in contact with the light-transmitting portion of the reaction vessel 200.

[0069] In some embodiments, as shown in FIG1 , the bioreactor system may further include a memory 300 and a processor 400. For example, the memory 300 and processor 400 may be part of a computer device that is communicatively connected to the optical imaging sensor 100. In some embodiments, the computer device may be independently provided; in other embodiments, the computer device may also be part of the bioreactor system. Instructions may be stored in the memory 300. The processor 400 may be communicatively connected to the optical imaging sensor 100 and the memory 300. When the instructions are executed by the processor 400, image processing may be performed on the images generated by the optical imaging sensor 100, such as cell counting and viability analysis based on the images. In some embodiments, the memory 300 may also be used to store raw image data from the optical imaging sensor 100, various intermediate data involved in the image processing process, and image processing result data. Furthermore, in some embodiments, the memory 300 may also be directly communicatively connected to the optical imaging sensor 100, without limitation.

[0070] The memory 300 may be a volatile memory or a non-volatile memory, or may include both volatile and non-volatile memories. The non-volatile memory may be a read-only memory (ROM), a programmable read-only memory (PROM), an erasable programmable read-only memory (EPROM), an electrically erasable programmable read-only memory (EEPROM), or a flash memory. The volatile memory may be a random access memory (RAM), which is used as an external cache. By way of example but not limitation, many forms of RAM are available, such as static random access memory (SRAM), dynamic random access memory (DRAM), synchronous dynamic random access memory (SDRAM), double data rate synchronous dynamic random access memory (DDRSDRAM), enhanced synchronous dynamic random access memory (ESDRAM), synchronous linked dynamic random access memory (SLDRAM), and direct memory bus random access memory (DR RAM). It should be noted that the memory 300 described herein is intended to include, but is not limited to, these and any other suitable types of memory.

[0071] The processor 400 can be an integrated circuit chip with signal processing capabilities. The above-mentioned processor can be a general-purpose processor, a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA) or other programmable logic device, a discrete gate or transistor logic device, or a discrete hardware component. The various methods, steps, and logic block diagrams mentioned in the embodiments of the present disclosure can be implemented or executed. The general-purpose processor can be a microprocessor or any conventional processor, etc., and can be an X86 architecture or an ARM architecture, etc.

[0072] In an exemplary embodiment of the present disclosure, as shown in Figures 3 and 5, the optical imaging sensor 100 may include an illumination assembly 110, a detection assembly 120, and a housing 130. The illumination assembly 110 may be configured to generate an illumination beam along a first direction (i.e., the direction of the x-axis in Figure 5); the detection assembly 120 may be configured to receive and detect an imaging beam formed from the illumination beam along a second direction (i.e., the direction of the z-axis in Figure 5), where the first direction and the second direction may intersect or be perpendicular to each other; and the housing 130 may be configured to accommodate the illumination assembly 110 and the detection assembly 120, and the housing 130 may include a window portion 131, which may be located on an outgoing light path of the illumination assembly 110 and an incident light path of the detection assembly 120, so that the illumination beam and the imaging beam can be transmitted through the window portion 131.

[0073] In some embodiments, as shown in FIG5 , the illumination assembly 110 may include an illumination light source 111, which may be configured to generate an initial light beam. The initial light beam may be an incoherent light beam or a coherent light beam, and accordingly, the illumination light beam irradiated onto the sample to be tested may be an incoherent light beam or a coherent light beam. In a specific example, elastic scattering may occur between the illumination light beam and the biological particles 900 in the sample to be tested, and the morphology and other aspects of the sample to be tested may be observed based on the resulting scattered light. In this case, the illumination light beam may be an incoherent light beam, and accordingly, the illumination light source 111 may be an incoherent light source such as a light-emitting diode (LED), which may generate, for example, white light for easier observation. Furthermore, particularly when the illumination light source 111 is an incoherent light source, it may generally have a smaller size, and accordingly, the entire optical imaging sensor 100 may also have a smaller size. For example, when an incoherent light source is provided, the characteristic dimensions (e.g., length, width, or thickness) of the housing of the optical imaging sensor 100 may be less than 10 cm. In another specific example, the illumination light beam can be used to excite the fluorescence signal of the biological particles 900 in the sample to be tested, thereby achieving fluorescence detection of the sample to be tested based on the generated fluorescence signal to obtain relevant information about the biological particles 900. In this case, to improve the excitation effect, the illumination light beam can be a coherent light beam in the desired wavelength band. Accordingly, the illumination light source 111 can include a coherent light source such as a laser, or can include an optical fiber for transmitting the coherent light beam. It will be understood that in some embodiments, the illumination light source 111 can also be a hybrid light source that can generate both incoherent and coherent light beams, or can include multiple illumination light sources 111 to switch to the corresponding illumination light beam as needed.

[0074] In some embodiments, the lighting assembly 110 may further include a beam shaping assembly. This beam shaping assembly may be disposed on the outgoing light path of the illumination light source 111 and may be configured to shape the light beam to produce an illumination beam having a predetermined shape, thereby achieving a better monitoring effect. In a specific example, as shown in FIG5 , the beam shaping assembly may include an aperture 112 and a cylindrical lens group 113. The aperture 112 and the cylindrical lens group 113 may be configured to shape the light beam to produce a substantially sheet-shaped illumination beam, and the plane of the illumination beam may be perpendicular or substantially perpendicular to the second direction.

[0075] In the specific example shown in FIG5 , the aperture 112 can be disposed in the outgoing light path of the illumination light source 111. As shown in FIG6 , the aperture 112 can include a light shield 112a and a light-transmitting portion 112b defined within the light shield 112a. The shape of the light-transmitting portion 112b can affect the cross-sectional shape of the illumination beam in the yz plane (where the y direction is perpendicular to both the x and z directions) shown in FIG5 . In some embodiments, the light-transmitting portion 112b can be rectangular. In a specific example, the light-transmitting portion 112b can be square. To ensure substantially uniform illumination within the observation field of the detection assembly 120, thereby enabling better observation of the sample under test and more convenient calculation of required parameters, the effective width of the illumination beam (the width of the portion of the beam where the light intensity exceeds a certain threshold to provide sufficient illumination resolution) can be greater than the field of view of the detection assembly 120 (or the objective lens 123 within the detection assembly 120, as described below), allowing the illumination beam to be distributed across the entire field of view. Furthermore, in some embodiments, the aperture 112 may include an adjustable aperture, the light transmission range of the light-transmitting portion of the adjustable aperture can be changed to change the shape of the illumination beam as needed, such as changing the effective width of the illumination beam to achieve a desired illumination range.

[0076] In the specific example shown in FIG5 , cylindrical lens group 113 can be disposed on the exit optical path of aperture 112. As described above, cylindrical lens group 113 and aperture 112 can, together, shape the initial light beam into a sheet-shaped illumination beam, with the normal direction of the sheet beam substantially along the second direction, and the plane of the sheet beam extending in the xy plane. It should be noted that the sheet-shaped beam described herein refers to a beam whose thickness in the z direction is much smaller (e.g., more than 10 times smaller) than its effective length in the x direction (the length of the portion of the beam where the light intensity exceeds a certain threshold to provide sufficient illumination resolution), and whose thickness is much smaller than its effective width in the y direction.

[0077] The cylindrical lens assembly 113 can be configured in a variety of ways to produce a desired sheet-shaped light beam. Furthermore, the one or more cylindrical lenses employed in the cylindrical lens assembly 113 can each be a single-piece cylindrical lens, or a multi-piece apochromatic cylindrical lens, to make the effective illumination portion of the sheet-shaped light beam more uniform and avoid the effects of chromatic aberration on optical observation.

[0078] In a specific example, as shown in FIG5 , the cylindrical lens group 113 may include a first cylindrical lens, wherein the convex surface of the first cylindrical lens may be disposed facing the aperture 112. In this case, the first cylindrical lens may generate a sheet-shaped light beam as shown in FIG7 to FIG9 , wherein the edge of the sheet-shaped light beam extending along the x-direction may have a certain curvature.

[0079] It is understood that the geometric dimensions of the generated illumination beam are related to the corresponding components in the illumination assembly 110 and the parameters of the medium in which the illumination beam is transmitted. In some embodiments, the thickness and length of the illumination beam can be calculated based on the diffraction limit. For example, when the illumination light source 111 is an incoherent light source, the minimum thickness Z of the illumination beam 119 in the second direction satisfies Z = 1.4 * f * λ / (2 * D), and the effective length L of the illumination beam 119 in the first direction satisfies L = 2 * n * π * Z 2 / λ, and the effective width of the illumination beam 119 in the third direction (i.e., the direction of the y-axis in FIG5 ) satisfies W=D'*NA, wherein the third direction may be a direction perpendicular to both the first direction and the second direction, f is the focal length of the cylindrical lens group 113, λ is the wavelength of the illumination beam 119, D is the side length of the light-transmitting portion 112b in the second direction (i.e., the direction of the z-axis in FIG5 ), D' is the side length of the light-transmitting portion 112b in the third direction (i.e., the direction of the y-axis in FIG5 ), n is the refractive index of the medium in which the illumination beam 119 is transmitted (in the present disclosure, the illumination beam may be mainly transmitted in a vacuum or air, so the refractive index n may be 1), and NA is the numerical aperture of the illumination assembly 110 (or the collimating lens group 114 described below). In a specific example, the thickness of the illumination beam 119 may be The positions are determined to correspond to the two end points of the effective length of the illumination beam.

[0080] In other embodiments, when the illumination light source 111 is a coherent light source, the minimum thickness Z of the illumination light beam 119 in the second direction satisfies Z=2*λ / (π*NA), and the effective length L of the illumination light beam 119 in the first direction satisfies L=2*n*λ / (π*NA 2), and the effective width of the illumination beam 119 in the third direction satisfies W=D'*NA, where λ is the wavelength of the illumination beam 119, NA is the numerical aperture of the illumination assembly 110 (or the collimating lens group 114 described below), n is the refractive index of the medium in which the illumination beam 119 is transmitted (in the present disclosure, the illumination beam can be mainly transmitted in a vacuum or air, so the refractive index n can be 1), and D' is the side length of the light-transmitting portion 112b in the third direction (i.e., the direction of the y-axis in FIG5 ). Similarly, the thickness of the illumination beam 119 can be The positions of are determined to correspond to the two end points of the effective length of the illumination beam 119.

[0081] In some embodiments, in order to further reduce the thickness of the illumination beam 119 so that the optical imaging sensor 100 can have a finer axial resolution in the thickness direction during layer-by-layer analysis of the sample to be tested (the axial resolution is related to the thickness of the illumination beam and the numerical aperture of the objective lens 123 in the detection assembly 120), or to make the energy of the coherent light beam more concentrated to better excite the fluorescent signal in the sample to be tested, as shown in FIG5 , the illumination assembly 110 may further include a collimating lens group 114. The collimating lens group 114 may be disposed on the output light path of the beam shaping assembly, and the collimating lens group 114 may be configured to collimate the light beam to produce a collimated illumination beam, for example, to make the thickness of the illumination beam in the second direction thinner than the thickness in the absence of the collimating lens group 114. In some embodiments, the collimating lens group 114 may include one or more lenses.

[0082] Furthermore, as shown in FIG5 , the detection assembly 120 may include an imaging lens group and an optical detector 121. The imaging lens group may be configured to receive an imaging light beam, which may be from a layer of the sample to be tested that is illuminated by the illumination light beam. In some embodiments, the imaging lens group may include an objective lens 123 and a tube lens 122, with the tube lens 122 being disposed between the objective lens 123 and the optical detector 121. It is understood that in other embodiments, the imaging lens group for collecting the imaging light beam may also be configured in other ways, which are not limiting herein.

[0083] The optical detector 121 can be disposed on the outgoing light path of the imaging lens group, and the optical detector 121 can be configured to sense the imaging light beam, thereby forming a two-dimensional image of the sample to be tested (including an elastic scattered light image and / or a fluorescence signal image). In a specific example, the optical detector 121 can include a charge-coupled device (CCD) sensor or a complementary metal oxide semiconductor (CMOS) sensor. In addition, by scanning the sample to be tested along the second direction or by moving the entire optical imaging sensor 100 in the second direction, layer-by-layer imaging of the sample to be tested can be achieved, thereby obtaining tomographic images of the sample to be tested at different depths, thereby achieving three-dimensional information reconstruction of the sample to be tested.

[0084] In some embodiments, particularly when fluorescence imaging is performed, as shown in FIG5 , the detection assembly 120 may further include a filter 124. The filter 124 may be disposed in the incident light path of the optical detector 121. The filter 124 may be configured to filter out a portion of the imaging light beam within a predetermined wavelength band and pass portions of the imaging light beam within other wavelength bands (e.g., filtering out a portion of the imaging light beam within the wavelength band of the illumination light beam while allowing the fluorescence signal to pass). In this way, interference with the detection caused by light outside the wavelength band of the fluorescence signal can be reduced, thereby facilitating better detection of the sample to be tested.

[0085] In some embodiments, to achieve better imaging, as shown in FIG5 , the position of illumination assembly 110 can be set so that the illumination beam at least covers the focal plane of detection assembly 120, allowing detection assembly 120 to sense only the images of biological particles located near the focal plane of the imaging lens assembly. In other words, the positional relationship between illumination assembly 110 and detection assembly 120 ensures that the area illuminated by the sheet-like illumination beam is substantially located on the focal plane of detection assembly 120, thereby achieving the best imaging effect.

[0086] In some embodiments, as shown in Figures 3 and 5, the lighting assembly 110 and the imaging assembly 120 are both contained in the housing 130 of the optical imaging sensor 100, thereby forming a whole that is easy to carry. When it is necessary to monitor a biological reaction such as cell culture in a reaction vessel 200 such as a reaction bag, the optical imaging sensor 100 can be placed against the light-transmitting portion of the reaction vessel 200, wherein the window portion 131 of the housing 130 is opposite to the light-transmitting portion of the reaction vessel 200 so that the illumination beam and the imaging beam can be transmitted along the desired optical path. In some embodiments, the window portion 131 can be formed of a light-transmitting hard material (e.g., quartz). In this way, especially when monitoring a biological reaction in a reaction vessel 200 formed of a flexible material, the hard window portion 131 is conducive to making the area of ​​the reaction vessel 200 that abuts the optical imaging sensor 100 more flat, thereby improving the observation effect. In other embodiments, the window portion 131 can also be formed of a light-transmitting flexible material (e.g., polyethylene (PE)). In this way, the window portion 131 can conformally fit against the surfaces of reaction vessels 200 of various shapes to improve monitoring performance. Furthermore, in some embodiments, the housing 130, except for the window portion 131, can be formed of a non-light-transmitting material to prevent ambient light from entering the optical imaging sensor 100 and causing undesirable interference. In a specific example, the housing 130 can be formed by connecting several independent components, or the housing 130 can be integrally formed, without limitation herein.

[0087] Some specific application examples in optical imaging sensors and bioreaction systems are listed below.

[0088] In a specific application example, an optical imaging sensor can be used to monitor a series of parameters of CHO cells in a bioreactor system. Under batch feeding (FB) conditions, the growth process of CHO cells in a reaction vessel may include a logarithmic growth phase, a volume growth phase and a plateau phase, wherein the volume growth phase is between the logarithmic growth phase and the plateau phase. During the volume growth phase, the cells do not stop growing, but grow in a different way, that is, the increase in cell number is changed to the increase in single cell biomass, at which time the cell volume and diameter will increase. For example, during the volume growth phase, the cell diameter may increase from 17 μm to 23 μm. At the same time, the titer distribution of monoclonal antibodies in the cell volume increase process may be significantly higher than the titer distribution in the process where the cell volume is not increased. The optical imaging sensor proposed in the present disclosure can be used to monitor CHO cells in real time throughout the culture process. For example, Figure 10 shows an optical imaging photograph of cell 900 obtained during the culture process. Based on the optical imaging photographs shown in Figure 10, relevant image processing software can be used to extract or calculate information such as the change in the total cell density (TCD) of CHO cells over time (as shown in Figure 11, where the horizontal axis represents days and the vertical axis represents TCD), the change in the viable cell density (VCD) over time (as shown in Figure 12, where the horizontal axis represents days and the vertical axis represents VCD), the change in cell viability (Via) over time (as shown in Figure 13, where the horizontal axis represents days and the vertical axis represents Via), and the change in diameter (Dia) over time (as shown in Figure 14, where the horizontal axis represents days and the vertical axis represents Dia). Cell growth curves can also be drawn, providing timely and effective assistance to scientific researchers in optimizing and changing processes in antibody production.

[0089] Specifically, in the process of calculating the cell density, the field of view area of ​​the objective lens 123 in the detection assembly 120 can be used as the area. In the case where the depth of field of the objective lens 123 is greater than the thickness Z of the illumination beam, the effective volume is obtained by multiplying the thickness Z of the illumination beam by the field of view area, and the cell density is calculated based on the effective volume. Alternatively, in the case where the depth of field of the objective lens 123 is less than the thickness Z of the illumination beam, the effective volume can be obtained by multiplying the depth of field of the objective lens 123 by the field of view area, and the cell density in the sample to be tested is calculated based on the effective volume.

[0090] In the optical imaging sensor and biological reaction system disclosed in the present invention, an illumination light path and a detection light path that intersect or even are orthogonal to each other can be used, and a sheet-shaped illumination light beam can be used to illuminate the sample to be tested from the side, so that only the sample in the focal plane area (within the depth of field range) of the detection component is imaged. By detecting and imaging in the orthogonal direction, the influence of the out-of-focus background on the imaging quality can be effectively avoided. At the same time, it can have a high imaging contrast and naturally realize optical tomography to construct a three-dimensional model, greatly reducing the photobleaching and phototoxicity of the illumination light beam to the sample to be tested. Therefore, it is suitable for long-term imaging of living biological samples for several hours or even days, and there is no need for pre-modeling operations. For example, using the optical imaging sensor disclosed in the present invention, during the entire process of the biological reaction in the reaction vessel, the detection and statistics of parameters such as the counting, density, viability, volume, group classification and growth rate of biological samples can be achieved. In addition, the optical imaging sensor disclosed herein can be integrated into a housing, is small in size, low in cost, and easy to carry. When measurement is required, optical detection can be immediately achieved by simply placing the window of the optical imaging sensor against the light-transmitting portion of the reaction vessel. The measurement process is non-invasive and non-destructive, and does not require sampling or sample pretreatment, so it will not contaminate the reactants in the reaction vessel or interfere with the reaction process (e.g., cell culture, metabolism, etc.). Moreover, since there is no need to pre-stain the biological particles (e.g., using trypan blue staining to monitor cell activity, or using specific fluorescent reagents to monitor cell characteristics or surface markers, etc.), the time for pre-processing is saved, and the influence of the chemical toxicity of the reagent itself on the cell state and characteristic expression is avoided, thereby facilitating in-situ, real-time, direct, and continuous cell dynamic analysis.

[0091] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not limiting. Therefore, other examples of the exemplary embodiments may have different values.

[0092] The terms "front," "back," "top," "bottom," "above," "below," and the like, if any, in the specification and claims, are used for descriptive purposes and are not necessarily intended to describe invariant relative positions. It is understood that the terms so used are interchangeable under appropriate circumstances such that the embodiments of the disclosure described herein, for example, are capable of operation in other orientations than those illustrated or otherwise described herein.

[0093] As used herein, the word "exemplary" means "serving as an example, instance, or illustration," rather than as a "model" to be precisely copied. Any implementation described as exemplary is not necessarily to be construed as preferred or advantageous over other implementations. Furthermore, the present disclosure is not to be bound by any expressed or implied theory presented in the preceding technical field, background, summary, or detailed description.

[0094] As used herein, the word "substantially" is intended to include any minor variations due to design or manufacturing imperfections, device or component tolerances, environmental influences, and / or other factors. The word "substantially" also allows for deviations from a perfect or ideal condition due to parasitic effects, noise, and other practical considerations that may be present in actual implementations.

[0095] The above description may indicate elements or nodes or features that are "connected" or "coupled" together. As used herein, unless expressly stated otherwise, "connected" means that one element / node / feature is directly connected (or directly communicates) with another element / node / feature electrically, mechanically, logically, or otherwise. Similarly, unless expressly stated otherwise, "coupled" means that one element / node / feature can be mechanically, electrically, logically, or otherwise connected to another element / node / feature in a direct or indirect manner to allow interaction, even if the two features may not be directly connected. In other words, "coupled" is intended to encompass both direct and indirect connections of elements or other features, including connections utilizing one or more intermediate elements.

[0096] It should also be understood that when the term “include / comprises” is used in this document, it indicates the presence of the specified features, integers, steps, operations, units and / or components, but does not exclude the presence or addition of one or more other features, integers, steps, operations, units and / or components and / or their combinations.

[0097] Those skilled in the art will appreciate that the boundaries between the above-mentioned operations are merely illustrative. Multiple operations can be combined into a single operation, a single operation can be distributed among additional operations, and operations can be performed at least partially overlapping in time. Moreover, alternative embodiments can include multiple instances of specific operations, and the order of operations can be changed in various other embodiments. However, other modifications, variations, and replacements are also possible. Therefore, this specification and the accompanying drawings should be considered illustrative, not restrictive.

[0098] Although some specific embodiments of the present disclosure have been described in detail by way of examples, it should be understood by those skilled in the art that the above examples are for illustration only and are not intended to limit the scope of the present disclosure. The various embodiments disclosed herein may be combined in any manner without departing from the spirit and scope of the present disclosure. It should also be understood by those skilled in the art that various modifications may be made to the embodiments without departing from the scope and spirit of the present disclosure. The scope of the present disclosure is defined by the appended claims.

Claims

1. An optical imaging sensor, comprising: an illumination assembly configured to generate an illumination beam along a first direction; a detection component configured to receive and detect an imaging light beam formed from the illumination light beam and along a second direction, wherein the first direction and the second direction intersect each other; as well as The shell is configured to accommodate the lighting component and the detection component, and the shell includes a window portion, which is located on the outgoing light path of the lighting component and on the incident light path of the detection component, so that the illumination light beam and the imaging light beam can be transmitted through the window portion.

2. The optical imaging sensor according to claim 1, wherein: The lighting assembly comprises: An illumination light source is configured to generate an initial light beam.

3. The optical imaging sensor according to claim 2, wherein: The lighting assembly further comprises: A beam shaping component is arranged on the outgoing light path of the illumination light source, and the beam shaping component is configured to shape the light beam to generate an illumination beam with a preset shape.

4. The optical imaging sensor according to claim 3, wherein: The beam shaping assembly comprises: An aperture, the aperture being arranged on the outgoing light path of the illumination light source, and comprising a light-shielding screen and a light-transmitting portion provided in the light-shielding screen, wherein the light-transmitting portion is rectangular; and A cylindrical lens group, wherein the cylindrical lens group is arranged on the exit light path of the aperture; The cylindrical lens group and the aperture are configured to shape the light beam to generate a substantially sheet-shaped illumination light beam, and the plane where the illumination light beam is located is substantially perpendicular to the second direction.

5. The optical imaging sensor according to claim 4, wherein: The cylindrical lens group includes a first cylindrical lens, wherein a convex surface of the first cylindrical lens is arranged to face the aperture.

6. The optical imaging sensor according to claim 4, wherein: When the illumination light source is an incoherent light source, the minimum thickness Z of the illumination light beam in the second direction satisfies Z=1.4*f*λ / (2*D), and the effective length L of the illumination light beam in the first direction satisfies L=2*n*π*Z 2 / λ, and the effective width of the illumination light beam in the third direction satisfies W=D'*NA, wherein the third direction is perpendicular to both the first direction and the second direction, f is the focal length of the cylindrical lens group, λ is the wavelength of the illumination light beam, D is the side length of the light-transmitting portion in the second direction, D' is the side length of the light-transmitting portion in the third direction, n is the refractive index of the medium in which the illumination light beam is transmitted, and NA is the numerical aperture of the illumination component.

7. The optical imaging sensor according to claim 4, wherein: When the illumination light source is a coherent light source, the minimum thickness Z of the illumination light beam in the second direction satisfies Z=2*λ / (π*NA), and the effective length L of the illumination light beam in the first direction satisfies L=2*n*λ / (π*NA 2 ), and the effective width of the illumination light beam in the third direction satisfies W=D'*NA, wherein the third direction is perpendicular to both the first direction and the second direction, λ is the wavelength of the illumination light beam, NA is the numerical aperture of the illumination component, n is the refractive index of the medium in which the illumination light beam is transmitted, and D' is the side length of the light-transmitting portion in the third direction.

8. The optical imaging sensor according to claim 4, wherein: The diaphragm includes an adjustable diaphragm, and a light transmission range of a light transmission portion of the adjustable diaphragm can be changed.

9. The optical imaging sensor according to claim 4, wherein: The lighting assembly further comprises: A collimating lens group is arranged on the outgoing light path of the beam shaping component, and the collimating lens group is configured to collimate the light beam to generate a collimated illumination light beam.

10. The optical imaging sensor according to claim 1, wherein: The detection component comprises: an imaging lens group, the imaging lens group being configured to receive the imaging light beam; and An optical detector is disposed on an outgoing light path of the imaging lens group and is configured to sense the imaging light beam.

11. The optical imaging sensor according to claim 10, wherein: The imaging lens group comprises: objective lens; and A tube lens is arranged between the objective lens and the optical detector.

12. The optical imaging sensor according to claim 10, wherein: The detection component also includes: A filter is provided on the incident light path of the optical detector, and the filter is configured to filter out a portion of the imaging light beam in a preset wavelength band and pass a portion of the imaging light beam in other wavelength bands. Pass.

13. The optical imaging sensor according to claim 1, wherein: The illumination assembly is positioned such that the illumination beam covers at least a focal plane of the detection assembly.

14. The optical imaging sensor according to claim 1, wherein: The window portion is formed of a light-transmitting hard material or a light-transmitting flexible material.

15. The optical imaging sensor according to claim 1, wherein: The other parts of the housing except the window portion are formed of a non-light-transmitting material.

16. A bioreactor system comprising: A reaction container, wherein at least a portion of the reaction container is formed of a light-transmitting material; and The optical imaging sensor according to any one of claims 1 to 15, wherein the window portion of the optical imaging sensor is configured to be abutted against a portion of the reaction container formed of a light-transmitting material to monitor a biological reaction in the reaction container.

17. The bioreactor system according to claim 16, further comprising: A memory, wherein instructions are stored in the memory; as well as A processor is communicatively connected to the optical imaging sensor and the memory, and when the instructions are executed by the processor, image processing of the imaging image generated by the optical imaging sensor is implemented.

Images