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 comprising: 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 in a second direction, where the first direction and the second direction intersect with each other; the shell is configured to contain the illumination assembly and the detection assembly, the shell comprises a window part, and the window part is located on an emergent light path of the illumination assembly and located on an incident light path of the detection assembly, so that the illumination light beam and the imaging light beam can transmit and pass through the window part.

Description

Technical Field

[0001] 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

[0002] In the research and production of biological related fields, it is often necessary to monitor biological reaction processes such as cell culture. However, the current monitoring method is usually off-site and offline, that is, samples are extracted from the reaction container and then analyzed. This monitoring method is difficult to reflect the true state of the biological reaction in real time, and it is also impossible to provide continuous control signals to help achieve automatic control, and contamination may be introduced during the sampling process. Therefore, there is a need to improve the monitoring of biological reactions. Summary of the invention

[0003] One of the objectives of the present disclosure is to provide an optical imaging sensor and a biological response system.

[0004] According to a first aspect of the present disclosure, there is provided an optical imaging sensor, comprising:

[0005] an illumination assembly configured to generate an illumination beam along a first direction;

[0006] 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

[0007] 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.

[0008] In some embodiments, the lighting assembly comprises:

[0009] An illumination light source is configured to generate an initial light beam.

[0010] In some embodiments, the lighting assembly further comprises:

[0011] 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.

[0012] In some embodiments, the beam shaping assembly comprises:

[0013] 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

[0014] A cylindrical lens group, wherein the cylindrical lens group is arranged on the exit light path of the aperture;

[0015] 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.

[0016] 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 stop.

[0017] 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 component.

[0018] 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 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.

[0019] In some embodiments, the iris comprises an adjustable iris, and the light transmission range of the light-transmitting portion of the adjustable iris can be changed.

[0020] In some embodiments, the lighting assembly further comprises:

[0021] 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.

[0022] In some embodiments, the detection component includes:

[0023] an imaging lens group, the imaging lens group being configured to receive the imaging light beam; and

[0024] An optical detector is disposed on an outgoing light path of the imaging lens group and is configured to sense the imaging light beam.

[0025] In some embodiments, the imaging lens group includes:

[0026] objective lens; and

[0027] A tube lens is arranged between the objective lens and the optical detector.

[0028] In some embodiments, the detection component further comprises:

[0029] The filter is arranged on the incident light path of the optical detector, and the filter is configured to filter out the part of the imaging light beam in a preset wavelength band and allow the part of the imaging light beam in other wavelength bands to pass.

[0030] In some embodiments, the illumination assembly is positioned such that the illumination beam covers at least a focal plane of the detection assembly.

[0031] In some embodiments, the window portion is formed of a light-transmitting hard material or a light-transmitting flexible material.

[0032] In some embodiments, other portions of the housing except the window portion are formed of a non-light-transmitting material.

[0033] According to a second aspect of the present disclosure, there is provided a bioreactor system comprising:

[0034] A reaction container, wherein at least a portion of the reaction container is formed of a light-transmitting material; and

[0035] The optical imaging sensor as described above, wherein the window portion of the optical imaging sensor is configured to be attached to a portion of the reaction container formed of a light-transmitting material to monitor a biological reaction in the reaction container.

[0036] In some embodiments, the bioreactor system further comprises:

[0037] a memory having instructions stored therein; and

[0038] 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.

[0039] 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

[0040] The accompanying drawings, which constitute a part of the specification, illustrate embodiments of the present disclosure and, together with the description, serve to explain the principles of the present disclosure.

[0041] The present disclosure may be more clearly understood from the following detailed description with reference to the accompanying drawings, in which:

[0042] Figure 1 A schematic structural diagram of a bioreactor system according to a specific embodiment of the present disclosure is shown;

[0043] Figure 2 Shows Figure 1 Schematic diagram of the biological reaction system in working state;

[0044] Figure 3 Shows Figure 1 A partial enlarged schematic diagram of the biological reaction system in the working state;

[0045] Figure 4 A schematic structural diagram of a bioreactor system according to another specific embodiment of the present disclosure is shown;

[0046] Figure 5 A schematic structural diagram of an optical imaging sensor according to an exemplary embodiment of the present disclosure is shown;

[0047] Figure 6 A schematic structural diagram of an aperture according to a specific embodiment of the present disclosure is shown;

[0048] Figure 7 A schematic diagram showing an illumination light beam generated when a cylindrical lens group including a first cylindrical lens is used according to a specific embodiment of the present disclosure;

[0049] Figure 8 Shows Figure 7 Schematic diagram of the cross section of the illumination beam in the xz plane;

[0050] Fig. 9 Shows Figure 7 Schematic diagram of the cross section of the illumination beam in the xy plane;

[0051] Fig.10An imaging view of cells in a biological reaction system according to a specific application example of the present disclosure is shown;

[0052] Fig.11 A curve diagram showing the change of total cell density over time in a bioreactor system according to a specific application example of the present disclosure is shown;

[0053] Fig.12 A curve diagram showing the change of the density of living cells over time in a bioreactor system according to a specific application example of the present disclosure is shown;

[0054] Fig.13 A curve diagram showing changes in cell viability over time in a bioreactor system according to a specific application example of the present disclosure is shown;

[0055] Fig.14 A curve chart showing changes in cell diameter over time in a bioreactor system according to a specific application example of the present disclosure is shown.

[0056] Note that in the embodiments described below, the same reference numerals are sometimes used in common between different drawings to represent the same parts or parts with the same functions, and their repeated descriptions are omitted. In this specification, similar reference numerals and letters are used to represent similar items, so once an item is defined in one drawing, it does not need to be further discussed in subsequent drawings.

[0057] For ease of understanding, the positions, sizes, and ranges of various structures shown in the drawings and the like sometimes do not represent actual positions, sizes, and ranges, etc. Therefore, the disclosed invention is not limited to the positions, sizes, and ranges, etc. disclosed in the drawings and the like. In addition, the drawings are not necessarily drawn to scale, and some features may be exaggerated to show details of specific components. DETAILED DESCRIPTION

[0058] Various exemplary embodiments of the present disclosure will now be described in detail with reference to the accompanying drawings. It should be noted that 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 unless otherwise specifically stated.

[0059] 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 and its application or use. That is, the devices and methods herein are shown in an exemplary manner to illustrate different embodiments of the present disclosure and are not intended to be limiting. Those skilled in the art will appreciate that they merely illustrate exemplary ways that can be used to implement the present invention, rather than an exhaustive way.

[0060] Technologies, methods, and apparatus known to ordinary technicians in the relevant field may not be discussed in detail, but where appropriate, such technologies, methods, and apparatus should be considered part of the authorization specification.

[0061] In the research and production of biological related fields, the monitoring of reaction processes such as biological culture is often involved. At present, reaction vessels including culture dishes, shake flasks, reactors, disposable biological culture bags (e.g., cell culture bags) can be used to achieve the cultivation of cells or microorganisms. In particular, disposable biological culture bags are increasingly favored in large-scale cultivation such as biopharmaceuticals and synthetic biology.

[0062] 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, extracting samples from a reaction container (e.g., a reaction tank or a culture bag), and then performing cell analysis on the extracted samples. For example, a cell sensor based on dielectric spectrum or capacitance can be used to indirectly measure some parameters. In this measurement method, the electrical signal generated by the radio frequency electric field polarized cells can be collected in advance, and the correlation between the electrical signal and parameters such as the density of living cells can be established. During the measurement process, an offline detection device can be used to obtain the electrical signal measurement value, and the relevant parameters of the sample can be obtained according to the pre-established correlation. 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, the measurement accuracy is low, and the total number of cells (including living cells and dead cells) or total cell density and other parameters cannot be measured. 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.

[0063] 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. Further, it can be combined with, for example, image processing software to realize automatic 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 vessels 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 (e.g., cytokine secretion), cell characteristics (e.g., surface markers or genetic material), cell growth rate, and cell distribution at different levels in the reaction container.

[0064] In an exemplary embodiment of the present disclosure, the optical imaging sensor for imaging the sample to be tested may be provided separately, or may be integrated with a reaction vessel, for example, for culturing cells, to form at least a part of a biological reaction system. Figures 1 to 4 As shown, in some embodiments of the present disclosure, the biological reaction system may include a reaction vessel 200 and an optical imaging sensor 100. Among them, a stirring portion 210 and the like may also be provided in the reaction vessel 200, so as to stir the liquid containing the biological particles 900 therein during the reaction process to achieve a better reaction effect. At least a portion of the reaction vessel 200 may be formed of a light-transmitting material, so that the window portion 131 of the optical imaging sensor 100 can be abutted against the light-transmitting portion of the reaction vessel 200, thereby allowing the illumination light beam generated by the optical imaging sensor 100 to irradiate the sample in the reaction vessel 200, and the imaging light beam generated by the interaction between the illumination light beam and the biological particles 900 in the sample can pass through the reaction vessel 200 and return to the optical imaging sensor 100 for imaging. In a specific example, the entire reaction vessel 200 may be formed of a light-transmitting material, so that the window portion 131 of the optical imaging sensor 100 can be abutted against most positions on the reaction vessel 200 to monitor the biological reaction at the desired position. In other specific examples, only a portion of the reaction container 200 is formed of a light-transmitting material, while the other portion can be formed of an opaque material, so as to reduce unnecessary ambient light from entering the reaction container 200 as much as possible while ensuring the monitoring effect of the optical imaging sensor 100, thereby reducing the interference caused to the biological reaction therein. For example, the light-transmitting portion of the reaction container 200 can be a long strip extending along its longitudinal axis, so that the window portion 131 of the optical imaging sensor 100 can move along the longitudinal axis of the reaction container 200 to achieve monitoring of biological reactions at multiple levels in the reaction container 200.

[0065] exist Figures 1 to 3 In the specific example shown, when the biological reaction system is in operation, the reaction vessel 200 can be placed horizontally, the optical imaging sensor 100 can be arranged at the bottom of the reaction vessel 200, and the window portion 131 of the optical imaging sensor 100 can be attached to the light-transmitting portion of the reaction vessel 200. Figure 4 In the specific example shown, when the biological reaction system is in working state, the reaction vessel 200 can be placed vertically, and the optical imaging sensor 100 can be arranged on the side of the reaction vessel 200, for example, attached to the side wall of the reaction vessel 200, and the window portion 131 of the optical imaging sensor 100 can be attached to the light-transmitting portion of the reaction vessel 200.

[0066] In some embodiments, Figure 1 As shown, the bioreactor system may further include a memory 300 and a processor 400. For example, the memory 300 and the processor 400 may be part of a computer device, which is communicatively connected to the optical imaging sensor 100. In some embodiments, the computer device may be independently arranged, and 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, and when the instructions are executed by the processor 400, image processing of the imaging image generated by the optical imaging sensor 100 may be implemented, such as cell counting, viability analysis, etc. according to the imaging image. 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 result data of the image processing. In addition, in some embodiments, the memory 300 may also be directly communicatively connected to the optical imaging sensor 100, which is not limited here.

[0067] The memory 300 may be a volatile memory or a non-volatile memory, or may include both volatile and non-volatile memory. 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), synchronously 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.

[0068] The processor 400 may be an integrated circuit chip having signal processing capabilities. The processor may be a general-purpose processor, a digital signal processor (DSP), an application-specific integrated circuit (ASIC), an off-the-shelf 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 may be implemented or executed. The general-purpose processor may be a microprocessor or the processor may be any conventional processor, etc., and may be an X86 architecture or an ARM architecture, etc.

[0069] In an exemplary embodiment of the present disclosure, Figure 3 and Figure 5 As shown, the optical imaging sensor 100 may include an illumination component 110, a detection component 120, and a housing 130. The illumination component 110 may be configured to generate a light beam along a first direction (ie, Figure 5 The detection component 120 may be configured to receive and detect a second direction (ie, Figure 5 The first direction and the second direction may intersect or be perpendicular to each other; and the shell 130 may be configured to accommodate the lighting assembly 110 and the detection assembly 120, and the shell 130 may include a window portion 131, which may be located on an outgoing light path of the lighting assembly 110 and on an incident light path of the detection assembly 120, so that the lighting beam and the imaging beam can be transmitted through the window portion 131.

[0070] In some embodiments, Figure 5As shown, the lighting 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 sample to be tested may be observed in terms of morphology and the like based on the generated 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 convenient observation. Moreover, especially when the illumination light source 111 is an incoherent light source, it may generally have a smaller volume, and accordingly, the entire optical imaging sensor 100 may also have a smaller volume. For example, in the case of providing an incoherent light source, the characteristic dimension (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, so as to realize the fluorescence detection of the sample to be tested based on the generated fluorescence signal to obtain the relevant information of the biological particles 900. In this case, in order to improve the excitation effect, the illumination light beam can be a coherent light beam in the desired wavelength band, and accordingly, the illumination light source 111 can include a coherent light source such as a laser, or can include an optical fiber for transmitting a coherent light beam, etc. It can be understood that in some embodiments, the illumination light source 111 can also be a mixed light source that can generate both an incoherent light beam and a coherent light beam, or can include multiple illumination light sources 111 to switch to the corresponding illumination light beam as needed.

[0071] In some embodiments, the lighting assembly 110 may further include a beam shaping assembly, which may be disposed on the outgoing light path of the lighting light source 111, and the beam shaping assembly may be configured to shape the light beam to generate a lighting beam having a preset shape to achieve a better monitoring effect. Figure 5 As shown, the beam shaping assembly may include an aperture 112 and a cylindrical lens group 113, and the aperture 112 and the cylindrical lens group 113 may be configured to shape the light beam to generate a substantially sheet-shaped illumination beam, and the plane where the illumination beam is located may be perpendicular or substantially perpendicular to the second direction.

[0072] exist Figure 5 In the specific example shown, the aperture 112 may be disposed on the outgoing light path of the illumination light source 111. Figure 6 As shown, the aperture 112 may include a light shielding screen 112a and a light-transmitting portion 112b opened in the light shielding screen 112a, wherein the shape of the light-transmitting portion 112b may affect the illumination beam in the Figure 5The shape of the cross section on the yz plane (wherein the y direction is perpendicular to both the x direction and the z direction) shown in . In some embodiments, the light-transmitting portion 112b may be rectangular. In a specific example, the light-transmitting portion 112b may be square. In order to provide substantially uniform illumination in the observation field of the detection assembly 120 so as to better observe the sample to be tested and more conveniently calculate the required parameters, the effective width of the illumination light beam (the width of a portion of the light beam where the light intensity is above a certain threshold to provide sufficient illumination resolution) may be greater than the field of view width of the detection assembly 120 (or the objective lens 123 in the detection assembly 120 as described below), so that the illumination light beam may be distributed throughout the field of view. Further, in some embodiments, the diaphragm 112 may include an adjustable diaphragm, the light transmission range of the light-transmitting portion of the adjustable diaphragm being able to be changed so as to change the shape of the illumination light beam as needed, such as changing the effective width of the illumination light beam to achieve a desired illumination range.

[0073] exist Figure 5 In the specific example shown, the cylindrical lens group 113 can be arranged on the exit light path of the aperture 112. As described above, the cylindrical lens group 113 can shape the initial light beam into a sheet-shaped illumination light beam together with the aperture 112, and the normal direction of the sheet-shaped light beam is substantially along the second direction, and the plane of the sheet-shaped light beam extends in the xy plane. It should be noted that the sheet-shaped light beam described herein refers to a light beam whose thickness in the z direction is much smaller (for example, more than 10 times smaller) than its effective length in the x direction (the length of a portion of the light beam where the light intensity is above a certain threshold to provide sufficient illumination resolution), and the thickness of the light beam is much smaller than its effective width in the y direction.

[0074] The cylindrical lens group 113 can have a variety of settings to produce a desired sheet-shaped light beam. In addition, one or more cylindrical lenses used in the cylindrical lens group 113 can each be a single-piece cylindrical lens, or can be a multi-piece apochromatic cylindrical lens, so that the effective illumination part of the sheet-shaped light beam is more uniform and the influence of chromatic aberration on optical observation is avoided.

[0075] In a specific example, if Figure 5 As shown, the cylindrical lens group 113 may include a first cylindrical lens, wherein the convex surface of the first cylindrical lens may be arranged facing the aperture 112. In this case, the first cylindrical lens may generate a Figures 7 to 9 In the sheet-shaped beam shown in FIG. , the edge of the sheet-shaped beam extending along the x-direction may be bent to a certain extent.

[0076] 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 according to the diffraction limit. For example, in the case where 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 illumination beam 119 in the third direction (ie, Figure 5 The effective width in the direction of the y-axis in the middle) 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 light beam 119, and D is the wavelength of the light-transmitting portion 112b in the second direction (i.e., Figure 5 The side length of the light-transmitting portion 112b in the direction of the z-axis (i.e., Figure 5 , 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 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 can be The positions are determined to correspond to the two end points of the effective length of the illumination light beam.

[0077] In some 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, wherein 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 width of the light-transmitting portion 112b in the third direction (i.e., Figure 5 Similarly, the thickness of the illumination beam 119 can be The positions are determined to correspond to the two end points of the effective length of the illumination beam 119.

[0078] In some embodiments, in order to further reduce the thickness of the illumination beam 119, 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 beam more concentrated to better excite the fluorescent signal in the sample to be tested, such as Figure 5 As shown, the lighting assembly 110 may further include a collimating lens group 114, which may be disposed on the exit light path of the beam shaping assembly, and the collimating lens group 114 may be configured to collimate the light beam to generate a collimated lighting beam, for example, to make the thickness of the lighting 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.

[0079] Furthermore, if Figure 5 As shown, 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 come 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, and the tube lens 122 is disposed between the objective lens 123 and the optical detector 121. It is understood that in some other embodiments, the imaging lens group for collecting the imaging light beam may also be arranged in other ways, which is not limited here.

[0080] The optical detector 121 may be disposed on the outgoing light path of the imaging lens group, and the optical detector 121 may 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 fluorescent signal image). In a specific example, the optical detector 121 may include a charge coupled device (CCD) sensor or a complementary metal oxide semiconductor (CMOS) sensor. In addition, by making the light sheet scan 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, and then realizing three-dimensional information reconstruction of the sample to be tested.

[0081] In some embodiments, particularly in the case of fluorescence imaging, such as Figure 5As shown, the detection component 120 may also include a filter 124, which may be disposed on the incident light path of the optical detector 121, and the filter 124 may be configured to filter out the portion of the imaging light beam in a preset wavelength band, and allow the portion of the imaging light beam in other wavelength bands to pass (for example, filter out a portion of the imaging light beam in the wavelength band of the illumination light beam, and allow the fluorescence signal to pass). In this way, the interference caused by light outside the wavelength band of the fluorescence signal on the detection can be reduced, so as to help better detect the sample to be tested.

[0082] In some embodiments, in order to achieve better imaging effects, such as Figure 5 As shown, the position of the lighting assembly 110 can be set so that the lighting beam at least covers the focal plane of the detection assembly 120, so that the detection assembly 120 only senses the imaging of biological particles located near the focal plane of the imaging lens group. In other words, the lighting assembly 110 and the detection assembly 120 satisfy such a positional relationship, so that the area irradiated by the sheet-shaped lighting beam is basically located on the focal plane of the detection assembly 120, thereby forming the best imaging effect.

[0083] In some embodiments, Figure 3 and Figure 5 As shown, 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 biological reactions 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 according to the desired optical path. In some embodiments, the window portion 131 can be formed by a light-transmitting hard material (e.g., quartz). In this way, especially in the case of monitoring biological reactions in a reaction vessel 200 formed by a flexible material, the hard window portion 131 is conducive to making the area of ​​the reaction vessel 200 against the optical imaging sensor 100 more flat, thereby improving the observation effect. In other embodiments, the window portion 131 can also be formed by a light-transmitting flexible material (e.g., polyethylene (PE)). In this way, the window portion 131 can conformally abut against the surface of the reaction container 200 of various shapes to improve the monitoring effect. In addition, in some embodiments, the other parts of the housing 130 except 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 a plurality of independent parts, or the housing 130 can also be formed in one piece, which is not limited here.

[0084] Some specific application examples in optical imaging sensors and bio-response systems are listed below.

[0085] 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 feed (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 did not stop growing, but instead grew in a different way, that is, the increase in cell number was changed to an increase in single-cell biomass, at which time the cell volume and diameter would 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, Fig.10 Figure 1 shows an optical imaging photograph of cells 900 obtained during the culture process. Fig.10 The optical imaging photos shown can be used to extract or calculate the changes in the total cell density (TCD) of CHO cells over time using relevant image processing software (such as Fig.11 As shown, the horizontal axis represents days, and the vertical axis represents TCD and the change of viable cell density (VCD) over time (as shown in Fig.12 As shown, the horizontal axis represents days, the vertical axis represents VCD, the cell viability (Via) changes over time (such as Fig.13 As shown, the horizontal axis represents the number of days, and the vertical axis represents the changes of Via) and diameter (Dia) over time (as shown in Fig.14 As shown, the horizontal axis represents days, the vertical axis represents Dia), and other information, and the cell growth curve can be further drawn, etc., to provide timely and effective assistance to scientific researchers in process optimization and changes in antibody production.

[0086] 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, and when the depth of field of the objective lens 123 is greater than the thickness Z of the illumination beam, the effective volume is obtained according to the product of the thickness Z of the illumination beam and the field of view area, and the cell density is calculated based on the effective volume. Alternatively, when 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 according to the product of the depth of field of the objective lens 123 and the field of view area, and the cell density in the sample to be tested is calculated based on the effective volume.

[0087] 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) of the detection component is imaged, and by detecting the imaging in the orthogonal direction, the influence of the out-of-focus background on the imaging quality can be effectively avoided, and at the same time, a high imaging contrast can be achieved and optical tomography can be naturally achieved to construct a three-dimensional model, which greatly reduces the photobleaching and phototoxicity of the illumination light beam to the sample to be tested, and is therefore suitable for long-term imaging of living biological samples for hours or even days, without the 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 container, the detection and statistics of parameters such as the counting, density, viability, volume, grouping classification, and growth rate of biological samples can be achieved. In addition, the optical imaging sensor disclosed in the present invention can be integrated into a housing, which 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 part 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 pollute 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 preliminary treatment is saved, and the influence of the chemical toxicity of the reagent itself on the cell state and characteristic expression is avoided, thereby helping to achieve in-situ, real-time, direct and continuous cell dynamic analysis.

[0088] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limiting. Therefore, other examples of the exemplary embodiments may have different values.

[0089] The words "front", "rear", "top", "bottom", "above", "below", etc., if present, in the specification and claims are used for descriptive purposes and are not necessarily used to describe invariant relative positions. It should be understood that the words 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.

[0090] As used herein, the word "exemplary" means "serving as an example, instance, or illustration," rather than as a "model" to be exactly copied. Any implementation described as an example herein is not necessarily to be construed as preferred or advantageous over other implementations. Furthermore, the present disclosure is not limited by any expressed or implied theory given in the above technical field, background technology, summary of the invention, or detailed description.

[0091] As used herein, the term "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 term "substantially" also allows for deviations from a perfect or ideal condition due to parasitic effects, noise, and other practical considerations that may exist in actual implementations.

[0092] The above description may indicate an element or node or feature that is "connected" or "coupled" together. As used herein, unless otherwise expressly stated, "connection" means that one element / node / feature is directly connected (or directly communicates) to another element / node / feature electrically, mechanically, logically or otherwise. Similarly, unless otherwise expressly stated, "coupling" 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, "coupling" is intended to include direct and indirect connections of elements or other features, including connections using one or more intermediate elements.

[0093] 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.

[0094] Those skilled in the art will appreciate that the boundaries between the above operations are merely illustrative. Multiple operations can be combined into a single operation, a single operation can be distributed in additional operations, and operations can be performed at least partially overlapping in time. Moreover, alternative embodiments may include multiple instances of specific operations, and the order of operations may be changed in other various embodiments. However, other modifications, variations, and replacements are equally possible. Therefore, this specification and accompanying drawings should be considered illustrative, not restrictive.

[0095] Although some specific embodiments of the present disclosure have been described in detail by way of example, 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, characterized in that: The optical imaging sensor comprises: 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; and 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: 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, characterized in that: The detection component also includes: The filter is arranged on the incident light path of the optical detector, and the filter is configured to filter out the part of the imaging light beam in a preset wavelength band and allow the part of the imaging light beam in other wavelength bands to pass.

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

16. A bioreactor system, characterized in that: The bioreactor system comprises: 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, characterized in that: The bioreactor system further comprises: a memory having instructions stored therein; and 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.

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