Optical imaging sensors and particle analysis equipment

Abstract

The present disclosure relates to an optical imaging sensor and a particle analysis device, wherein the optical imaging sensor includes an aperture assembly arranged on an imaging light path, the aperture assembly including: a first aperture, the first aperture having a first light-shielding portion and a first light-transmitting portion, the first light-transmitting portion being configured to allow at least a portion of an imaging light beam located in a first area to pass through when participating in imaging, so as to form a first image; and a second aperture, the second aperture having a second light-shielding portion and a second light-transmitting portion, the second light-transmitting portion being configured to allow at least a portion of an imaging light beam located in a second area different from the first area to pass through when participating in imaging, so as to form a second image, wherein the relative position of the second light-transmitting portion on the second aperture is different from the relative position of the first light-transmitting portion on the first aperture.

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 particle analysis device. Background Art

[0002] Particle analysis usually includes the analysis and statistics of relevant information such as particle counting, shape (round or quasi-circular), surface morphology (smooth or rough), skeleton structure (porous or solid). Common particle analysis methods include analysis based on the Coulter principle, which is mainly used for cell counting and cell volume detection. In addition, particle analysis methods can also include flow cytometry, which targets single cells or other biological particles in suspension, and detects the fluorescent signal of the marker to achieve high-speed, one-by-one cell quantitative analysis and sorting technology. It is widely used to analyze the expression of cell surface and intracellular molecules, identify and determine different cell types in heterogeneous cell populations, evaluate the purity of separated subpopulations, and analyze cell size and volume. In addition, particle analysis methods can also include optical microscopy combined with staining and labeling observation. However, the above-mentioned particle analysis methods each have their corresponding problems, so it is necessary to improve the method of particle analysis. Summary of the invention

[0003] One of the objectives of the present disclosure is to provide an optical imaging sensor and a particle analysis device.

[0004] According to a first aspect of the present disclosure, an optical imaging sensor is provided, comprising an aperture assembly arranged on an imaging optical path, the aperture assembly comprising:

[0005] a first aperture, the first aperture having a first light shielding portion and a first light transmitting portion, the first light transmitting portion being configured to transmit at least a portion of the imaging light beam located in the first region when participating in imaging, so as to form a first image; and

[0006] A second aperture, the second aperture having a second light-shielding portion and a second light-transmitting portion, the second light-transmitting portion being configured to allow at least a portion of an imaging light beam located in a second area different from the first area to pass through when participating in imaging, so as to form a second image, wherein the relative position of the second light-transmitting portion on the second aperture is different from the relative position of the first light-transmitting portion on the first aperture.

[0007] In some embodiments, the first light shielding portion is arranged to surround the first light transparent portion; and

[0008] The second light-transmitting portion is arranged to surround the second light-shielding portion.

[0009] In some embodiments, the relative position of the first light-transmitting portion on the first aperture is equivalent to the relative position of the second light-blocking portion on the second aperture; and

[0010] The relative position of the first light-blocking portion on the first aperture is equivalent to the relative position of the second light-transmitting portion on the second aperture.

[0011] In some embodiments, the optical imaging sensor further includes:

[0012] A condenser lens, which is disposed on the incident light path and is configured to converge the incident light beam; and

[0013] An objective lens, which is disposed on the imaging light path and is configured to collect the imaging light beam.

[0014] In some embodiments, NAc < NAo, where NAc is the numerical aperture of the condenser lens and NAo is the numerical aperture of the objective lens.

[0015] In some embodiments, when participating in imaging, the first aperture is located at the rear focal plane of the objective lens, the first light-transmitting portion is circular and located in the central region of the first aperture, and the radius r11 of the first light-transmitting portion satisfies r11 = NAc * f; and / or

[0016] When participating in imaging, the second aperture is located at the rear focal plane of the objective lens, the second light-blocking portion is circular and located in the central region of the second aperture, and the radius r22 of the second light-blocking portion satisfies r22 = NAc * f;

[0017] where f is the focal length of the objective lens.

[0018] In some embodiments, when participating in imaging, the first aperture is located between the rear focal plane and the exit pupil plane of the objective lens, the first light-transmitting portion is circular and located in the central region of the first aperture, and the radius r11 of the first light-transmitting portion satisfies r11 = NAc * (f - s1); and / or

[0019] When participating in imaging, the second aperture is located between the rear focal plane and the exit pupil plane of the objective lens, the second light-blocking portion is circular and located in the central region of the second aperture, and the radius r22 of the second light-blocking portion satisfies r22 = NAc * (f - s2);

[0020] where f is the focal length of the objective lens, s1 is the distance between the first aperture and the objective lens when participating in imaging, and s2 is the distance between the second aperture and the objective lens when participating in imaging.

[0021] In some embodiments, the optical imaging sensor further comprises:

[0022] A switching member is mechanically connected to the first aperture and the second aperture, and is configured to switch one of the first aperture and the second aperture to be located on the imaging optical path, and to switch the other of the first aperture and the second aperture to be away from the imaging optical path.

[0023] In some embodiments, the optical imaging sensor further comprises:

[0024] An imaging mirror, wherein the imaging mirror is located on an exit light path of the objective lens, and the aperture assembly is located between the objective lens and the imaging mirror; and

[0025] A detector is located on the output light path of the imaging mirror, and the detector is configured to perform imaging based on a portion of the imaging light beam.

[0026] In some embodiments, the optical imaging sensor further comprises:

[0027] a beam splitter, the beam splitter being located on an exit optical path of the objective lens and being configured to allow a portion of the imaging light beam to travel along a first optical path and another portion of the imaging light beam to travel along a second optical path;

[0028] The first optical path and the second optical path intersect each other, the first aperture is located on the first optical path, and the second aperture is located on the second optical path.

[0029] In some embodiments, the optical imaging sensor further comprises:

[0030] A first imaging mirror, wherein the first imaging mirror is located on an exit light path of the first aperture;

[0031] a first detector, the first detector being located on an exit light path of the first imaging mirror, and the first detector being configured to perform imaging based on a portion of the imaging light beam;

[0032] A second imaging mirror, the second imaging mirror is located on the exit light path of the second aperture; and

[0033] A second detector is located on the output light path of the second imaging mirror, and the second detector is configured to perform imaging based on a portion of the imaging light beam.

[0034] In some embodiments, the optical imaging sensor further includes a light source configured to generate an incident light beam.

[0035] According to a second aspect of the present disclosure, there is provided a particle analysis device, comprising:

[0036] flow cell; and

[0037] The optical imaging sensor as described above, wherein the optical imaging sensor is configured to generate a first image and a second image based on the sample in the flow cell.

[0038] In some embodiments, the particle analysis device further comprises:

[0039] Memory; and

[0040] A processor is configured to analyze imaging data from the optical imaging sensor to generate target data.

[0041] 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

[0042] 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.

[0043] The present disclosure may be more clearly understood from the following detailed description with reference to the accompanying drawings, in which:

[0044] Figure 1 A schematic structural diagram of an optical imaging sensor according to an exemplary embodiment of the present disclosure is shown;

[0045] Figure 2 A schematic structural diagram of an optical imaging sensor according to another exemplary embodiment of the present disclosure is shown;

[0046] Figure 3 A block diagram of a particle analysis device according to an exemplary embodiment of the present disclosure is shown;

[0047] FIG4( a ) shows a first image generated in an optical imaging sensor according to a specific example of the present disclosure;

[0048] FIG4( b ) shows a second image generated in an optical imaging sensor according to a specific example of the present disclosure;

[0049] Figure 5 A schematic diagram showing changes in live cell density and cell viability over time during a cell culture process according to another specific example of the present disclosure is shown;

[0050] FIG6( a ) shows a first image generated in an optical imaging sensor according to another specific example of the present disclosure;

[0051] FIG6( b ) shows a second image generated in an optical imaging sensor according to another specific example of the present disclosure;

[0052] Figure 7 A schematic diagram of cell clustering during a cell culture process according to another specific example of the present disclosure is shown.

[0053] 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.

[0054] 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

[0055] 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.

[0056] 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 optical imaging sensor and particle analysis device 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 understand that they merely illustrate exemplary ways that can be used to implement the present invention, rather than exhaustive ways.

[0057] 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.

[0058] In general, particles such as cells can be analyzed based on the Coulter principle, flow cytometry or optical microscopy combined with staining and labeling observation, including particle counting, shape (round or quasi-circular), surface morphology (smooth or rough), skeleton structure (porous or solid), etc. Among them, the analysis based on the Coulter principle is indirect, and usually can only roughly obtain information related to the number and density of living cells. The measurement accuracy is low, and it is difficult to continuously and in real time monitor the cell growth state. At the same time, the process of extracting samples may also introduce the risk of cross-contamination; flow cytometry is good at statistics, but lacks imaging capabilities, and cannot obtain information such as cell morphology and subcellular localization. In addition, the sample pretreatment of flow cytometry is extremely complex and time-consuming, requiring operations such as centrifugation, pipetting, mixing, gradient dilution, and light-proof incubation of sample reagents; while optical microscope imaging technology is good at morphological analysis and can provide detailed particle image information, but requires pre-staining and other treatments. It can be seen that the above-mentioned particle analysis methods have their own problems.

[0059] In order to solve the above problems, the present disclosure proposes an optical imaging sensor and a particle analysis device, which can detect and analyze information such as particle morphology, diameter, particle size, and transmittance based on the combination of video imaging and image processing, without the need for sample pretreatment such as staining.

[0060] In an exemplary embodiment of the present disclosure, Figure 1 and Figure 2 As shown, the optical imaging sensor may include an aperture assembly disposed on an imaging optical path, and an imaging light beam from the sample 900 may be irradiated onto a corresponding detector via the imaging optical path to generate an imaging image or an imaging video for particle analysis. Figure 1 and Figure 2 As shown, the aperture assembly may include a first aperture 141 and a second aperture 142. It should be noted that Figure 1 and Figure 2 In the figure, for the sake of clarity, the main planes of the first aperture 141 and the second aperture 142 are shown. In actual applications, the main planes of the first aperture 141 and the second aperture 142 will be set to be perpendicular to the optical axis, that is, parallel to the plane perpendicular to the paper surface, so as to filter out the desired part from the imaging light beam from the sample 900 for imaging.

[0061] The first aperture 141 may have a first light shielding portion (such as Figure 1 and Figure 2 The first aperture 141 is shown in black) and the first light-transmitting portion (as shown in FIG. Figure 1 and Figure 2The first light-transmitting portion may be configured to transmit at least a portion of the imaging light beam located in the first region when participating in the imaging to form a first image. Figure 1 and Figure 2 In the specific example shown, the first light shielding portion can be arranged to surround the first light transmitting portion. That is, when the first aperture 141 is in the optical path and thus participates in imaging, the first aperture 141 can allow the central portion of the imaging light beam to pass through, while blocking the edge portion of the imaging light beam. The first imaging generated based on the central portion of the light beam mainly corresponds to information such as the morphology and diameter of the particles, that is, the first imaging is mainly formed by the transmission (low-frequency) signal, as shown in FIG. 4( a) and FIG. 6( a), which can be used to analyze and count the number and shape of the particles.

[0062] The second aperture 142 may have a second light shielding portion (such as Figure 1 and Figure 2 The second aperture 142 is shown in black) and the second light-transmitting portion (as shown in FIG. Figure 1 and Figure 2 The second light-transmitting portion may be configured to transmit at least a portion of the imaging light beam located in a second region different from the first region when participating in the imaging, so as to form a second image. Figure 1 and Figure 2 As shown, the relative position of the second light-transmitting portion on the second aperture 142 may be different from the relative position of the first light-transmitting portion on the first aperture 141. For example, the second light-transmitting portion may be arranged to surround the second light-shielding portion. That is, when the second aperture 142 is in the optical path and thus participates in imaging, the second aperture 142 may allow the edge portion of the imaging light beam to pass through, while blocking the center portion of the imaging light beam. The second imaging generated based on the edge portion of the light beam mainly corresponds to information such as the particle size and transmittance of the particles, that is, the second imaging is mainly formed by the scattered (high-frequency) signal, as shown in FIG4(b) and FIG6(b), which can be used to analyze and count the surface morphology and skeleton structure of the particles.

[0063] Based on the above description, it can be known that by setting the first aperture 141 and the second aperture 142, different parts of the imaging light beam can be screened out for imaging, thereby forming different imaging effects to better achieve analysis and statistics of relevant properties of particles. It can be understood that in some other embodiments, as needed, the first aperture 141 and the second aperture 142 can also have other settings, not limited to Figure 1 and Figure 2As shown in , as long as the relative positions of the light-transmitting parts of the first aperture 141 and the second aperture 142 are different, different parts of the imaging light beam can be screened out for targeted imaging. In addition, in some embodiments, the aperture assembly can also include more apertures, and the light-transmitting parts of different apertures can have different relative positions in the aperture to respectively form desired imaging effects, which is not limited here.

[0064] In some embodiments, Figure 1 and Figure 2 As shown, the relative position of the first light-transmitting portion on the first aperture 141 may be equivalent to the relative position of the second light-shielding portion on the second aperture 142, and the relative position of the first light-shielding portion on the first aperture 141 may be equivalent to the relative position of the second light-transmitting portion on the second aperture 142. In other words, the first aperture 141 and the second aperture 142 may be complementary to each other, and the portion of the imaging light beam that passes through the first aperture 141 and the portion that passes through the second aperture 142 may be combined into a complete imaging light beam, so that such an optical imaging sensor can be used to analyze each portion of the entire imaging light beam separately without missing a certain portion of the imaging light beam. However, in some other embodiments, the first aperture 141 and the second aperture 142 may not be complementary. For example, the relative position of the first light-shielding portion of the first aperture 141 and the relative position of the second light-shielding portion of the second aperture 142 may overlap to a certain extent, so that a portion of the imaging light beam may not participate in any imaging, thereby effectively removing the influence of the undesirable portion of the imaging light beam on the final imaging effect. Alternatively, the relative position of the first light-transmitting portion of the first aperture 141 and the relative position of the second light-transmitting portion of the second aperture 142 may overlap to a certain extent, that is, part of the imaging light beam may be used to form both the first image and the second image to produce the desired imaging effect, which is not limited here.

[0065] In an exemplary embodiment of the present disclosure, Figure 1 and Figure 2 As shown, the optical imaging sensor may further include a light source 110. The light source 110 may be configured to generate an incident light beam. The incident light beam may be converged by a condenser lens 120 as described below to illuminate the sample 900 for imaging.

[0066] In some embodiments, the light source 110 may be configured to generate incoherent light such as white light, for example, the light source 110 may be formed by a light emitting device such as a light emitting diode (LED). Alternatively, if necessary, the light source 110 may also be configured to generate coherent light having a specific wavelength, for example, the light source 110 may be a laser, etc., which is not limited here.

[0067] like Figure 1 and Figure 2As shown, in some embodiments, the optical imaging sensor may further include a condenser lens 120. The condenser lens 120 may be composed of one or more lenses. The condenser lens 120 may be disposed on the incident light path, or located at the incident end of the sample 900, and the condenser lens 120 may be configured to converge the incident light beam to form a better imaging effect.

[0068] In some embodiments, as Figure 1 and Figure 2 shown, the optical imaging sensor may further include an objective lens 130. The objective lens 130 may be disposed on the imaging light path, or located at the exit end of the sample 900, and the objective lens 130 may be configured to collect the imaging light beam. Here, the objective lens 130 may be composed of one or more lenses, which is not limited herein.

[0069] In some embodiments, in the optical imaging sensor, the following condition is satisfied: NAc < NAo, where NAc is the numerical aperture of the condenser lens 120 and NAo is the numerical aperture of the objective lens 130.

[0070] Further, in some embodiments, when the first aperture 141 participates in imaging, it may be located at the rear focal plane of the objective lens 130. When NAc < NAo and the first light-transmitting portion is a circle located in the central region of the first aperture 141, the radius r11 of the first light-transmitting portion may satisfy r11 = NAc * f to better separate the low-frequency portion in the imaging light beam, where f is the focal length of the objective lens 130. Similarly, when the second aperture 142 participates in imaging, it may also be located at the rear focal plane of the objective lens 130. When NAc < NAo and the second light-blocking portion is a circle located in the central region of the second aperture 142, the radius r22 of the second light-blocking portion may satisfy r22 = NAc * f to better block the low-frequency portion in the imaging light beam, and allow the high-frequency portion in the imaging light beam to pass through the second light-transmitting portion around the second light-blocking portion for imaging.

[0071] In some other embodiments, the first aperture 141 or the second aperture 142 may not be disposed on the rear focal plane of the objective lens 130 either. For example, when the first aperture 141 participates in imaging, it may also be located between the rear focal plane and the exit pupil plane of the objective lens 130. When NAc < NAo and the first light-transmitting portion is circular and located in the central region of the first aperture 141, the radius r11 of the first light-transmitting portion may satisfy r11 = NAc * (f - s1) to better separate the low-frequency portion in the imaging beam, where f is the focal length of the objective lens 130 and s1 is the distance between the first aperture 141 and the objective lens 130 when the first aperture 141 participates in imaging. Similarly, when the second aperture 142 participates in imaging, it may be located between the rear focal plane and the exit pupil plane of the objective lens 130. When NAc < NAo and the second light-blocking portion is circular and located in the central region of the second aperture 142, the radius r22 of the second light-blocking portion may satisfy r22 = NAc * (f - s2) to better block the low-frequency portion in the imaging beam and allow the high-frequency portion in the imaging beam to pass through the second light-transmitting portion around the second light-blocking portion for imaging, where s2 is the distance between the second aperture 142 and the objective lens 130 when the second aperture 142 participates in imaging.

[0072] In each imaging process, only one aperture in the aperture assembly participates in imaging. To achieve this effect, various methods can be used to switch between different apertures. For example, in a specific example, the optical imaging sensor 100 may further include a switching member, which can be mechanically connected to the first aperture 141 and the second aperture 142, and the switching member can be configured to switch one of the first aperture 141 and the second aperture 142 to be located on the imaging optical path and switch the other of the first aperture 141 and the second aperture 142 to be off the imaging optical path, so that only one aperture is located on the imaging optical path to screen out the desired portion in the imaging beam to produce the desired imaging effect. The switching member may include structures such as a turntable, a connecting rod, a gear, etc., which are not limited herein. For example, multiple apertures can be disposed on the same turntable, and by rotating the turntable, the desired aperture can be set in the imaging optical path to participate in imaging. Alternatively, structures such as a connecting rod and a gear can also be used to send the desired aperture into the imaging optical path and remove other apertures.

[0073] In Figure 1In the specific example shown, a switching member can be used to select the desired aperture. In addition, the optical imaging sensor can also include an imaging lens 150 and a detector 160, and the imaging lens 150 can be located on the exit light path of the objective lens 130, and the aperture assembly can be located between the objective lens 130 and the imaging lens 150. Through the switching member, the desired aperture can be set on the objective lens 130 and the imaging lens 150 to participate in imaging. In a specific example, the imaging lens 150 can be formed by a tube lens, which can include one or more lenses to converge parallel light beams from an infinite objective lens into an image. The detector 160 can be located on the exit light path of the imaging lens 150, and the detector 160 can be configured to perform imaging based on a portion of the imaging light beam, which is the portion of the light beam filtered out by the aperture in the imaging light path. Here, the detector 160 can be formed by a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS) image sensor, etc. Since the switching of different apertures can be achieved based on the switching component, multiple apertures in the aperture assembly can share the same set of objective lens 130, imaging lens 150 and detector 160, so as to help reduce the cost of the optical imaging sensor.

[0074] In another specific example, a corresponding imaging optical path may be set for each aperture to achieve imaging. Figure 2 As shown, the optical imaging sensor may further include a beam splitter 170, which may be located on the exit optical path of the objective lens 130, and the beam splitter 170 may be configured to cause a portion of the imaging light beam to travel along a first optical path and another portion of the imaging light beam to travel along a second optical path, wherein the first optical path and the second optical path intersect with each other, the first aperture 141 may be located on the first optical path, and the second aperture 142 may be located on the second optical path.

[0075] Furthermore, if Figure 2As shown, the optical imaging sensor may further include a first imaging mirror 151, a first detector 161, a second imaging mirror 152, and a second detector 162, which are arranged corresponding to each aperture. The first imaging mirror 151 may be located on the exit optical path of the first aperture 141. Similarly, the first imaging mirror 151 may be formed by a tube lens, which may include one or more lenses to converge the parallel light beams from the infinite objective lens into an image. The first detector 161 may be located on the exit optical path of the first imaging mirror 151, and the first detector 161 may be configured to perform imaging based on a portion of the imaging light beam. Here, the first detector 161 may be formed by a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS) image sensor, etc. Similarly, the second imaging mirror 152 may be located on the exit optical path of the second aperture 142, which may include one or more lenses to converge the parallel light beams from the infinite objective lens into an image. In addition, the second detector 162 may be located on the exit optical path of the second imaging mirror 152, and the second detector 162 may be configured to perform imaging based on a portion of the imaging light beam. Here, the second detector 162 may also be formed by a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS) image sensor, etc. By respectively providing corresponding imaging mirrors and detectors for each aperture for imaging, the adverse effects or interference of switching on the optical path can be reduced, the stability of the optical path is improved, and the optical path of each aperture can be optimized, thereby improving the imaging effect.

[0076] It is understandable that in some other embodiments, other optical devices such as lenses and reflectors may be provided in the optical imaging sensor as required, which is not limited here.

[0077] The present disclosure also proposes a particle analysis device, such as Figure 3 As shown, the particle analysis device may include a circulation cell 400 and the optical imaging sensor 100 as described above, wherein the optical imaging sensor 100 may be configured to generate a first image and a second image based on the sample in the circulation cell 400. In some embodiments, a plurality of first images acquired over time may be formed into a first video, and similarly, a plurality of second images acquired over time may be formed into a second video for further analysis.

[0078] Furthermore, in some embodiments, Figure 3 As shown, the particle analysis device may further include a memory 200 and a processor 300 .

[0079] The memory 200 may be a volatile memory or a nonvolatile memory, or may include both volatile and nonvolatile memories. The nonvolatile 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 connection dynamic random access memory (SLDRAM), and direct memory bus random access memory (DR RAM). It should be noted that the memory of the method described herein is intended to include, but is not limited to, these and any other suitable types of memory. The memory 200 may be used to store, for example, the first imaging and the second imaging, and may also be used to store intermediate data, result data, etc. generated during the processing of the first imaging and the second imaging.

[0080] The processor 300 can be configured to analyze the imaging data from the optical imaging sensor 100 to generate target data, including analysis or statistical results of particle counts, shapes, surface morphologies or skeleton structures. The processor 300 can also perform various actions and processes according to instructions stored in the memory 200. Specifically, the processor 300 can be an integrated circuit chip with signal processing capabilities. The above-mentioned processor 300 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 devices, discrete gates or transistor logic devices, discrete hardware components. The various methods, steps and logic block diagrams disclosed in the embodiments of the present disclosure can be implemented or executed. The general-purpose processor can be a microprocessor or the processor can also be any conventional processor, etc., which can be an X86 architecture or an ARM architecture, etc.

[0081] The following is a specific application example of optical imaging sensors and particle analysis equipment, in which the status and subtypes of CHO cells under long-term culture conditions are monitored. CHO cells are mainly used for antibody production in the biopharmaceutical field.

[0082] Generally, during the cell culture process, the growth curve of the culture can be divided into several different stages: 1) the initial lag phase, when cells need to adapt to the new environment and thus grow slowly; 2) the exponential growth phase (or logarithmic phase), once the cells adapt to the environment and continue to divide at a constant rate, they will enter the exponential growth phase or rapid growth phase; 3) the stable growth phase, once the nutrients are exhausted by the cells and byproducts accumulate, the cell growth begins to slow down and enter a stable state; 4) the death phase, after the stable phase, the density of live cells drops sharply and enters the death phase. Usually, the culture and products are harvested during the death phase.

[0083] For example, CHO cells can be cultured in a bioreactor system for IgG expression. Figure 5 As shown, the bioreactor inoculation density was 5x10 6 cells / mL, one day after inoculation, the cells were switched to perfusion mode. Within 8 days, the cell density increased to 80x10 6 cells / mL, the cell viability was about 90%. Throughout the process, the IgG screening coefficient was between 91% and 99%. 6 cells / mL, process control began to deteriorate, CO2 was not adequately removed from the reactor, and the dissolved oxygen demand of high-density culture could not be met. Attempts were made to increase the stirring speed and the bottom gas flow rate to 0.07 VVM to control DO above the set value, while a lot of foaming was generated. Cell viability remained at a high level, indicating that CHO cells tolerated this process condition.

[0084] As shown in Figure 6(a) and Figure 6(b), the bright area in the center of Figure 6(b) is a live cell. By counting the cells in the field of view and combining the field of view size parameters, the cell viability, cell density, live cell density and other parameters can be calculated. In Figure 6(a), the black outline and background near the periphery of each cell are formed by the part of the imaging beam corresponding to the transmission signal, while the central brightness of the live cell in Figure 6(b) and the granularity are formed by the part of the imaging beam corresponding to the scattered signal.

[0085] Furthermore, according to the results of real-time monitoring, the following Figure 7 By using the optical imaging sensor disclosed in the present disclosure, real-time analysis of cell size and granularity can be achieved without other processing. The image on the detector contains the cell morphology information (diameter) and cell surface information (granularity), and then the corresponding algorithm can be used to construct the (diameter-granularity) information. Figure 7 The scatter plot shown is used to achieve classification and clustering of cells.

[0086] In the optical imaging sensor and particle analysis equipment disclosed in the present invention, continuous sampling analysis can be realized in combination with a circulation pool. In addition to providing statistical information on the particles to be analyzed, detailed particle image information can also be supplemented, including information such as cell morphology and subcellular localization. By statistically analyzing the diameter and particle size information and combining it with the imaging information, the signal distribution within or between cells can be located and quantitatively analyzed. Thereby, statistical information on particle grouping and rich particle morphology and detailed information can be obtained in a single detection. In addition, the particle analysis disclosed in the present invention does not require complex preprocessing of the sample. The processor or computer can automatically extract the target signal from the obtained imaging information, eliminate interference such as debris, impurities, and reagent residues, and improve the effect of particle analysis.

[0087] 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.

[0088] 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.

[0089] 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.

[0090] 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.

[0091] 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.

[0092] 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.

[0093] 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.

[0094] 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 includes a diaphragm assembly disposed on the imaging optical path, and the diaphragm assembly includes: A first diaphragm having a first light-shielding portion and a first light-transmitting portion, and the first light-transmitting portion is configured to allow at least part of the light beam in the first region in the imaging light beam to pass through when participating in imaging, so as to form a first image; and A second diaphragm having a second light-shielding portion and a second light-transmitting portion, and the second light-transmitting portion is configured to allow at least part of the light beam in the second region different from the first region in the imaging light beam to pass through when participating in imaging, so as to form a second image, wherein the relative position of the second light-transmitting portion on the second diaphragm is different from the relative position of the first light-transmitting portion on the first diaphragm.

2. The optical imaging sensor according to claim 1, characterized in that: The first light-shielding portion is arranged to surround the first light-transmitting portion; and The second light-transmitting portion is arranged to surround the second light-shielding portion.

3. The optical imaging sensor according to claim 1, characterized in that: The relative position of the first light-transmitting portion on the first diaphragm is equivalent to the relative position of the second light-shielding portion on the second diaphragm; and The relative position of the first light-shielding portion on the first diaphragm is equivalent to the relative position of the second light-transmitting portion on the second diaphragm.

4. The optical imaging sensor according to claim 1, characterized in that: The optical imaging sensor further includes: A condenser lens disposed on the incident optical path, and the condenser lens is configured to converge the incident light beam; and An objective lens disposed on the imaging optical path, and the objective lens is configured to collect the imaging light beam.

5. The optical imaging sensor according to claim 4, characterized in that: NAc < NAo, where NAc is the numerical aperture of the condenser lens and NAo is the numerical aperture of the objective lens.

6. The optical imaging sensor according to claim 5, characterized in that: The first diaphragm is located at the rear focal plane of the objective lens when participating in imaging, the first light-transmitting portion is circular and located in the central region of the first diaphragm, and the radius r11 of the first light-transmitting portion satisfies r11 = NAc * f; and / or The second diaphragm is located at the rear focal plane of the objective lens when participating in imaging, the second light-shielding portion is circular and located in the central region of the second diaphragm, and the radius r22 of the second light-shielding portion satisfies r22 = NAc * f; where f is the focal length of the objective lens.

7. The optical imaging sensor according to claim 5, characterized in that: The first diaphragm is located between the rear focal plane and the exit pupil plane of the objective lens when participating in imaging, the first light-transmitting portion is circular and located in the central region of the first diaphragm, and the radius r11 of the first light-transmitting portion satisfies r11 = NAc * (f - s1); and / or The second diaphragm is located between the rear focal plane and the exit pupil plane of the objective lens when participating in imaging, the second light-shielding portion is circular and located in the central region of the second diaphragm, and the radius r22 of the second light-shielding portion satisfies r22 = NAc * (f - s2); where f is the focal length of the objective lens, s1 is the distance between the first diaphragm and the objective lens when participating in imaging, and s2 is the distance between the second diaphragm and the objective lens when participating in imaging.

8. The optical imaging sensor according to claim 1, characterized in that: The optical imaging sensor further includes: A switching member is mechanically connected to the first aperture and the second aperture, and is configured to switch one of the first aperture and the second aperture to be located on the imaging optical path, and to switch the other of the first aperture and the second aperture to be away from the imaging optical path.

9. The optical imaging sensor according to claim 8, characterized in that: The optical imaging sensor further comprises: An imaging mirror, wherein the imaging mirror is located on an exit light path of the objective lens, and the aperture assembly is located between the objective lens and the imaging mirror; and A detector is located on the output light path of the imaging mirror, and the detector is configured to perform imaging based on a portion of the imaging light beam.

10. The optical imaging sensor according to claim 1, characterized in that: The optical imaging sensor further comprises: a beam splitter, the beam splitter being located on an exit optical path of the objective lens and configured to allow a portion of the imaging light beam to travel along a first optical path and another portion of the imaging light beam to travel along a second optical path; The first optical path and the second optical path intersect each other, the first aperture is located on the first optical path, and the second aperture is located on the second optical path.

11. The optical imaging sensor according to claim 10, characterized in that: The optical imaging sensor further comprises: A first imaging mirror, wherein the first imaging mirror is located on an exit light path of the first aperture; a first detector, the first detector being located on an exit light path of the first imaging mirror, and the first detector being configured to perform imaging based on a portion of the imaging light beam; A second imaging mirror, the second imaging mirror is located on the exit light path of the second aperture; and A second detector is located on the output light path of the second imaging mirror, and the second detector is configured to perform imaging based on a portion of the imaging light beam.

12. The optical imaging sensor according to claim 1, characterized in that: The optical imaging sensor also includes a light source configured to generate an incident light beam.

13. A particle analysis device, characterized in that: The particle analysis equipment comprises: flow cell; and The optical imaging sensor according to any one of claims 1 to 12, wherein the optical imaging sensor is configured to generate a first imaging and a second imaging based on the sample in the flow cell.

14. The particle analysis device according to claim 13, characterized in that The particle analysis device also includes: Memory; and A processor is configured to analyze imaging data from the optical imaging sensor to generate target data.

Images