Cell image acquisition device and method

Abstract

The invention relates to the technical field of cells, and provides a cell image acquisition device and method. According to the device, a groove is formed in one side of a culture container, and a protruding part facing an opening of the groove is arranged at the bottom of the groove; the first reflecting device and the second reflecting device are arranged on a connecting surface of the side surface of the lug boss and the side surface of the groove, and a first reflecting surface of the first reflecting device and a second reflecting surface of the second reflecting device are oppositely arranged; light rays of the light source can be reflected by the first reflecting surface, penetrate through the convex part, reach the second reflecting surface, and reach the imaging lens after being reflected by the second reflecting surface, so as to collect a cell image of cell sap in the culture container. According to the cell image acquisition device provided by the invention, the cell image of the cell sap can be acquired by utilizing the reflection of light, so that the cell sampling is completed, an external device is not adopted to contact the cell sap in the whole process, the pollution to the cell sap is avoided, and the adverse effect on the cell growth is avoided.

Description

Technical Field

[0001] The present application relates to the field of cell technology, and in particular to a cell image acquisition device and method. Background Art

[0002] At present, in the process of cell culture using fully automatic cell culture equipment, it is necessary to detect the cell growth status during the culture process on time to achieve the purpose of process quality control. Among the detection targets, the most common one is cell density.

[0003] In order to detect the cell density, it is usually necessary to sample and count the cells to be tested, and the cell density is obtained based on the ratio of the number of cells in a specific volume of cell fluid and the specific volume.

[0004] However, the traditional method of cell sampling is to manually sample the cell fluid through a sterile needleless connector, which is equivalent to manually sampling the cells in the cell fluid. The sterile connector used in this technology will generate a negative pressure inside the connector when plugging and unplugging, thereby preventing foreign contaminants from entering the cell fluid. However, since some cell fluid will still remain at the connector, and this process cannot guarantee that external contaminants will not stick to the surface of the connector after plugging and unplugging, and enter the culture container with the residual cell fluid during the next use, contaminating the cell fluid, thereby affecting the growth of cells therein. Summary of the invention

[0005] The embodiments of the present application provide a cell image acquisition device and method to solve the technical problem that the traditional cell sampling method introduces foreign pollutants, causing cell fluid to be contaminated, thereby affecting cell growth.

[0006] In a first aspect, an embodiment of the present application provides a cell image acquisition device, comprising: a culture container, at least two reflective devices, a light source, and an imaging lens;

[0007] A groove is provided on one side of the culture container, a convex portion facing the opening of the groove is provided at the bottom of the groove, and a side surface of the convex portion is made of a light-transmitting material;

[0008] The reflecting device comprises a first reflecting device and a second reflecting device;

[0009] The first reflecting device and the second reflecting device are arranged on the connecting surface of the side surface of the protruding portion and the side surface of the groove, and the first reflecting surface of the first reflecting device and the second reflecting surface of the second reflecting device are arranged opposite to each other;

[0010] The first reflecting surface and the second reflecting surface respectively form acute angles with the plane where the opening of the groove is located. The light source and the imaging lens are arranged on one side of the opening of the groove, so that the light from the light source can be reflected by the first reflecting surface, pass through the protrusion, and reach the second reflecting surface, and then be reflected by the second reflecting surface to reach the imaging lens, so as to collect the cell image of the cell fluid in the culture container.

[0011] In one embodiment, the connecting surface is a surface inclined relative to the plane where the opening of the groove is located;

[0012] The first reflective device and the second reflective device are planar reflective devices, and the planar reflective devices are arranged on the connecting surface.

[0013] In one embodiment, the connection surface is a surface parallel to the plane where the opening of the groove is located;

[0014] The first reflecting device and the second reflecting device are right-angle reflecting mirrors, and the right-angle reflecting mirrors are arranged on the connecting surface, the first reflecting surface is the inclined surface of the first reflecting device, and the second reflecting surface is the inclined surface of the second reflecting device.

[0015] In one embodiment, it further comprises: a connecting bracket;

[0016] The connecting bracket is fixedly connected between the first reflecting device and the second reflecting device.

[0017] In one embodiment, it further comprises: a partition;

[0018] The partition is fixedly connected to the connecting bracket, and the partition is located on a connecting line between the light source and the second reflecting device.

[0019] In one embodiment, a surface of the protrusion away from the opening of the groove is a notched surface;

[0020] The width of the notch surface is less than or equal to 1 mm;

[0021] The width of the notch surface is the length of a line segment in the notch surface that is perpendicular to an incident light ray, and the incident light ray is light emitted by the light source and enters the protrusion after being reflected by the first reflector.

[0022] In one embodiment, the opening direction of the groove is perpendicular to the horizontal plane and downward, and the horizontal heights of the light source and the imaging lens are lower than the height of the plane where the opening of the groove is located;

[0023] The connecting surface and the notch surface are both surfaces inclined relative to the plane where the opening of the groove is located.

[0024] In a second aspect, an embodiment of the present application provides a cell image acquisition method, which is implemented using the cell image acquisition device described in the first aspect, and includes:

[0025] Controlling the light source to emit light to the first reflecting device to obtain a first reflected light;

[0026] controlling the imaging lens to receive the second reflected light to obtain a cell image;

[0027] The second reflected light is the light that the first reflected light reaches the second reflecting device after passing through the cell fluid in the protrusion and is then reflected by the second reflecting device.

[0028] In one embodiment, after obtaining the cell image, the method comprises:

[0029] Identifying and counting cells in the cell image using a pre-trained neural network model to obtain the number of cells in the cell image;

[0030] The cell density of the cell fluid is obtained according to the cell quantity and the parameter information of the cell image.

[0031] In one embodiment, obtaining the cell density of the cell fluid according to the cell number and the parameter information of the cell image includes:

[0032] Calculating the product of the area of ​​the cell image and the depth of field to obtain a sampling volume;

[0033] The ratio of the number of cells to the sample volume is calculated to obtain the cell density of the cell fluid.

[0034] The cell image acquisition device provided by the present application includes: a culture container, at least two reflective devices, a light source and an imaging lens, a groove is provided on one side of the culture container, a convex portion facing the opening of the groove is provided at the bottom of the groove, and the side of the convex portion is a light-transmitting material, the reflective device includes a first reflective device and a second reflective device, the first reflective device and the second reflective device are arranged on the connecting surface of the side of the convex portion and the side of the groove, and the first reflective surface of the first reflective device and the second reflective surface of the second reflective device are arranged oppositely, and the first reflective surface and the second reflective surface are respectively at an acute angle with the plane where the opening of the groove is located, and the light source and the imaging lens are arranged on one side of the opening of the groove, so that the light of the light source can be reflected by the first reflective surface, pass through the convex portion, and reach the second reflective surface, and then reflect by the second reflective surface to reach the imaging lens, so as to collect the cell image of the cell fluid in the culture container. The cell image acquisition device of the present application can collect the cell image of the cell fluid by the reflection of light, so as to complete the sampling of the cell, and no external device is used to contact the cell fluid in the whole process, thereby preventing the contamination of the cell fluid, thereby preventing the adverse effect on the growth of the cell. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the technical solutions in the present application or the prior art, a brief introduction will be given below to the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0036] Figure 1 It is one of the front views of the cell image acquisition device provided in the embodiment of the present application;

[0037] Figure 2 This is the second front view of the cell image acquisition device provided in the embodiment of the present application;

[0038] Figure 3 is a three-dimensional diagram of the connection structure of the connection bracket and the right-angle reflector provided in an embodiment of the present application;

[0039] Figure 4 is a stereoscopic diagram of a culture container provided in an embodiment of the present application;

[0040] Figure 5 yes Figure 1 Another configuration diagram of the structure of the method;

[0041] Figure 6 This is one of the flow charts of the cell image acquisition method provided in the embodiment of the present application;

[0042] Figure 7 This is the second flow chart of the cell image acquisition method provided in the embodiment of the present application.

[0043] Reference numerals:

[0044] 1-culture container; 2-plane reflective device; 3-right-angle reflector; 4-light source; 5-imaging lens; 6-groove; 7-raised portion; 8-connecting surface; 9-connecting bracket; 10-notch surface; 11-partition. DETAILED DESCRIPTION

[0045] In order to make the purpose, technical solutions and advantages of this application clearer, the technical solutions in this application will be clearly and completely described below in conjunction with the drawings in the embodiments of this application. Obviously, the described embodiments are part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0046] Figure 1 It is one of the front views of the cell image acquisition device provided in the embodiment of the present application;

[0047] Figure 2 This is the second front view of the cell image acquisition device provided in the embodiment of the present application;

[0048] Reference Figure 1 to Figure 2 , the embodiment of the present application provides a cell image acquisition device, which may include: a culture container 1, at least two reflective devices, a light source 4 and an imaging lens 5;

[0049] A groove 6 is provided on one side of the culture container 1, a convex portion 7 facing the opening of the groove 6 is provided at the bottom of the groove 6, and the side surface of the convex portion 7 is made of light-transmitting material;

[0050] The reflecting device comprises a first reflecting device and a second reflecting device;

[0051] The first reflector and the second reflector are arranged on the connecting surface 8 of the side surface of the protrusion 7 and the side surface of the groove 6, and the first reflective surface of the first reflector and the second reflective surface of the second reflector are arranged opposite to each other;

[0052] The first reflecting surface and the second reflecting surface respectively form acute angles with the plane where the opening of the groove 6 is located. The light source 4 and the imaging lens 5 are arranged on the opening side of the groove 6, so that the light from the light source 4 can be reflected by the first reflecting surface and then pass through the protrusion 7 and reach the second reflecting surface, and then reflected by the second reflecting surface and reach the imaging lens 5, so as to collect the cell image of the cell fluid in the culture container 1.

[0053] The culture container 1 is sealed except for the input and output ports of the cell fluid;

[0054] The culture container 1 can be placed with the opening of the groove 6 facing upward, downward, to the left, to the right, or in other directions, without limitation here. When the placement direction of the culture container 1 is different, the light source 4 and the imaging lens 5 also change accordingly with the change of the opening direction of the groove 6.

[0055] In this embodiment, the side surface of the protrusion 7 is made of a light-transmitting material which may be polyvinyl chloride material or other materials, which are not limited here, as long as the light can be completely transmitted.

[0056] In addition, the culture container 1 is placed with the opening of the groove 6 facing upward, and the light source 4 and the imaging lens 5 are both located directly above the two reflective devices, so as to control the light source 4 to emit light downward, and the light is reflected twice by the two reflective devices and then enters the imaging lens 5 upward.

[0057] It should be noted that if the height of the protrusion 7 is too low or the horizontal height of the reflection points on the two reflective devices is high, the light emitted by the light source 4 may pass through the top of the protrusion 7 after being reflected by the first reflective device. Since the light does not pass through the cell fluid in the protrusion 7, even if the imaging lens 5 receives the light reflected by the second reflective device, it cannot image the cell fluid in the culture container 1.

[0058] Therefore, the height of the protrusion 7, the specific position of the reflector, and the specific positions of the light source 4 and the imaging lens 5 need to be adjusted in coordination to ensure that the light emitted by the light source 4 can pass through the protrusion 7 to reach the second reflector after being reflected by the first reflector, and then be reflected by the second reflector to the imaging lens 5.

[0059] In order to facilitate imaging control and achieve better imaging effects, the angle between the first reflecting surface, the second reflecting surface and the plane where the opening of the groove 6 is located can be set to 45 degrees. In this way, it is only necessary to control the light source 4 to emit light perpendicular to the plane where the opening of the groove 6 is located to the first reflecting device. The light reflected by the first reflecting device will reach the second reflecting device parallel to the plane where the opening of the groove 6 is located. The second reflecting device will then reflect the light perpendicular to the plane where the opening of the groove 6 is located, and then control the imaging lens 5 to receive the reflected light perpendicular to the plane where the opening of the groove 6 is located.

[0060] The cell image acquisition device provided in this embodiment includes: a culture container, at least two reflective devices, a light source and an imaging lens, a groove is provided on one side of the culture container, a protrusion facing the opening of the groove is provided at the bottom of the groove, and the side of the protrusion is a light-transmitting material, the reflective device includes a first reflective device and a second reflective device, the first reflective device and the second reflective device are arranged on the connecting surface of the side of the protrusion and the side of the groove, and the first reflective surface of the first reflective device and the second reflective surface of the second reflective device are arranged oppositely, and the first reflective surface and the second reflective surface are respectively at an acute angle with the plane where the opening of the groove is located, and the light source and the imaging lens are arranged on one side of the opening of the groove, so that the light of the light source can be reflected by the first reflective surface, pass through the protrusion, and reach the second reflective surface, and then reach the imaging lens after being reflected by the second reflective surface, so as to collect the cell image of the cell fluid in the culture container. The cell image acquisition device in this embodiment can collect the cell image of the cell fluid by the reflection of light, so as to complete the sampling of the cell, and no external device is used to contact the cell fluid in the whole process, thereby preventing the contamination of the cell fluid, thereby preventing the adverse effect on the cell growth.

[0061] Furthermore, since the present embodiment directly performs cell imaging on the cell fluid in the culture container, there is no error in physical sampling and counting, thereby improving the reliability of cell counting;

[0062] Furthermore, since the present embodiment does not require actual extraction of the cell fluid in the culture container, sample loss in the traditional sampling process is avoided.

[0063] Reference Figure 1 In one embodiment, the connecting surface 8 is a surface inclined relative to the plane where the opening of the groove 6 is located;

[0064] The first reflective device and the second reflective device are planar reflective devices 2 , and the planar reflective devices 2 are arranged on the connecting surface 8 .

[0065] The first reflective device and the second reflective device can be paper or film with a reflective function, such as tin foil, or a plane mirror, which is not limited here.

[0066] It should be noted that the first reflective device or the second reflective device does not need to completely cover the entire connecting surface 8 , but only needs to cover the incident point of the light source 4 , which helps to save reflective device materials without affecting the reflective function.

[0067] Furthermore, the angle between the connecting surface 8 and the plane where the opening of the groove 6 is located can be 45 degrees, so that the angle between the first reflecting surface of the first reflecting device and the second reflecting surface of the second reflecting device on the connecting surface 8 and the plane where the opening of the groove 6 is located is 45 degrees, which facilitates imaging control to achieve better imaging effects.

[0068] This embodiment sets the connection surface as a surface inclined relative to the plane where the opening of the groove is located, and cooperates with a planar reflective device to reflect the incident light twice, and finally completes the cell imaging of the cell fluid of the protrusion by the imaging lens.

[0069] Reference Figure 2 In one embodiment, the connecting surface 8 is a surface parallel to the plane where the opening of the groove 6 is located;

[0070] The first reflecting device and the second reflecting device are right-angle reflecting mirrors 3, and the right-angle reflecting mirrors 3 are arranged on the connecting surface 8, the first reflecting surface is the inclined surface of the first reflecting device, and the second reflecting surface is the inclined surface of the second reflecting device.

[0071] Furthermore, the first reflecting device and the second reflecting device may be isosceles right-angle reflecting mirrors.

[0072] Since the inclined surface of the isosceles right-angle reflector is a reflective surface and the angle between it and each right-angle surface must be 45 degrees, the angle between the first reflective surface of the first reflective device on the connecting surface 8 and the second reflective surface of the second reflective device and the plane where the opening of the groove 6 is located is 45 degrees, which facilitates imaging control to achieve better imaging effects.

[0073] It should be noted that the size of the right-angle reflector 3 or the size of the reflective surface can also be adjusted to ensure that it can cover the incident point of the light source 4, which helps to save reflective device materials without affecting the reflective function.

[0074] In addition, when the culture container 1 is placed with the opening of the groove 6 facing upward, setting the connecting surface 8 to be parallel to the plane where the opening of the groove 6 is located is easier to process than setting it to be inclined relative to the plane where the opening of the groove 6 is located. Moreover, when a reflective device is set on a connecting surface inclined relative to the plane where the opening of the groove 6 is located, if the reflective device is small in size, the reflective device is more likely to slide on the connecting surface due to unbalanced force, and additional fixing means are required to fix the reflective device on the connecting surface. However, when a reflective device is set on a connecting surface parallel to the plane where the opening of the groove 6 is located, no matter whether the size of the reflective device is large or small, the force balance can be better maintained without the need for additional fixing means to fix the reflective device on the connecting surface.

[0075] This embodiment sets the connecting surface to be parallel to the plane where the opening of the groove is located, and cooperates with a right-angle reflector to reflect the incident light twice, and finally the imaging lens completes the cell imaging of the cell fluid in the raised part. At the same time, it can make the culture container easier to process and the reflective device easier to fix.

[0076] Figure 3 is a three-dimensional diagram of the connection structure of the connection bracket and the right-angle reflector provided in the embodiment of the present application; Figures 2 to 3 , in one embodiment, the cell image acquisition device may further include a connecting bracket 9;

[0077] The connecting bracket 9 is fixedly connected between the first reflecting device and the second reflecting device.

[0078] Furthermore, in order to improve the structural strength of the connecting bracket 9, the connecting bracket 9 can be set as a rectangular frame, in which four parallel frame strips extend toward the right-angle reflector 3 and are connected to two right-angle surfaces opposite to the right-angle reflector 3.

[0079] When the groove 6 of the culture container 1 is small or the depth of the groove 6 is deep, it is inconvenient to take out and install the right-angle reflector 3 placed therein. Therefore, a connecting bracket 9 can be used to connect the two right-angle reflectors 3, and the two right-angle reflectors 3 can be taken out of the groove 6 or installed into the groove 6 by controlling the movement of the connecting bracket 9.

[0080] It should be noted that when the reflector assembly formed by connecting the connecting bracket 9 and the two right-angle reflectors 3 is installed in the groove 6, in order to fix it firmly, the size of the reflector assembly is usually designed to be compatible with the shape inside the groove 6 so that it can be embedded in the groove 6 to achieve a better fixing effect. When installing the reflector assembly, a large force will be continuously applied to the connecting bracket 9 to push the entire reflector assembly into the groove 6 until it is finally completely embedded. In this process, the rectangular connecting bracket 9 can withstand a large applied force due to its stable structure, and will not be deformed during the force application process, thereby affecting the embedding of the right-angle reflector 3.

[0081] In addition, no matter what form the connecting bracket 9 is configured in, it must not affect the path of light propagation.

[0082] In this embodiment, a connecting bracket is provided to connect two right-angle reflectors to form a reflector assembly, and the right-angle reflector can be conveniently and quickly installed in the groove of the culture container or removed from the groove of the culture container by controlling the movement of the connecting bracket.

[0083] Reference Figures 2 to 3 , In one embodiment, the cell image acquisition device further includes: a partition 11;

[0084] The partition 11 is fixedly connected to the connecting bracket 9 , and the partition 11 is located on the connecting line between the light source 4 and the second reflecting device.

[0085] Although when the light source 4 is controlled to emit light, the light is usually controlled to focus on the first reflective device, there will still be a small amount of light divergence, which may reach the second reflective device and affect the imaging. Therefore, a partition 11 can be set to block the path of light to the second reflective device to reduce interference with imaging. The setting position of the partition 11 can be adjusted according to the relative position of the light source 4 and the second reflective device, and is not limited here.

[0086] In this embodiment, by arranging a partition on the connecting bracket, the path of light leading to the second reflective device can be blocked to reduce interference with imaging.

[0087] Figure 4 is a stereogram of a culture container provided in an embodiment of the present application; Figure 1 to Figure 2 ,as well as Figure 4 In one embodiment, a surface of the protrusion 7 away from the opening of the groove 6 is a notch surface 10;

[0088] The width of the notch surface 10 is less than or equal to 1 mm;

[0089] The width of the notch surface 10 is the length of a line segment D in the notch surface 10 that is perpendicular to the incident light. The incident light is the light emitted by the light source 4 and enters the protrusion 7 after being reflected by the first reflector.

[0090] In this embodiment, the width of the notch surface is limited to no more than 1 mm to prevent the width from being too large, resulting in excessive cell fluid in the protrusion, so that the incident light is absorbed by the cell fluid in large quantities, and the outgoing light becomes blurred, affecting the cell imaging effect of the cell fluid.

[0091] Figure 5 yes Figure 1 Another configuration diagram of the structure of the method;

[0092] Reference Figure 5 In one embodiment, the opening direction of the groove 6 is perpendicular to the horizontal plane downward, and the horizontal heights of the light source 4 and the imaging lens 5 are lower than the height of the plane where the opening of the groove 6 is located;

[0093] Both the connecting surface 8 and the notch surface 10 are surfaces inclined relative to the plane where the opening of the groove 6 is located.

[0094] Furthermore, the inclination angle of the notch surface 10 relative to the plane where the opening of the groove 6 is located is greater than or equal to 5 degrees.

[0095] Figure 5 and Figure 1 In the figure, the light source 4 and the imaging lens 5 are swapped with the horizontal height of the culture container 1.

[0096] If adopted Figure 1 In the arrangement mode, cells will settle to the bottom of the culture container 1 due to gravity. Therefore, the cell fluid in the cell fluid near the top of the culture container 1 will have fewer cells, and the cell fluid near the bottom will have more cells. Therefore, the depth of the groove 6 needs to be designed to be deeper, so that the cell fluid in the raised portion 7 is the cell fluid at the medium depth of the culture container 1, thereby ensuring that the number of cells in the imaging picture is moderate and improving the reliability of cell imaging. At the same time, the culture container 1 can also be placed in a rocking reactor for rocking and mixing, and the flow field formed by the rocking can be used to flush the notch surface to avoid cell deposition. However, the deeper the groove 6 is, the more structural strength of the culture container 1 will be. Therefore, it can be used Figure 5 The setup method shown.

[0097] When using Figure 5 In the arrangement shown, the groove 6 does not need to be designed to be very deep, otherwise the cells will be deposited more quickly on the protrusion due to gravity. Although the depth of the groove 6 is reduced, the effect of reducing the deposition of cells on the protrusion 7 is still limited, so the notch surface 10 can be set as a surface inclined relative to the plane where the opening of the groove 6 is located, and the inclination angle of the notch surface 10 is adjusted. Since the connecting surface 8 is also a surface inclined relative to the plane where the opening of the groove 6 is located, the cells can slide from the connecting surface 8 that is first contacted to the side of the protrusion 7, thereby further reducing the deposition of cells on the protrusion 7.

[0098] This embodiment provides another setting method of the cell image acquisition device, which can improve the structural strength of the culture container. At the same time, by setting the notch surface to a surface inclined relative to the plane where the opening of the groove is located, the deposition of cells on the protrusion is reduced, thereby improving the reliability of cell imaging.

[0099] Figure 6 is one of the flow charts of the cell image acquisition method provided in the embodiment of the present application; Figure 6 The present application embodiment provides a cell image acquisition method, which is implemented using the aforementioned cell image acquisition device and may include:

[0100] 601. Control the light source to emit light to the first reflecting device to obtain a first reflected light;

[0101] 602. Control the imaging lens to receive the second reflected light to obtain a cell image;

[0102] The second reflected light is the light that the first reflected light reaches the second reflecting device after passing through the cell fluid in the protrusion and is then reflected by the second reflecting device.

[0103] The cell image can be the highest definition image or the image with the clearest cell boundary selected after clarity recognition from among the multiple images collected during the automatic focusing process of the imaging lens. The imaging lens can magnify the image during the automatic focusing process, and the magnification can be adjusted according to actual needs, which is not limited here. In this embodiment, the magnification can be set to 40 times.

[0104] The cell image acquisition method provided in this embodiment starts the light source and controls the light source to emit light to the first reflector. The light passes through the cell fluid in the protrusion after being reflected by the first reflector and reaches the second reflector. The light is then received by the imaging lens after being reflected by the second reflector. At this point, the imaging lens completes the cell image acquisition of the cell fluid in the protrusion. The cell image acquisition method in this embodiment can collect the cell image of the cell fluid by the reflection of light, thereby completing the sampling of cells. No external device is used to contact the cell fluid during the whole process, thereby eliminating the contamination of the cell fluid and thus eliminating the adverse effects on cell growth.

[0105] Furthermore, since the present embodiment directly performs cell imaging on the cell fluid in the culture container, there is no error in physical sampling and counting, thereby improving the reliability of cell counting;

[0106] Furthermore, since the present embodiment does not require actual extraction of the cell fluid in the culture container, sample loss in the traditional sampling process is avoided.

[0107] Figure 7This is the second flow chart of the cell image acquisition method provided in the embodiment of the present application; Figure 7 In one embodiment, after obtaining the cell image, the method may include:

[0108] 701. Identify and count cells in the cell image using a pre-trained neural network model to obtain the number of cells in the cell image;

[0109] 702. Calculate the product of the area of ​​the cell image and the depth of field to obtain the sampling volume;

[0110] 703. Calculate the ratio of the number of cells to the sample volume to obtain the cell density of the cell fluid.

[0111] In step 701, the cell image can be first input into a pre-trained neural network model, and the cell shape features of the cell image can be extracted through the model, and the cells can be classified according to the extracted cell shape features, and the cells with the cell shape features can be classified. If the probability of the cell being classified as the cell shape feature is greater than a preset threshold, the cell is marked as a cell to be counted, and the number of cells to be counted is counted to obtain the number of cells in the cell image. The network structure of the neural network model can be a network structure such as YOLO, VGG or ResNets, and the cell shape features include the size, shape, agglomeration, and stacking of the cell.

[0112] The YOLO (You Only Look Once, YOLO) network structure is an algorithmic network structure that provides real-time object detection. The YOLO algorithm uses a convolutional neural network to perform a forward propagation to detect objects, and can detect and identify various objects in the picture in real time. Object detection in YOLO is completed as a regression problem and provides the category probability of the detected object.

[0113] The Visual Geometry Group (VGG) network structure uses three 3×3 convolution kernels instead of 7×7 convolution kernels, and two 3×3 convolution kernels instead of 5×5 convolution kernels. The main purpose of this is to increase the depth of the network while ensuring the same receptive field, thereby improving the effect of the neural network to a certain extent.

[0114] The Residual Networks (ResNets) structure allows some layers of the neural network to skip the connection with the next layer of neurons, connecting the layers and weakening the strong connection between each layer. This helps solve the gradient disappearance and gradient explosion problems, and can ensure good network performance while training deeper networks.

[0115] In step 702, the area and depth of field of the cell image can be obtained through the setting information of the imaging lens, and the setting information is parameter information pre-configured for the imaging lens.

[0116] It should be noted that the depth of field of the cell image is smaller than the width of the notch surface.

[0117] This embodiment uses a pre-trained neural network model to identify and count cells in a cell image, and can reliably measure parameters such as cell size, roundness, and agglomeration rate. At the same time, the sampling volume is obtained through the area and depth of field of the cell image, and the cell density of the cell fluid is obtained through the ratio of the number of cells to the sampling volume. The cell density can be measured more accurately, and since there is no need to stain the cells during the entire process, the contamination of the cells by traditional cell counting methods is avoided.

[0118] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit it. Although the present application has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A cell image acquisition device, characterized in that: include: A culture container, at least two reflective devices, a light source, and an imaging lens; A groove is provided on one side of the culture container, a convex portion facing the opening of the groove is provided at the bottom of the groove, and a side surface of the convex portion is made of a light-transmitting material; The reflecting device comprises a first reflecting device and a second reflecting device; The first reflecting device and the second reflecting device are arranged on the connecting surface of the side surface of the protruding portion and the side surface of the groove, and the first reflecting surface of the first reflecting device and the second reflecting surface of the second reflecting device are arranged opposite to each other; The first reflecting surface and the second reflecting surface respectively form acute angles with the plane where the opening of the groove is located. The light source and the imaging lens are arranged on one side of the opening of the groove, so that the light from the light source can be reflected by the first reflecting surface, pass through the protrusion, and reach the second reflecting surface, and then be reflected by the second reflecting surface to reach the imaging lens, so as to collect the cell image of the cell fluid in the culture container.

2. The cell image acquisition device according to claim 1, characterized in that: The connecting surface is a surface inclined relative to the plane where the opening of the groove is located; The first reflective device and the second reflective device are planar reflective devices, and the planar reflective devices are arranged on the connecting surface.

3. The cell image acquisition device according to claim 1, characterized in that: The connecting surface is a surface parallel to the plane where the opening of the groove is located; The first reflecting device and the second reflecting device are right-angle reflecting mirrors, and the right-angle reflecting mirrors are arranged on the connecting surface, the first reflecting surface is the inclined surface of the first reflecting device, and the second reflecting surface is the inclined surface of the second reflecting device.

4. The cell image acquisition device according to claim 3, characterized in that: Also includes: Connect the bracket; The connecting bracket is fixedly connected between the first reflecting device and the second reflecting device.

5. The cell image acquisition device according to claim 4, characterized in that: Also includes: Partitions; The partition is fixedly connected to the connecting bracket, and the partition is located on a connecting line between the light source and the second reflecting device.

6. The cell image acquisition device according to claim 1, characterized in that: A surface of the protrusion away from the opening of the groove is a notch surface; The width of the notch surface is less than or equal to 1 mm; The width of the notch surface is the length of a line segment in the notch surface that is perpendicular to an incident light ray, and the incident light ray is light emitted by the light source and enters the protrusion after being reflected by the first reflector.

7. The cell image acquisition device according to claim 6, characterized in that: The opening direction of the groove is perpendicular to the horizontal plane and downward, and the horizontal heights of the light source and the imaging lens are lower than the height of the plane where the opening of the groove is located; The connecting surface and the notch surface are both surfaces inclined relative to the plane where the opening of the groove is located.

8. A cell image acquisition method, characterized in that: The method is implemented by using the cell image acquisition device according to any one of claims 1 to 7, comprising: Controlling the light source to emit light to the first reflecting device to obtain a first reflected light; controlling the imaging lens to receive the second reflected light to obtain a cell image; The second reflected light is the light that the first reflected light reaches the second reflecting device after passing through the cell fluid in the protrusion and is then reflected by the second reflecting device.

9. The cell image acquisition method according to claim 8, characterized in that: After obtaining the cell image, the method comprises: Identifying and counting cells in the cell image using a pre-trained neural network model to obtain the number of cells in the cell image; The cell density of the cell fluid is obtained according to the cell quantity and the parameter information of the cell image.

10. The cell image acquisition method according to claim 9, characterized in that: The step of obtaining the cell density of the cell fluid according to the cell quantity and the parameter information of the cell image includes: Calculating the product of the area of ​​the cell image and the depth of field to obtain a sampling volume; The ratio of the number of cells to the sample volume is calculated to obtain the cell density of the cell fluid.

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