Confirmation method, biological detection system, and storage medium

Abstract

The invention discloses a confirmation method, a biological detection system and a storage medium. The invention provides a confirmation method, which is used for confirming the affiliation of an optical signal of a test sample, and comprises the following steps: controlling a light source to project non-uniform light to a surface of the test sample so as to excite the test sample to form the optical signal on the surface; acquiring a speckle image formed based on the optical signal through an optical sensor; based on the speckle image, an affiliation of the optical signal is determined, the affiliation of the optical signal including whether the optical signal includes a non-target optical signal. Therefore, non-uniform light is projected to the surface of the test sample to form the speckle image, and the attribution of the optical signal of the test sample is confirmed through the speckle image, so that whether the surface of the test sample has defects can be judged, the quality of the test sample is ensured, and the data quality of optical detection based on the test sample is further ensured.

Description

Technical Field

[0001] The present invention relates to the technical field of optical detection, and in particular, to a confirmation method, a biological detection system, and a storage medium. Background Art

[0002] Gene sequencing technology refers to the technical means of obtaining the DNA or RNA base sequence through detection. Currently, the dominant sequencing technology is high-throughput sequencing technology. In high-throughput sequencing technology, the general process of gene sequencing includes fixing a nucleic acid sample to be tested on a biochip in a way such as hybridization, then using PCR amplification to form nucleic acid molecule clusters of the nucleic acid sample to be tested, and then adding bases with fluorescent groups, polymerase, primers, etc. Through the principle of base complementary pairing, the bases with fluorescent groups are combined with the bases on the nucleic acid sample to be tested. Finally, the fluorescent groups are excited by an optical imaging system to generate fluorescence and the fluorescence is collected to form an image, and base recognition is performed through the image, so as to realize the determination of the base sequence of the nucleic acid sample to be tested.

[0003] When preparing a biochip, it is necessary to judge whether there are defects on the upper and lower surfaces of the flow channel in the biochip, such as uneven brightness, modified bubbles, bright lines, etc., to ensure the quality of the chip, and further ensure the data quality of optical detection based on the test sample. Summary of the Invention

[0004] The present invention provides a confirmation method, a biological detection system, and a storage medium.

[0005] An embodiment of the present application provides a confirmation method, which is used to confirm the attribution of the optical signal of a test sample, and includes: controlling a light source to project uneven light onto one surface of the test sample to excite the test sample to form an optical signal on the surface; obtaining a speckle image formed based on the optical signal through an optical sensor; determining the attribution of the optical signal based on the speckle image, and the attribution of the optical signal includes whether the optical signal includes non-target optical signals.

[0006] In this way, by projecting uneven light onto the surface of the test sample to form a speckle image, and confirming the attribution of the optical signal of the test sample through the speckle image, it is possible to judge whether there are defects on the surface of the test sample, ensure the quality of the test sample, and further ensure the data quality of optical detection based on the test sample.

[0007] In some embodiments, the uneven light is light with uneven intensity and / or uneven distribution.

[0008] In this way, the uneven light irradiating the test sample can form a speckle image, which is convenient for confirming the attribution of the optical signal of the test sample.

[0009] In some embodiments, the light source includes a laser light source. Controlling the light source to project non-uniform light onto a surface of a test sample includes: controlling the laser light source to emit self-coherent light so that the self-coherent light interferes to form non-uniform light; and projecting the non-uniform light onto a surface of the test sample.

[0010] In this way, by controlling the laser light source to emit self-coherent light, non-uniform light can be formed, enabling a speckle image to be formed on the surface of the test sample, which is convenient for confirming the attribution of the optical signal of the test sample.

[0011] In some embodiments, determining the attribution of the optical signal based on the speckle image includes: when there is a bright spot in the speckle image, determining the intensity of the bright spot; comparing the intensity of the bright spot with a first preset intensity, and determining the bright spot with an intensity higher than the first preset intensity as a non-target optical signal; and / or when there is a dark spot in the speckle image, comparing the intensity of the dark spot with a second preset intensity, and determining the dark spot with an intensity lower than the second preset intensity as a non-target optical signal.

[0012] When the non-uniform light irradiates a surface of the test sample, a fluorescence signal of the test sample is excited. The excited fluorescence forms bright spots, and the unexcited fluorescence forms dark spots. When the optical signal generated on the surface of the test sample is a target optical signal, the intensity of the bright spots in the speckle image is lower than or equal to the first preset intensity, and the intensity of the dark spots is higher than or equal to the second preset intensity. When there is abnormal adsorption on the surface of the test sample, the brightness of the excited fluorescence changes, and the intensities of the dark spots and bright spots also change. If the intensity of the bright spots is higher than the first preset intensity or the intensity of the dark spots is lower than the second preset intensity, it can be indicated that the optical signal includes non-target optical signals. In this way, by comparing the intensity of the bright spots with the first preset intensity or the intensity of the dark spots with the second preset intensity, the attribution of the optical signal can be determined.

[0013] In some embodiments, the test sample includes a biochip, and no test sample to be detected is bound to the surface of the biochip.

[0014] In this way, by detecting the biochip with no test sample to be detected bound to its surface, it can be determined whether there are defects on the surface of the biochip and whether the biochip is qualified.

[0015] In some embodiments, the biochip includes a first substrate layer and a second substrate layer arranged in a stacked manner, and a connection layer arranged between the first substrate layer and the second substrate layer.

[0016] In this way, the first substrate layer and the second substrate layer of the biochip can be detected to ensure the quality of the biochip, and further ensure the data quality of the optical detection based on the test sample.

[0017] In some embodiments, the first substrate layer is a substrate layer with a light transmittance greater than or equal to 90%.

[0018] In this way, the first substrate layer has good light transmittance, enabling non-uniform light to enter the chip, and the optical signals inside the chip can also be emitted to the outside, resulting in a better imaging effect of the speckle image.

[0019] In some embodiments, controlling the light source to project non-uniform light onto a surface of the test sample includes: controlling the light source to project non-uniform light onto the surface where the first substrate layer of the biochip is located.

[0020] In this way, the non-uniform light can be projected onto the first substrate layer in a critical illumination manner to form a speckle image, facilitating the detection of the surface of the first substrate layer.

[0021] In some embodiments, the first substrate layer includes a B surface facing the second substrate layer. Obtaining a speckle image based on the optical signal through an optical sensor includes: obtaining a first speckle image based on the optical signal generated by the B surface through the optical sensor.

[0022] In this way, the B surface of the first substrate layer can be detected through the first speckle image.

[0023] In some embodiments, based on the speckle image, determining the attribution of the optical signal, where the attribution of the optical signal includes whether the optical signal includes non-target optical signals, includes: when there are dark spots in the first speckle image, comparing the intensity of the dark spots with a second preset intensity, and determining the dark spots with an intensity lower than the second preset intensity as non-target optical signals.

[0024] When there is no abnormal adsorption on the B surface, the fluorescence signal is excited by the non-uniform light to form speckles, and the first speckle image is arranged in a speckle pattern. When there is abnormal adsorption on the B surface, the fluorescence signal in the abnormal adsorption area is not excited and forms dark spots with lower intensity. If the intensity of the dark spots is lower than the second preset intensity, it can indicate that the fluorescence brightness excited by the B surface decreases and the number of fluorescence excited by the B surface decreases, that is, there is abnormal adsorption on the B surface. In this way, by comparing the intensity of the dark spots with the second preset intensity, the attribution of the optical signal can be determined, thereby detecting the B surface of the first substrate layer of the biochip.

[0025] In some embodiments, the second substrate layer includes a C surface facing the first substrate layer. Obtaining a speckle image based on the optical signal through an optical sensor includes: obtaining a second speckle image based on the optical signal generated by the C surface through the optical sensor.

[0026] In this way, the C surface of the second substrate layer can be detected through the second speckle image.

[0027] In some embodiments, based on the speckle image, the attribution of the optical signal is determined. The attribution of the optical signal includes whether the optical signal includes non-target optical signals, and it includes: dividing the second speckle image into multiple regions; if the average intensity of the region pixels is higher than the first preset intensity, it is determined that there are non-target optical signals in the optical signal of the C surface.

[0028] When there is no abnormal adsorption on the C surface, the second speckle image is an image of the background light reflection; when there is abnormal adsorption on the C surface, the adsorption amount of each region is different, so the excited fluorescence brightness is different, and the abnormal adsorption region forms a speckle, and the intensity of the speckle is higher than the first preset intensity. Thus, by comparing the average intensity of the region pixels of the second speckle image with the first preset intensity, the attribution of the optical signal can be determined, thereby detecting the C surface of the second substrate layer of the biochip.

[0029] In some embodiments, the optical signal is a fluorescence signal.

[0030] Thus, when the uneven light emitted by the light source irradiates the surface of the test sample, the fluorescence signal will emit light, and the emitted light converges to the optical sensor to form a speckle image.

[0031] In some embodiments, the method includes: arranging a light homogenizing device on the optical path of the light source to homogenize the light emitted by the light source, so as to project uniform light onto one surface of the test sample to excite the test sample to form an optical signal on the surface; acquiring a surface image based on the optical signal through an optical sensor; based on the surface image, determining the attribution of the optical signal, and the attribution of the optical signal includes whether the optical signal includes non-target optical signals.

[0032] Thus, by projecting uniform light onto the surface of the test sample to form a surface image, and by confirming the attribution of the optical signal of the test sample through the surface image, it can be judged whether there are defects on the surface of the test sample, ensuring the quality of the test sample, and further ensuring the data quality of the optical detection based on the test sample.

[0033] In some embodiments, determining the attribution of the optical signal based on the speckle image includes: when there are dark spots in the surface image, comparing the intensity of the dark spots with the second preset intensity, and determining the dark spots with intensity lower than the second preset intensity as non-target optical signals.

[0034] When there is no abnormal adsorption on the B surface of the test sample, the intensity of the surface image is the excited fluorescence brightness; when there is abnormal adsorption on the B surface of the test sample, the fluorescence signal in the abnormal adsorption region is not excited and forms a dark spot. If the intensity of the dark spot is lower than the second preset intensity, it can be explained that the optical signal generated on the B surface of the test sample includes non-target optical signals. Thus, by comparing the intensity of the dark spot with the second preset intensity, the attribution of the optical signal can be determined, thereby detecting the surface of the test sample.

[0035] In some embodiments, an objective lens and a collimator are disposed between the test sample and the light source. The collimator is configured to collimate the light emitted by the light source into parallel light; the objective lens is configured to converge the parallel light onto the surface of the test sample to form critical illumination on the surface of the test sample.

[0036] In this way, the collimator can collimate the non-uniform light from the light source into parallel light, so that the non-uniform light remains parallel during transmission, reducing energy loss and beam divergence. Critical illumination converges the parallel light onto the surface of the test sample, which can reduce the interference of the untested surface on the test surface and improve the imaging effect.

[0037] In some embodiments, a first dichroic mirror is disposed between the collimator and the objective lens. The first dichroic mirror is disposed on the optical axis of the objective lens. The first dichroic mirror is configured to reflect the parallel light to the objective lens and to transmit the optical signal from the surface of the test sample to the optical sensor.

[0038] In this way, the first dichroic mirror reflects the parallel light to the objective lens, so that the parallel light irradiates the test sample after passing through the objective lens to excite the surface of the test sample to form an optical signal.

[0039] In some embodiments, a second dichroic mirror is disposed on a side of the first dichroic mirror away from the objective lens. A focusing device is disposed on a side of the second dichroic mirror deviating from the optical axis of the objective lens. The focusing device is configured to emit a focusing beam. The focusing beam is reflected by the second dichroic mirror and propagates along the optical axis of the objective lens and irradiates the test sample. The focusing device is further configured to receive the focusing beam reflected from the test sample and reflected by the second dichroic mirror; the second dichroic mirror is further configured to transmit the optical signal from the first dichroic mirror to the optical sensor.

[0040] In this way, the focusing device is mainly configured to mark the object distance after the objective lens is focused, and to monitor the change amount of the object distance in real time during the optical detection based on the test sample, and correct it to ensure that the system is always in clear focus, ensure the accuracy of the image collected by the optical sensor, and finally obtain a clear image within the depth of focus.

[0041] In some embodiments, the light source is a multimode laser.

[0042] In this way, the laser emitted by the multimode laser irradiating the test sample can generate speckles, which is convenient for forming a speckle image.

[0043] An embodiment of the present application provides a biological detection system, including a memory and a processor. The processor is configured to execute a computer program stored in the memory to implement the method according to any one of the above embodiments.

[0044] Embodiments of the present application provide a non-volatile computer-readable storage medium storing a computer program, which implements the method according to any of the above embodiments when the computer program is executed by one or more processors.

[0045] Additional aspects and advantages of the present invention will be given in part in the following description, become apparent in part from the following description, or be learned through the practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The above and / or additional aspects and advantages of the present invention will become apparent and be readily understood from the description of the embodiments in conjunction with the following drawings, where:

[0047] Figure 1 is a schematic flowchart of the confirmation method according to an embodiment of the present invention;

[0048] Figure 2 is a schematic imaging diagram according to an embodiment of the present invention;

[0049] Figure 3 is a schematic block diagram of a biological detection system according to an embodiment of the present invention;

[0050] Figure 4 is a schematic flowchart of the confirmation method according to an embodiment of the present invention;

[0051] Figure 5 is a schematic flowchart of the confirmation method according to an embodiment of the present invention;

[0052] Figure 6 is a schematic flowchart of the confirmation method according to an embodiment of the present invention;

[0053] Figure 7 is a schematic structural diagram of a biochip according to an embodiment of the present invention;

[0054] Figure 8 is a schematic flowchart of the confirmation method according to an embodiment of the present invention;

[0055] Figure 9 is a speckle image formed on the B surface according to an embodiment of the present invention;

[0056] Figure 10 is a schematic flowchart of the confirmation method according to an embodiment of the present invention;

[0057] Figure 11 is a surface image formed on the C surface according to an embodiment of the present invention;

[0058] Figure 12 is a speckle image formed on the C surface according to an embodiment of the present invention;

[0059] Figure 13 is a speckle image formed on the C surface according to an embodiment of the present invention;

[0060] Figure 14 is a schematic flowchart of the confirmation method of the embodiment of the present invention;

[0061] Figure 15 is a schematic flowchart of the confirmation method of the embodiment of the present invention;

[0062] Figure 16 is a surface image formed on the B surface of the embodiment of the present invention.

[0063] Description of reference numerals: 10, test sample; 11, biochip; 12, first base layer; 13, second base layer; 14, connection layer; 15, B surface; 16, C surface; 20, light source; 21, optical sensor; 22, objective lens; 23, collimator; 24, first filter; 25, first dichroic mirror; 26, second dichroic mirror; 27, focusing device; 28, tube lens; 29, second filter; 30, biological detection system; 31, memory; 32, processor. Detailed implementation manners

[0064] The following details the embodiments of the present invention. Examples of the embodiments are shown in the drawings, where the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the drawings are exemplary only for explaining the present invention and should not be construed as limiting the present invention.

[0065] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", etc. indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings. They are only for facilitating the description of the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention. In addition, the terms "first" and "second" are only used for descriptive purposes and should not be construed as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include one or more of the described features. In the description of the present invention, "a plurality" means two or more unless otherwise specifically defined.

[0066] In the description of the present invention, it should be noted that, unless otherwise clearly defined and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, an electrical connection, or a connection that can communicate with each other; it can be directly connected, or indirectly connected through an intermediate medium, and can be the communication inside two components or the interaction relationship between two components. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0067] In the present invention, unless otherwise clearly defined and limited, the first feature being "above" or "below" the second feature may include the direct contact between the first and second features, or may include the situation where the first and second features are not in direct contact but in contact through other features between them. Moreover, the first feature being "above", "over", and "on" the second feature includes that the first feature is directly above and obliquely above the second feature, or simply means that the horizontal height of the first feature is higher than that of the second feature. The first feature being "below", "under", and "beneath" the second feature includes that the first feature is directly below and obliquely below the second feature, or simply means that the horizontal height of the first feature is lower than that of the second feature.

[0068] The following disclosure provides many different embodiments or examples for implementing different structures of the present invention. To simplify the disclosure of the present invention, the components and settings of specific examples are described below. Of course, they are only examples and are not intended to limit the present invention. In addition, the present invention may repeat reference numerals and / or reference letters in different examples. Such repetition is for the purpose of simplification and clarity, and does not itself indicate the relationship between the various embodiments and / or settings discussed. In addition, the present invention provides examples of various specific processes and materials, but those of ordinary skill in the art can be aware of the application of other processes and / or the use of other materials.

[0069] Please refer to Figure 1 and Figure 2 , an embodiment of the present application provides a confirmation method, which is used to confirm the attribution of the optical signal of the test sample 10, and includes the following steps:

[0070] S10, controlling the light source 20 to project uneven light onto one surface of the test sample 10 to excite the test sample 10 to form an optical signal on the surface;

[0071] S20, obtaining a speckle image formed based on the optical signal through the optical sensor 21;

[0072] S30, determining the attribution of the optical signal based on the speckle image, and the attribution of the optical signal includes whether the optical signal includes non-target optical signals.

[0073] Please refer to Figure 3 , an embodiment of the present application provides a biological detection system 30, including a memory 31 and a processor 32. The processor 32 is configured to execute a computer program stored in the memory 31 to implement the method of any of the above embodiments. For example, the processor 32 is configured to control the light source 20 to project non-uniform light onto a surface of the test sample 10 to excite the test sample 10 to form an optical signal on the surface; and to obtain a speckle image formed based on the optical signal through the optical sensor 21; and to determine the attribution of the optical signal based on the speckle image.

[0074] In the confirmation method and the biological detection system 30 of the embodiment of the present application, non-uniform light is projected onto the surface of the test sample 10 to form a speckle image, and the attribution of the optical signal of the test sample 10 is confirmed through the speckle image, so as to determine whether there are defects on the surface of the test sample 10, ensure the quality of the test sample 10, and further ensure the data quality of optical detection based on the test sample.

[0075] Specifically, the optical sensor 21 is configured to collect the optical signal collected by the objective lens 22 and form an image. The optical sensor 21 is any sensor that can sense optical image information and convert it into an available output signal. The optical sensor 21 can be a area array camera for area array imaging, or a line scan camera for line scan imaging.

[0076] Speckle refers to the phenomenon that the surface of an object illuminated by a laser shows a granular structure. A speckle image refers to an image with speckles formed by laser irradiation on the surface of an object.

[0077] In one example, when the optical signal includes a non-target optical signal, it can indicate that there are defects on the surface of the test sample 10, and the surface defects include but are not limited to pits, abnormal adsorption, etc.

[0078] In some embodiments, the non-uniform light is light with non-uniform intensity and / or non-uniform distribution.

[0079] In this way, the non-uniform light irradiating the test sample 10 can form a speckle image, which is convenient for confirming the attribution of the optical signal of the test sample 10.

[0080] Specifically, the non-uniform light can be light with non-uniform intensity, or light with non-uniform distribution, or light with both non-uniform intensity and non-uniform distribution.

[0081] Please refer to Figure 4 , in some embodiments, the light source 20 includes a laser light source. Controlling the light source 20 to project non-uniform light onto a surface of the test sample 10 (step S10) includes:

[0082] S11. Control the laser light source 20 to emit self-coherent light rays so that the self-coherent light rays interfere to form non-uniform light rays.

[0083] S12. Project the non-uniform light rays onto one surface of the test sample 10.

[0084] In some embodiments, the processor 32 is used to control the laser light source 20 to emit self-coherent light rays so that the self-coherent light rays interfere to form non-uniform light rays, and to project the non-uniform light rays onto one surface of the test sample 10.

[0085] In this way, by controlling the laser light source 20 to emit self-coherent light rays, non-uniform light rays can be formed, so that a speckle image is formed on the surface of the test sample 10, which is convenient for confirming the attribution of the optical signal of the test sample 10.

[0086] Specifically, the laser light source 20 can be a solid laser source, a gas laser source, a liquid laser source, a semiconductor laser source, etc. Coherent light rays refer to two light beams with the same frequency, the same vibration direction, equal phase or a constant phase difference. The phase difference of the coherent light rays is different, forming bright spots, dark spots or between bright spots and dark spots in space.

[0087] Please refer to Figure 5 and Figure 6 , in some embodiments, based on the speckle image, determining the attribution of the optical signal (step S30) includes:

[0088] S31. When there is a bright spot in the speckle image, determine the intensity of the bright spot.

[0089] S32. Compare the intensity of the bright spot with a first preset intensity, and determine the bright spot with an intensity higher than the first preset intensity as a non-target optical signal; and / or

[0090] S33. When there is a dark spot in the speckle image, compare the intensity of the dark spot with a second preset intensity, and determine the dark spot with an intensity lower than the second preset intensity as a non-target optical signal.

[0091] In some embodiments, the processor 32 is used to compare the intensity of the bright spot with a first preset intensity when there is a bright spot in the speckle image, and determine the bright spot with an intensity higher than the first preset intensity as a non-target optical signal; and to compare the intensity of the dark spot with a second preset intensity when there is a dark spot in the speckle image, and determine the dark spot with an intensity lower than the second preset intensity as a non-target optical signal.

[0092] When uneven light irradiates a surface of the test sample 10, a fluorescence signal of the test sample 10 is excited. The excited fluorescence forms bright spots, and the unexcited fluorescence forms dark spots. When the optical signal generated on the surface of the test sample 10 is a target optical signal, the intensity of the bright spots in the speckle image is lower than or equal to a first preset intensity, and the intensity of the dark spots is higher than or equal to a second preset intensity. When there is abnormal adsorption on the surface of the test sample 10, the brightness of the excited fluorescence changes, and the intensities of the dark spots and bright spots also change. If the intensity of the bright spots is higher than the first preset intensity or the intensity of the dark spots is lower than the second preset intensity, it can be explained that the optical signal includes a non-target optical signal. Thus, by comparing the intensity of the bright spots with the first preset intensity or the intensity of the dark spots with the second preset intensity, the attribution of the optical signal can be determined.

[0093] Specifically, the first preset intensity and the second preset intensity can be set according to the intensity of the target optical signal. In some embodiments, the first preset intensity is greater than the second preset intensity.

[0094] In the embodiments of the present application, the detection sample 10 can be a test sample with a test sample to be measured on its surface, such as a biochip with a test sample to be measured on its surface; it can also be a standard sample whose surface has not yet carried a test sample, such as a biochip whose surface has not yet carried a biological molecule to be measured. At this time, the method provided by the embodiments of the present application can be used to perform quality inspection on the detection sample 10. Among them, the test sample can be a nucleic acid sample or the like.

[0095] In some embodiments, the test sample 10 includes a biochip 11, and no test sample is bound to the surface of the biochip 11.

[0096] Thus, by detecting the biochip 11 whose surface is not bound to the test sample, it can be determined whether there are defects on the surface of the biochip 11 and whether the biochip 11 is qualified.

[0097] Please refer to Figure 7 , in some embodiments, the biochip 11 includes a first base layer 12 and a second base layer 13 arranged in a stacked manner, and a connection layer 14 arranged between the first base layer 12 and the second base layer 13.

[0098] Thus, the first base layer 12 and the second base layer 13 of the biochip 11 can be detected to ensure the quality of the biochip 11, and further ensure the data quality of the optical detection based on the test sample.

[0099] Specifically, the first base layer 12 and the second base layer 13 can be solid flat substrates, and the solid substrate materials can be silicon, glass, quartz, metal, plastic, ceramic, rubber, gel, paper, nylon, etc. When the first base layer 12 and the second base layer 13 are glass parallel substrates, the connecting layer 14 can be a parallel micro-component with a diameter of 100 um, spaced 20 mm apart and coated with silica gel on the surface. The connecting layer 14 is formed with multiple micro-channel channels, and the sample to be tested can be accommodated in the channels.

[0100] In some embodiments, the first base layer 12 is a substrate layer with a light transmittance greater than or equal to 90%.

[0101] In this way, the first base layer 12 has good light transmittance, so that non-uniform light can enter the chip, and the optical signals in the chip can also be emitted to the outside, and the speckle image imaging effect is better.

[0102] Specifically, the light transmittance of the first base layer 12 can be 90%, 91%, 92%, 93%, 94%, 95%, etc., and the material of the first base layer 12 can be glass, quartz, etc.

[0103] Please refer to Figure 8 , in some embodiments, controlling the light source 20 to project non-uniform light onto a surface of the test sample 10 (step S10) includes:

[0104] S13, controlling the light source 20 to project non-uniform light onto the surface where the first base layer 12 of the biochip 11 is located.

[0105] In some embodiments, the processor 32 is used to control the light source 20 to project non-uniform light onto the surface where the first base layer 12 of the biochip 11 is located.

[0106] In this way, non-uniform light can be projected onto the first base layer 12 in a critical illumination manner to form a speckle image, which is convenient for detecting the surface of the first base layer 12.

[0107] Specifically, the light source 20 can project non-uniform light onto the surface where the first base layer 12 is located in a critical illumination manner to detect the surface of the first base layer 12, or can project non-uniform light onto the surface where the second base layer 13 is located in a critical illumination manner to detect the surface of the second base layer 13.

[0108] Please refer to Figure 8 , in some embodiments, the first base layer 12 includes a B surface 15 facing the second base layer 13, and obtaining a speckle image formed based on the optical signal through the optical sensor 21 (step S20) includes:

[0109] S21. Obtain a first speckle image formed by an optical signal generated based on the B surface 15 through the optical sensor 21.

[0110] In some embodiments, the processor 32 is configured to obtain a first speckle image formed by an optical signal generated based on the B surface 15 through the optical sensor 21.

[0111] Thus, the B surface 15 of the first base layer 12 can be detected through the first speckle image.

[0112] Specifically, the light source 20 projects inhomogeneous light onto the B surface 15, exciting the B surface 15 to generate an optical signal, and the optical signal converges to the optical sensor 21 to form a first speckle image.

[0113] Please refer to Figure 8 , in some embodiments, based on the speckle image, determine the attribution of the optical signal, where the attribution of the optical signal includes whether the optical signal includes a non-target optical signal (step S30), including:

[0114] S34. When there are dark spots in the first speckle image, compare the intensity of the dark spots with a second preset intensity, and determine the dark spots with an intensity lower than the second preset intensity as non-target optical signals.

[0115] In some embodiments, the processor 32 is configured to, when there are dark spots in the first speckle image, compare the intensity of the dark spots with a second preset intensity, and determine the dark spots with an intensity lower than the second preset intensity as non-target optical signals. It should be understood that the second preset intensity can be determined based on the intensity obtained when performing the above operations on the biochip 11 without surface defects.

[0116] When there is no abnormal adsorption on the B surface 15, the fluorescence signal is excited by inhomogeneous light to form speckles, and the first speckle image is arranged in a speckle characteristic pattern; when there is abnormal adsorption on the B surface 15, the fluorescence signal in the abnormal adsorption area is not excited and forms dark spots with lower intensity. If the intensity of the dark spots is lower than the second preset intensity, it can indicate that the fluorescence brightness excited by the B surface 15 decreases and the number of fluorescence excited by the B surface 15 decreases, that is, there is abnormal adsorption on the B surface 15. Thus, by comparing the intensity of the dark spots with the second preset intensity, the attribution of the optical signal can be determined, thereby detecting the B surface 15 of the first base layer 12 of the biochip 11.

[0117] Combined with Figure 9 , Figure 9 is an image with speckle characteristics formed for the B surface 15, Figure 9 in which the dark spots and bright spots are unevenly distributed, and the shooting effect is similar to that of a Köhler illumination system with laser speckles. According to Figure 9 it can be determined that there are no defects on the B surface 15.

[0118] Please refer toFigure 10 , in some embodiments, the second base layer 13 includes a C surface 16 facing the first base layer 12. Obtaining a speckle image formed based on an optical signal through the optical sensor 21 (step S20) includes:

[0119] S22, obtaining a second speckle image formed based on the optical signal generated by the C surface 16 through the optical sensor 21.

[0120] In some embodiments, the processor 32 is configured to obtain a second speckle image formed based on the optical signal generated by the C surface 16 through the optical sensor 21.

[0121] Thus, the C surface 16 of the second base layer 13 can be detected through the second speckle image.

[0122] Specifically, the light source 20 projects inhomogeneous light onto the C surface 16, exciting the C surface 16 to generate an optical signal, and the optical signal converges to the optical sensor 21 to form a second speckle image.

[0123] Please refer to Figure 10 , in some embodiments, based on the speckle image, determining the attribution of the optical signal, where the attribution of the optical signal includes whether the optical signal includes a non-target optical signal (step S30), includes:

[0124] S35, dividing the second speckle image into multiple regions;

[0125] S36, if the average intensity of the region pixels is higher than a first preset intensity, determining that the optical signal of the C surface 16 has a non-target optical signal.

[0126] In some embodiments, the processor 32 is configured to divide the second speckle image into multiple regions; and to determine that the optical signal of the C surface 16 has a non-target optical signal if the average intensity of the region pixels is higher than a first preset intensity.

[0127] When there is no abnormal adsorption on the C surface 16, the second speckle image is an image of background light reflection; when there is abnormal adsorption on the C surface 16, the adsorption amount of each region is different, so the excited fluorescence brightness is different, and the abnormal adsorption region forms a speckle, and the intensity of the speckle is higher than the first preset intensity. Thus, by comparing the average intensity of the region pixels of the second speckle image with the first preset intensity, the attribution of the optical signal can be determined, thereby detecting the C surface 16 of the second base layer 13 of the biochip 11.

[0128] Specifically, the second speckle image can be divided into multiple regions such as three, four, five, etc. Combining Figures 11 - 13 , Figure 11 an image without speckle features formed for the C surface 16, Figure 12 andFigure 13 An image with speckle features formed on the surface C 16 Figure 11 In [reference], the light of abnormal adsorption and background reflection is easily confused, and it is impossible to judge the problem of abnormal adsorption on the surface C 16. However, Figure 12 and Figure 13 In [reference], the bright spots are arranged in a speckle pattern. According to the bright spots in Figure 12 the edge adsorption problem of the surface C 16 can be judged according to the bright spots in Figure 13 the central adsorption problem of the surface C 16 can be judged according to the bright spots in

[0129] In some embodiments, the optical signal is a fluorescence signal.

[0130] Thus, when the uneven light emitted by the light source 20 irradiates the surface of the test sample 10, the fluorescence signal will emit light, and the emitted light will converge to the optical sensor 21 to form a speckle image.

[0131] Specifically, in some embodiments, the surface B 15 is provided with a fluorescence signal. When the light irradiates the surface B 15, the fluorescence signal will emit light. When there are dark spots with intensity lower than the second preset intensity in the first speckle image, it means that there is no fluorescence signal at the position corresponding to the speckle on the surface B 15, that is, there is a defect at the position corresponding to the speckle on the surface B 15. The surface C 16 has no fluorescence signal. When the light irradiates the surface C 16, when there are bright spots with intensity higher than the first preset intensity in the second speckle image, it means that there is a fluorescence signal at the position corresponding to the bright spot on the surface C 16, that is, there is a defect at the position corresponding to the bright spot on the surface C 16.

[0132] Please refer to Figure 14 , in some embodiments, the method includes:

[0133] S40, a light homogenizing device is arranged on the optical path of the light source 20 to homogenize the light emitted by the light source 20, so as to project uniform light onto one surface of the test sample 10 to excite the test sample 10 to form an optical signal on the surface;

[0134] S50, obtaining a surface image formed based on the optical signal through the optical sensor 21;

[0135] S60, based on the surface image, determining the attribution of the optical signal, and the attribution of the optical signal includes whether the optical signal includes non-target optical signals.

[0136] In some embodiments, the processor 32 is configured to set a light homogenizing device on the optical path of the light source 20 to homogenize the light emitted by the light source 20, so as to project uniform light onto one surface of the test sample 10 to excite the test sample 10 to form an optical signal on the surface; and is configured to obtain a surface image formed based on the optical signal through the optical sensor 21; and is configured to determine the attribution of the optical signal based on the surface image, where the attribution of the optical signal includes whether the optical signal includes a non-target optical signal.

[0137] In this way, by projecting uniform light onto the surface of the test sample 10 to form a surface image, and confirming the attribution of the optical signal of the test sample 10 through the surface image, it can be determined whether there are defects on the surface of the test sample 10, ensuring the quality of the test sample 10, and further ensuring the data quality of the optical detection based on the test sample.

[0138] Specifically, the light homogenizing device can be a light homogenizing sheet. The light homogenizing device can convert the light emitted by the light source 20 into a light beam with an arbitrary shape and a uniform intensity distribution. The light homogenizing device can selectively cut into or out of the optical path of the light source 20. When forming a speckle image, the light homogenizing device is in the cut-out state; when forming a surface image, the light homogenizing device is in the cut-in state.

[0139] Please refer to Figure 15 , in some embodiments, determining the attribution of the optical signal based on the surface image (step S60) includes:

[0140] S61, when there are dark spots in the surface image, comparing the intensity of the dark spots with a second preset intensity, and determining the dark spots with an intensity lower than the second preset intensity as non-target optical signals.

[0141] In some embodiments, when there are dark spots in the surface image, the processor 32 is configured to compare the intensity of the dark spots with a second preset intensity, and determine the dark spots with an intensity lower than the second preset intensity as non-target optical signals.

[0142] When there is no abnormal adsorption on the B surface 15 of the test sample 10, the intensity of the surface image is the fluorescence brightness excited. When there is abnormal adsorption on the B surface 15 of the test sample 10, the fluorescence signal in the abnormal adsorption area is not excited and forms a dark spot. If the intensity of the dark spot is lower than the second preset intensity, it can indicate that the optical signal generated on the B surface 15 of the test sample 10 includes non-target optical signals. In this way, by comparing the intensity of the dark spot with the second preset intensity, the attribution of the optical signal can be determined, thereby detecting the surface of the test sample 10.

[0143] Combined with Figure 16 , Figure 16 is an image without speckle features formed on the B surface 15. According to Figure 16 the dark spots in it, the defects existing on the B surface 15 can be judged.

[0144] Please refer to Figure 2 , in some embodiments, an objective lens 22 and a collimator 23 are disposed between the test sample 10 and the light source 20. The collimator 23 is configured to collimate the light emitted by the light source 20 into parallel light; the objective lens 22 is configured to converge the parallel light onto the surface of the test sample 10 to form critical illumination on the surface of the test sample 10.

[0145] In this way, the collimator 23 can collimate the non-uniform light from the light source 20 into parallel light, so that the non-uniform light remains parallel during transmission, reducing energy loss and beam divergence. Critical illumination converges the parallel light onto the surface of the test sample 10, which can reduce the interference of the untested surface on the test surface and improve the imaging effect.

[0146] Specifically, the objective lens 22 is configured to collect the optical signals from the B surface 15 and the C surface 16. The objective lens 22 can be an off-the-shelf objective lens, such as an infinity conjugate objective lens, or the objective lens 22 can also be a customized objective lens. The focal length of the objective lens 22 can be set according to the imaging requirements. The optical axis of the objective lens 22 is perpendicular to the B surface 15 and the C surface 16. The collimator 23 can be a spherical lens or an aspherical lens, and the collimator 23 can be a single lens or a lens group composed of multiple lenses.

[0147] In some embodiments, critical illumination adopts epi-illumination critical illumination, that is, the incident direction of the parallel light is the same as the incident light direction of the objective lens 22. When the critical illumination irradiates the parallel light onto the B surface 15, it can reduce the interference of the C surface 16 on the imaging of the B surface 15. Similarly, when the critical illumination irradiates the parallel light onto the C surface 16, it can reduce the interference of the B surface 15 on the imaging of the C surface 16.

[0148] Please refer to Figure 2 , in some embodiments, a first dichroic mirror 25 is disposed between the collimator 23 and the objective lens 22. The first dichroic mirror 25 is disposed on the optical axis of the objective lens 22. The first dichroic mirror 25 is configured to reflect the parallel light to the objective lens 22 and transmit the optical signal from the surface of the test sample 10 to the optical sensor 21.

[0149] In this way, the first dichroic mirror 25 reflects the parallel light to the objective lens 22, so that the parallel light irradiates on the test sample 10 after passing through the objective lens 22 to excite the surface of the test sample 10 to form an optical signal.

[0150] In some embodiments, the first dichroic mirror 25 is placed at an angle of 45° with the optical axis of the objective lens 22, and the objective lens 22 and the light source 20 are located on the same side of the first dichroic mirror 25, so that the parallel light is incident on the objective lens 22.

[0151] A first filter 24 is disposed between the collimating mirror 23 and the first dichroic mirror 25. The first filter 24 is configured to perform band-pass filtering on the parallel light from the collimating mirror 23 to remove stray light in the light beam, which is beneficial to further improve the excitation efficiency and imaging quality.

[0152] Please refer to Figure 2 , in some embodiments, a second dichroic mirror 26 is disposed on a side of the first dichroic mirror 25 away from the objective lens 22. A focusing device 27 is disposed on a side of the second dichroic mirror 26 deviating from the optical axis of the objective lens 22. The focusing device 27 is configured to emit a focusing light beam. The focusing light beam is reflected by the second dichroic mirror 26 and propagates along the optical axis of the objective lens 22 and irradiates the test sample 10. The focusing device 27 is further configured to receive the focusing light beam reflected from the test sample 10 and reflected by the second dichroic mirror 26; the second dichroic mirror 26 is further configured to transmit the optical signal from the first dichroic mirror 25 to the optical sensor 21.

[0153] Thus, the focusing device 27 is mainly configured to mark the object distance after the objective lens 22 is focused, and to monitor the change amount of the object distance in real time during the optical detection based on the test sample, and correct it to ensure that the system is always in clear focus, ensure the accuracy of the image collected by the optical sensor 21, and finally obtain a clear image within the depth of focus range.

[0154] In some embodiments, the second dichroic mirror 26 is placed at an angle of 45° with the optical axis of the objective lens 22, and the objective lens 22 and the focusing device 27 are located on the same side of the second dichroic mirror 26, so that the focusing light beam is incident on the objective lens 22.

[0155] A tube lens 28 is disposed between the second dichroic mirror 26 and the optical sensor 21. The tube lens 28 is configured to cooperate with the objective lens 22 to converge the optical signal from the surface of the test sample 10 to the optical sensor 21 for imaging. A second filter 29 is disposed on a side of the tube lens 28 facing away from the optical sensor 21. The second filter 29 can filter light, allow the fluorescence band to pass through, and cut off the unnecessary laser band, thereby improving the imaging quality of the optical sensor 21.

[0156] In some embodiments, the light source 20 is a multimode laser.

[0157] Thus, the laser emitted by the multimode laser irradiates the test sample 10 to generate speckles, which is convenient for forming a speckle image.

[0158] Specifically, a multimode laser refers to a laser that simultaneously generates two or more modes of emitted light.

[0159] The confirmation method provided by the embodiments of the present application can be used to confirm the attribution of the optical signal of the test sample, and is applicable to the detection and quality control of chips used in gene sequencing, nucleic acid fragment analyzers, etc. The embodiments of the present application provide a non-volatile computer-readable storage medium storing a computer program, and when the computer program is executed by one or more processors 32, the method of any of the above embodiments is implemented.

[0160] Specifically, in one embodiment, the processor 32 may be a central processing unit (CPU). The processor 32 may also be other general-purpose processors, digital signal processors (DSPs), application specific integrated circuits (ASICs), field-programmable gate arrays (FPGAs) or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc. chips, or combinations of the above types of chips.

[0161] The computer program may be stored in the memory 31. The memory 31, as a non-transitory computer-readable storage medium, can be used to store non-transitory software programs, non-transitory computer-executable programs, and modules, such as the program instructions / modules corresponding to the methods in the above method embodiments. The processor 32 executes various functional applications and data processing of the processor 32 by running the non-transitory software programs, instructions, and modules stored in the memory 31, that is, the control method in the above method embodiments is implemented.

[0162] The storage medium may include, but is not limited to: USB flash drives, read-only memories (ROMs), random access memories (RAMs), mobile hard disks, magnetic disks, or optical discs, etc., various media that can store computer programs.

[0163] In the description of this specification, the descriptions with reference to terms such as "one embodiment", "certain embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples", etc. mean that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic descriptions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments or examples.

[0164] Although embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the claims and their equivalents.

Claims

1. A confirmation method, characterized in that, The method is used to confirm the attribution of the optical signal of a test sample, and includes the following steps: Controlling a light source to project inhomogeneous light onto a surface of the test sample to excite the test sample to form an optical signal on the surface; Obtaining a speckle image formed based on the optical signal through an optical sensor; Based on the speckle image, determining the attribution of the optical signal, where the attribution of the optical signal includes whether the optical signal includes non-target optical signals; 2. The method according to claim 1, characterized in that, The inhomogeneous light is light with inhomogeneous intensity and / or inhomogeneous distribution; Optionally, the light source includes a laser light source, and controlling the light source to project inhomogeneous light onto a surface of the test sample includes: Controlling the laser light source to emit self-coherent light so that the self-coherent light interferes to form the inhomogeneous light; Projecting the inhomogeneous light onto a surface of the test sample; 3. The method according to claim 1, wherein The determining the attribution of the optical signal based on the speckle image includes: When there is a bright spot in the speckle image, determining the intensity of the bright spot; Comparing the intensity of the bright spot with a first preset intensity, and determining the bright spot with an intensity higher than the first preset intensity as a non-target optical signal; and / or When there is a dark spot in the speckle image, comparing the intensity of the dark spot with a second preset intensity, and determining the dark spot with an intensity lower than the second preset intensity as a non-target optical signal; 4. The method according to claim 3, characterized in that, The test sample includes a biochip, and no sample to be detected is bound to the surface of the biochip; Optionally, the biochip includes a first base layer and a second base layer arranged in a stacked manner, and a connection layer arranged between the first base layer and the second base layer; Optionally, the first base layer is a substrate layer with a light transmittance greater than or equal to 90%; 5. The method according to claim 4, characterized in that, The controlling the light source to project inhomogeneous light onto a surface of the test sample includes: Controlling the light source to project inhomogeneous light onto the surface of the biochip where the first base layer is located; Optionally, the first base layer includes a B surface facing the second base layer; The obtaining a speckle image formed based on the optical signal through an optical sensor includes: Obtaining a first speckle image formed based on the optical signal generated on the B surface through the optical sensor; Optionally, the determining the attribution of the optical signal based on the speckle image, where the attribution of the optical signal includes whether the optical signal includes non-target optical signals, includes: When there is a dark spot in the first speckle image, comparing the intensity of the dark spot with a second preset intensity, and determining the dark spot with an intensity lower than the second preset intensity as a non-target optical signal; 6. The method according to claim 4 or 5, characterized in that The second base layer includes a C surface facing the first base layer; The obtaining a speckle image formed based on the optical signal through an optical sensor includes: Obtaining a second speckle image formed based on the optical signal generated on the C surface through the optical sensor; Optionally, the determining the attribution of the optical signal based on the speckle image, where the attribution of the optical signal includes whether the optical signal includes non-target optical signals, includes: Dividing the second speckle image into multiple regions; If the average intensity of the pixels in the region is higher than the first preset intensity, it is determined that there is a non-target optical signal in the optical signal of the C surface; Optionally, the optical signal is a fluorescence signal.

7. The method according to any one of claims 1 to 6, characterized in that, The method includes: A light homogenizing device is arranged on the optical path of the light source to homogenize the light emitted by the light source, so as to project uniform light onto a surface of the test sample to excite the test sample to form an optical signal on the surface; An optical sensor is used to obtain a surface image formed based on the optical signal; Based on the surface image, the attribution of the optical signal is determined, and the attribution of the optical signal includes whether the optical signal includes a non-target optical signal; Optionally, determining the attribution of the optical signal based on the surface image includes: When there is a dark spot in the surface image, the intensity of the dark spot is compared with a second preset intensity, and the dark spot with an intensity lower than the second preset intensity is determined as a non-target optical signal.

8. The method according to any one of claims 1 to 7, characterized in that An objective lens and a collimating mirror are arranged between the test sample and the light source, and the collimating mirror is used to collimate the light emitted by the light source into parallel light; The objective lens is used to converge the parallel light onto the surface of the test sample to form critical illumination on the surface of the test sample; Optionally, a first dichroic mirror is arranged between the collimating mirror and the objective lens. The first dichroic mirror is arranged on the optical axis of the objective lens. The first dichroic mirror is used to reflect the parallel light to the objective lens and to transmit the optical signal from the surface of the test sample to the optical sensor; Optionally, a second dichroic mirror is arranged on the side of the first dichroic mirror away from the objective lens, and a focusing device is arranged on the side of the second dichroic mirror deviating from the optical axis of the objective lens. The focusing device is used to emit a focusing beam. The focusing beam is reflected by the second dichroic mirror and propagates along the optical axis of the objective lens and irradiates the test sample. The focusing device is also used to receive the focusing beam reflected from the test sample and reflected by the second dichroic mirror; The second dichroic mirror is also used to transmit the optical signal from the first dichroic mirror to the optical sensor; Optionally, the light source is a multimode laser.

9. A biological detection system, characterized in that, It includes a memory and a processor. The processor is used to execute the computer program stored in the memory to implement the method according to any one of claims 1-8.

10. A non-volatile computer-readable storage medium storing a computer program, characterized in that, When the computer program is executed by one or more processors, the method according to any one of claims 1-8 is implemented.

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