Fluorescence value compensation method, device, equipment and storage medium for solid standard

Abstract

The present invention proposes a fluorescence value compensation method, device, equipment and storage medium for a solid-state standard, and belongs to the field of SiPM optical measurement. The method comprises: arranging a first solid-state standard and a second solid-state standard in an optical detection device, scanning them at a calibration temperature, a reference temperature and an actual measurement stage respectively, constructing a first mapping relationship through the fluorescence values ​​of the two solid-state standards measured at the calibration temperature and the reference temperature, utilizing the characteristic that the two solid-state standards are not linearly correlated with temperature changes, compensating the fluorescence value of the first solid-state standard through the second solid-state standard, and making the fluorescence value of the first solid-state standard at any temperature equivalent to the fluorescence value at the reference temperature, so that the solid-state standard is affected by temperature fluctuations close to that of a liquid standard, thereby improving the accuracy of temperature compensation and optical detection.

Description

Technical Field

[0001] The present invention relates to the field of optical detection technology based on SiPM, and in particular to a fluorescence value compensation method, device, equipment and storage medium for a solid-state standard. Background Art

[0002] Silicon photomultiplier (SiPM) is a new type of photoelectric detection device with the characteristics of high gain, high sensitivity, low bias voltage, insensitivity to magnetic field, compact structure, etc., and can be applied to the field of extremely weak light detection. In the prior art with application number CN202410027463.0, a temperature compensation method based on SiPM is disclosed, which uses solid-state standard as a reference for temperature compensation to eliminate the detection value offset caused by the working temperature of SiPM components to a certain extent.

[0003] Compared with liquid standards, solid standards have obvious advantages in manufacturing, storage, transportation and use. When the sample is in a constant temperature detection environment, such as loop-mediated isothermal amplification (LAMP) detection or recombinase polymerase amplification (RPA) detection, the use of solid standards can effectively compensate for the drift of SiPM components caused by changes in ambient temperature.

[0004] However, some optical tests use liquid reagents as the test samples, such as fluorescent polymerase chain reaction (PCR) test. When the test samples are in a constantly changing temperature environment, since the fluorescence value of solid standards is usually more affected by temperature changes than liquid reagents, if solid standards are used directly to perform temperature compensation on SiPM components in this scenario, the accuracy of temperature compensation will be affected, which in turn affects the accuracy of optical testing. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention proposes a fluorescence value compensation method, device, equipment, and storage medium for a solid standard, which can compensate the fluorescence value of a first solid standard obtained at any temperature to a mapping value at a reference temperature, so that the solid standard is affected by temperature fluctuations close to that of a liquid standard, thereby improving the accuracy of temperature compensation and optical detection.

[0006] In a first aspect, an embodiment of the present invention provides a fluorescence value compensation method for a solid-state standard, which is applied to an optical detection device, wherein the optical detection device includes a SiPM optical component and a detection unit, wherein the detection unit is provided with adjacent first solid-state standard and second solid-state standard, and the fluorescence values ​​of the first solid-state standard and the second solid-state standard are not linearly correlated with temperature changes, and the fluorescence value compensation method of the solid-state standard includes:

[0007] At a calibration temperature, performing N scanning operations through the SiPM optical component to obtain N groups of calibration data sets, wherein the calibration data sets include a first calibration fluorescence value and a second calibration fluorescence value obtained based on the same scan, the first calibration fluorescence value corresponds to the first solid-state standard, the second calibration fluorescence value corresponds to the second solid-state standard, and N is a positive integer;

[0008] At a preset reference temperature, performing a scanning operation through the SiPM optical component to obtain a first reference fluorescence value and a second reference fluorescence value, wherein the first reference fluorescence value corresponds to the first solid-state standard, and the second reference fluorescence value corresponds to the second solid-state standard;

[0009] Determine the ratio of the first calibration fluorescence value and the second calibration fluorescence value of the same group as the first fluorescence ratio of the calibration data group, and construct a first mapping relationship based on the first calibration fluorescence value, the first reference fluorescence value and the first fluorescence ratio, wherein the first mapping relationship is used to characterize the mapping ratio of the first calibration fluorescence value and the first reference fluorescence value;

[0010] After starting the sample detection, a scanning operation is performed through the SiPM optical component to obtain a first measured fluorescence value of the first solid-state standard and a second measured fluorescence value of the second solid-state standard, and a first compensation fluorescence value is determined based on the first measured fluorescence value, the second measured fluorescence value and the first mapping relationship, wherein the first compensation fluorescence value is used to characterize the mapping value of the first measured fluorescence value at the reference temperature.

[0011] According to some embodiments of the present invention, the SiPM optical component is provided with a first light source, and the scanning operation includes:

[0012] Turning off the first light source, scanning the first solid standard object based on the SiPM optical component to obtain a system dark background value;

[0013] Turning on the first light source, scanning the first solid standard object based on the SiPM optical component to obtain a first detection fluorescence value, and scanning the second solid standard object to obtain a second detection fluorescence value;

[0014] Obtain a first target fluorescence value based on the first detected fluorescence value and the system dark background value, and obtain a second target fluorescence value based on the second detected fluorescence value and the system dark background value;

[0015] Wherein, when the scanning operation is performed at the calibration temperature, the first target fluorescence value is the first calibration fluorescence value, and the second target fluorescence value is the second calibration fluorescence value; or, when the scanning operation is performed at the reference temperature, the first target fluorescence value is the first reference fluorescence value, and the second target fluorescence value is the second reference fluorescence value; or, when the scanning operation is performed in the actual measurement stage, the first target fluorescence value is the first measured fluorescence value, and the second target fluorescence value is the second measured fluorescence value.

[0016] According to some embodiments of the present invention, constructing a first mapping relationship based on the first calibrated fluorescence value, the first reference fluorescence value and the first fluorescence ratio (or constructing a second mapping relationship based on the second calibrated fluorescence value, the second reference fluorescence value and the second fluorescence ratio) includes:

[0017] Determining a first light intensity ratio based on the first calibration fluorescence value and the first reference fluorescence value, wherein the first light intensity ratio is used to indicate a ratio of a first calibration optical intensity to a first reference optical intensity, the first calibration optical intensity being the emission light intensity of the first solid standard at the calibration temperature, and the first reference optical intensity being the emission light intensity of the first solid standard at the reference temperature;

[0018] The first mapping relationship is constructed according to the first light intensity ratio and the first fluorescence ratio.

[0019] According to some embodiments of the present invention, constructing the first mapping relationship according to the first light intensity ratio and the first fluorescence ratio includes:

[0020] constructing a first mapping relationship based on the first light intensity ratio and the first fluorescence ratio, wherein the first mapping relationship includes a plurality of fitting coefficients;

[0021] Based on the first reference fluorescence value, the N first calibration fluorescence values ​​and the N first fluorescence ratios, a fitting calculation is performed on the first mapping relationship to determine the value of each of the fitting coefficients.

[0022] According to some embodiments of the present invention, the expression of the first mapping relationship is:

[0023] ;in, is the first calibration optical intensity, is the first reference optical intensity, is the calibration temperature, is the reference temperature, for indicating the first solid standard, { , , , ···} is the fitting coefficient, x is the first fluorescence ratio, and x , is the first calibrated fluorescence value, is the second calibrated fluorescence value, is the first light intensity ratio.

[0024] According to some embodiments of the present invention, determining a first compensated fluorescence value based on the first measured fluorescence value, the second measured fluorescence value and the first mapping relationship includes:

[0025] determining a second fluorescence ratio based on the first measured fluorescence value and the second measured fluorescence value;

[0026] replacing the first fluorescence ratio in the first mapping relationship with the second fluorescence ratio, and calculating a value of the first light intensity ratio;

[0027] The first compensated fluorescence value is determined based on the first measured fluorescence value and the first light intensity ratio.

[0028] According to some embodiments of the present invention, after performing a scanning operation by the SiPM optical component to obtain a first reference fluorescence value and a second reference fluorescence value, the method further includes:

[0029] Determine the ratio of the second calibration fluorescence value to the first calibration fluorescence value of the same group as the second fluorescence ratio of the calibration data group, and construct a second mapping relationship based on the second calibration fluorescence value, the second reference fluorescence value and the second fluorescence ratio, wherein the second mapping relationship is used to characterize the mapping ratio of the second calibration fluorescence value to the second reference fluorescence value;

[0030] After starting sample detection, a first measured fluorescence value of the first solid-state standard and a second measured fluorescence value of the second solid-state standard are obtained through the SiPM optical component, and a second compensated fluorescence value is determined based on the first measured fluorescence value, the second measured fluorescence value and the second mapping relationship, wherein the second compensated fluorescence value is used to characterize the mapping value of the second measured fluorescence value at the reference temperature.

[0031] In a second aspect, an embodiment of the present invention provides a fluorescence value compensation device for a solid-state standard, comprising at least one control processor and a memory for communicating with the at least one control processor; the memory stores instructions that can be executed by the at least one control processor, and the instructions are executed by the at least one control processor so that the at least one control processor can execute the fluorescence value compensation method for a solid-state standard as described in the first aspect above.

[0032] In a third aspect, an embodiment of the present invention provides an electronic device, comprising the fluorescence value compensation device of the solid-state standard as described in the second aspect above.

[0033] In a fourth aspect, an embodiment of the present invention provides a computer-readable storage medium storing computer-executable instructions, wherein the computer-executable instructions are used to execute the fluorescence value compensation method of a solid-state standard as described in the first aspect above.

[0034] The fluorescence value compensation method for a solid standard according to an embodiment of the present invention has at least the following beneficial effects: at a calibration temperature, N scanning operations are performed by the SiPM optical component to obtain N groups of first calibration fluorescence values ​​and second calibration fluorescence values; at a preset reference temperature, a scanning operation is performed by the SiPM optical component to obtain a first reference fluorescence value and a second reference fluorescence value, wherein the first reference fluorescence value corresponds to the first solid standard, and the second reference fluorescence value corresponds to the second solid standard; the ratio of the first calibration fluorescence value and the second calibration fluorescence value of the same group is determined as the first fluorescence ratio of the calibration data group, and a first mapping relationship is constructed based on the first calibration fluorescence value, the first reference fluorescence value and the first fluorescence ratio, wherein the first mapping relationship is used to characterize the mapping ratio of the first calibration fluorescence value and the first reference fluorescence value; after starting sample detection, a scanning operation is performed by the SiPM optical component to obtain a first measured fluorescence value of the first solid standard and a second measured fluorescence value of the second solid standard, and a first compensation fluorescence value is determined based on the first measured fluorescence value, the second measured fluorescence value and the first mapping relationship, wherein the first compensation fluorescence value is used to characterize the mapping value of the first measured fluorescence value at the reference temperature. According to the technical solution of the embodiment of the present invention, it is possible to utilize the characteristic that the two solid-state standards are non-linearly correlated with temperature changes, and use the second solid-state standard to compensate the fluorescence value of the first solid-state standard, so that the fluorescence value of the first solid-state standard at any temperature is equivalent to the fluorescence value at the reference temperature, so that the solid-state standard is affected by temperature fluctuations close to that of the liquid standard, thereby improving the accuracy of temperature compensation and optical detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1is a front view of an optical detection device provided by an embodiment of the present invention;

[0036] Figure 2 is a flow chart of a fluorescence value compensation method for a solid standard provided by another embodiment of the present invention;

[0037] Figure 3 It is a schematic diagram of temperature compensation by liquid standard and solid standard in the prior art;

[0038] Figure 4 is a schematic diagram of temperature compensation by two solid-state standards provided by another embodiment of the present invention;

[0039] Figure 5 is a curve schematic diagram of a first light intensity ratio provided by another embodiment of the present invention;

[0040] Figure 6 It is a structural diagram of a fluorescence value compensation device for a solid-state standard provided in another embodiment of the present invention. DETAILED DESCRIPTION

[0041] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0042] In the description of the present invention, it should be understood that descriptions involving orientations, such as up, down, front, back, left, right, etc., and orientations or positional relationships indicated are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0043] In the description of the present invention, "several" means one or more, "more" means more than two, "greater than", "less than", "exceed" etc. are understood as not including the number itself, and "above", "below", "within" etc. are understood as including the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing the technical features, and cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0044] In the description of the present invention, unless otherwise clearly defined, terms such as setting, installing, connecting, etc. should be understood in a broad sense, and technicians in the relevant technical field can reasonably determine the specific meanings of the above terms in the present invention based on the specific content of the technical solution.

[0045] The embodiments of the present invention provide a fluorescence value compensation method, device, equipment, and storage medium for a solid-state standard, wherein the fluorescence value compensation method for a solid-state standard comprises: performing N scanning operations at a calibration temperature through the SiPM optical component to obtain N groups of first calibration fluorescence values ​​and second calibration fluorescence values; performing a scanning operation at a preset reference temperature through the SiPM optical component to obtain a first reference fluorescence value and a second reference fluorescence value, wherein the first reference fluorescence value corresponds to the first solid-state standard, and the second reference fluorescence value corresponds to the second solid-state standard; determining the ratio of the first calibration fluorescence value to the second calibration fluorescence value of the same group as the calibration data group; A first fluorescence ratio, a first mapping relationship is constructed based on the first calibration fluorescence value, the first reference fluorescence value and the first fluorescence ratio, wherein the first mapping relationship is used to characterize the mapping ratio of the first calibration fluorescence value and the first reference fluorescence value; after starting the sample detection, a scanning operation is performed through the SiPM optical component to obtain the first measured fluorescence value of the first solid standard and the second measured fluorescence value of the second solid standard, and a first compensation fluorescence value is determined based on the first measured fluorescence value, the second measured fluorescence value and the first mapping relationship, wherein the first compensation fluorescence value is used to characterize the mapping value of the first measured fluorescence value at the reference temperature. According to the technical solution of the embodiment of the present invention, it is possible to utilize the characteristic that the two solid standards are not linearly correlated with temperature changes, and the fluorescence value of the first solid standard is compensated by the second solid standard, and the fluorescence value of the first solid standard at any temperature is equivalent to the fluorescence value at the reference temperature, so that the solid standard is affected by temperature fluctuations close to the liquid standard, thereby improving the accuracy of temperature compensation and optical detection.

[0046] First, refer to Figure 1 , Figure 1 This is a front view of an optical detection device provided in an embodiment of the present invention. The optical detection device of this embodiment is applied to an optical detection device. The optical detection device includes a SiPM optical component 20 and a detection unit 10. The detection unit 10 is provided with adjacent first solid-state standard objects 11 and second solid-state standard objects 12. The fluorescence values ​​of the first solid-state standard objects 11 and the second solid-state standard objects 12 are not linearly correlated with temperature changes.

[0047] It should be noted that the specific operating principle of the optical detection equipment is disclosed in the prior art. For example, the detection part 10 is also provided with a plurality of sample channels 13 for setting the samples to be tested, and each sample channel 13 is provided with SiPM components and channel light sources. The SiPM optical component moves horizontally along the track to scan different channels and obtain relevant detection values. The specific operating principle of the optical detection equipment is not elaborated in this embodiment.

[0048] It should be noted that in the prior art, the detection unit 10 has only one solid standard. When the sample to be tested is a liquid substance and is in a constantly changing temperature environment, the fluorescence values ​​of the solid substance and the liquid substance are affected differently by temperature changes, which makes the result fluctuate when the solid standard is used to perform temperature compensation on the SiPM. In actual measurement, the fluorescence value of the solid standard varies with temperature by about 0.1% to 0.2% / °C, while the gain of the SiPM varies with temperature by about 3% to 10% / °C. The fluorescence value of the solid standard varies with temperature less than the gain of the SiPM. If only one solid standard is used for temperature compensation according to the structure of the prior art, the stability and accuracy of the temperature compensation will be affected.

[0049] like Figure 3 As shown, Figure 3 It is a schematic diagram of using the existing technology to carry out temperature compensation for the first sample, the second sample and the third sample respectively by carrying solid standard materials and liquid standard materials when the temperature variation range is large. Figure 3 The vertical axis is the fluorescence intensity (FL Intensity), and the horizontal axis is the operating temperature of the SiPM component. The curve of the first sample before temperature compensation is the first pre-compensation curve 113, the curve obtained by temperature compensation through the liquid standard is the first liquid compensation curve 112, and the curve obtained by temperature compensation through the solid standard is the first solid compensation curve 111; similarly, there are the second pre-compensation curve 213, the second liquid compensation curve 212, and the second solid compensation curve 211 of the second sample, and the third pre-compensation curve 313, the third liquid compensation curve 312, and the third solid compensation curve 311 of the third sample. Figure 3 It can be seen from the three groups of curves shown that the curves obtained after compensation by liquid standards and the curves obtained by solid standards are not only different in value, but also, taking the first sample as an example, when the temperature begins to change (before 25°C), the change amplitude of the first solid compensation curve 111 is significantly greater than that of the first liquid compensation curve 112. After the temperature reaches 26°C, the first liquid compensation curve 112 is relatively stable, and the change with temperature fluctuation is not large, but the fluctuation of the first solid compensation curve 111 with temperature change is significantly greater than that of the first liquid compensation curve 112. Therefore, solid standards have obvious advantages over liquid standards in manufacturing, storage, transportation and use, but in actual measurement, solid standards are significantly more affected by temperature changes than liquid standards.

[0050] On the basis of the prior art, this embodiment adds a second solid standard 12 to the detection part 10. The second solid standard 12 is adjacent to the first solid standard 11 to ensure that the temperature environment is close. In addition, the fluorescence properties of the first solid standard 11 and the second solid standard 12 do not show a linear correlation with the temperature change trend. According to the description of the following embodiment, N groups of calibration data groups need to be introduced when fitting the first mapping relationship. If the fluorescence properties of the first solid standard 11 and the second solid standard 12 are linearly correlated, the first fluorescence ratio of each group of calibration data groups is the same, resulting in the first fluorescence ratio being independent of the temperature change of the solid standard and unable to achieve the compensation effect. Therefore, the first solid standard 11 and the second solid standard 12 of this embodiment belong to different substances and meet the following conditions: (1) The fluorescence intensity of the two is not linearly correlated with the temperature change; (2) The first fluorescence ratio is as large as possible to reduce the system error; (3) The relationship between the first fluorescence ratio and its own temperature change can remain stable for a long time to ensure the stability of the system. Those skilled in the art can select the specific substance of the solid standard according to the above conditions and their own needs, and no limitation is made here.

[0051] The following is based on Figure 1 The optical detection device shown is used to further illustrate the technical solution of the embodiment of the present invention.

[0052] Reference Figure 2 , Figure 2 A flowchart of a fluorescence value compensation method for a solid-state standard provided in an embodiment of the present invention includes but is not limited to the following steps:

[0053] S10, at the calibration temperature, perform N scanning operations through the SiPM optical component to obtain N groups of calibration data groups, wherein the calibration data group includes a first calibration fluorescence value and a second calibration fluorescence value obtained based on the same scan, the first calibration fluorescence value corresponds to the first solid standard, the second calibration fluorescence value corresponds to the second solid standard, and N is a positive integer.

[0054] It should be noted that the calibration temperature is different from the reference temperature in the following step S20. The calibration temperature can be any working temperature other than the reference temperature. After starting the optical detection equipment, the detection process is run to change the working temperatures of the first solid-state standard, the second solid-state standard and the SiPM optical component. The temperature at the start of the scan is recorded. If it is different from the preset reference temperature, it can be determined as the calibration temperature. The scanning operation is continued. The first calibration fluorescence value is recorded when scanning the first solid-state standard, and the second calibration fluorescence value is recorded when scanning the second solid-state standard. A set of first calibration fluorescence values ​​and second calibration fluorescence values ​​are obtained each time the scan is performed. A total of N calibration data sets are obtained after N executions.

[0055] It should be noted that, since the temperature of the SiPM components will change during the scanning process, the calibration temperature of N scans may be different, and the relevant temperature can be recorded before each scan.

[0056] It is worth noting that the PCR process usually includes two temperature points: denaturation and annealing and extension. The calibration temperature of this embodiment is obtained after the annealing and extension temperature is completed, and then the relevant scanning operation is performed.

[0057] In addition, in one embodiment, referring to Figure 1 The scanning operation specifically includes but is not limited to the following steps:

[0058] The first light source is turned off, and the first solid-state standard is scanned based on the SiPM optical component to obtain a system dark background value; the first light source is turned on, and the first solid-state standard is scanned based on the SiPM optical component to obtain a first detection fluorescence value, and the second solid-state standard is scanned to obtain a second detection fluorescence value; a first target fluorescence value is obtained based on the first detection fluorescence value and the system dark background value, and a second target fluorescence value is obtained based on the second detection fluorescence value and the system dark background value; wherein, when the scanning operation is performed at the calibration temperature, the first target fluorescence value is a first calibration fluorescence value, and the second target fluorescence value is a second calibration fluorescence value; or, when the scanning operation is performed at the reference temperature, the first target fluorescence value is a first reference fluorescence value, and the second target fluorescence value is a second reference fluorescence value; or, when the scanning operation is performed after starting sample detection, the first target fluorescence value is a first measured fluorescence value, and the second target fluorescence value is a second measured fluorescence value.

[0059] It should be noted that when performing a scanning operation, the influence of background noise needs to be considered, that is, the dark background value of the system in this embodiment. When the light source is turned off, the dark background at each location is the same. Therefore, this embodiment does not limit the scanning point, and the dark background value obtained by scanning the SiPM optical component can be used as the system dark background value. Among them, the fluorescence value satisfies the following formula: F=I×Gain+B, wherein F is the output current measured by the system, that is, the first detection fluorescence value or the second detection fluorescence value of this embodiment, B is the background noise of the system, that is, the system dark background value of this embodiment, Gain is the gain of SiPM at the current operating temperature and voltage, I is the emission light intensity of the solid standard, and the actual fluorescence value of the solid standard needs to propose a dark background value based on the detection fluorescence value, so it can be obtained that f=FB=I×Gain, wherein f is the first target fluorescence value or the second target fluorescence value.

[0060] It should be noted that, in this embodiment, each time a scanning operation is performed at any stage, the first light source is first turned off to scan and obtain the corresponding dark background value, and then the first light source is turned on to scan and obtain the detected fluorescence value. Finally, the target fluorescence value is calculated by the difference between the detected fluorescence value and the dark background value. The same operation is performed on the first solid standard object and the second solid standard object to obtain corresponding parameters, and different parameters are obtained at different stages. For example, the first target fluorescence value obtained in the calibration stage is the first calibration fluorescence value, and the rest will not be elaborated.

[0061] S20, at a preset reference temperature, performing a scanning operation through the SiPM optical component to obtain a first reference fluorescence value and a second reference fluorescence value, wherein the first reference fluorescence value corresponds to a first solid standard, and the second reference fluorescence value corresponds to a second solid standard.

[0062] It should be noted that the reference temperature can be set in advance as the target temperature for compensation of the first solid-state standard. Through the technical solution of this embodiment, the fluorescence value of the first solid-state standard at any temperature can be equivalent to the reference temperature, thereby improving the accuracy of temperature compensation. Based on this, the first solid-state standard and the second solid-state standard can be controlled to be at the reference temperature, and a scan is performed to obtain the first reference fluorescence value and the second reference fluorescence value, which serve as the data basis for subsequent calculations and construction of relationship equations.

[0063] S30, determining the ratio of the first calibration fluorescence value and the second calibration fluorescence value of the same group as the first fluorescence ratio of the calibration data group, and constructing a first mapping relationship based on the first calibration fluorescence value, the first reference fluorescence value and the first fluorescence ratio, wherein the first mapping relationship is used to characterize the mapping ratio of the first calibration fluorescence value and the first reference fluorescence value.

[0064] It should be noted that after measuring N groups of calibration data groups, the ratio of the first calibration fluorescence value to the second calibration fluorescence value of each group of calibration data groups is determined as the first fluorescence ratio. The first fluorescence ratio can characterize the difference in fluorescence values ​​of two different solid-state standards under the same temperature environment. Therefore, in the case of multiple first fluorescence ratios, the difference in fluorescence values ​​of two different solid-state standards under various temperature environments can be reflected. Since the first solid-state standard and the second solid-state standard are not linearly correlated with temperature changes, the difference is determined by the difference in the characteristics of the solid-state standards. By using the first fluorescence ratio to erase the difference, the stable fluorescence value of the first solid-state standard at different temperatures can be obtained. Therefore, this embodiment does not perform temperature compensation on the SiPM, but compensates the fluorescence value of the first solid-state standard by the second solid-state standard, so that the fluorescence value of the first solid-state standard is closer to the liquid standard that is less affected by temperature, thereby improving the accuracy of subsequent temperature compensation.

[0065] It should be noted that, since there are N groups of calibration data groups, when there are enough first fluorescence ratios, the difference in fluorescence values ​​between the first solid standard and the second solid standard under various temperature environments can be characterized by multiple first fluorescence ratios. On this basis, the first reference fluorescence value is combined as a quantification and the first calibration fluorescence value is used as a variable to construct a first mapping relationship, so that when the first calibration fluorescence value has different values, the first calibration fluorescence value can be numerically compensated in combination with the first reference fluorescence value and the first mapping relationship. The obtained first compensated fluorescence value can be mapped to the reference temperature. Although it is not necessarily equal to the numerical value of the first reference fluorescence value, compared with before compensation, the deviation from the compensation effect of the liquid standard has been effectively improved.

[0066] like Figure 4 As shown, the three groups of samples to be tested are the fourth sample, the fifth sample and the sixth sample. Before temperature compensation, the curve of the fourth sample is the fourth pre-compensation curve 123, the curve of the fifth sample is the fifth pre-compensation curve 223, and the curve of the sixth sample is the sixth pre-compensation curve 323. When only the first solid standard is used for compensation, the curve of the fourth sample is the fourth pre-correction curve 121, the curve of the fifth sample is the fifth pre-correction curve 221, and the curve of the sixth sample is the sixth pre-correction curve 321. It can be seen that although the three groups of pre-correction curves tend to be stable, they still have large fluctuations with changes in temperature. After correction by the second solid standard, the curve of the fourth sample is the fourth post-correction curve 122, the curve of the fifth sample is the fifth post-correction curve 222, and the curve of the sixth sample is the sixth post-correction curve 322. Compared with the corresponding pre-correction curves, the corrected curves of each sample are more stable under temperature changes.

[0067] In addition, in one embodiment, constructing a first mapping relationship based on the first calibrated fluorescence value, the first reference fluorescence value, and the first fluorescence ratio specifically includes but is not limited to the following steps:

[0068] A first light intensity ratio is determined based on a first calibrated fluorescence value and a first reference fluorescence value, wherein the first light intensity ratio is used to indicate a ratio of a first calibrated optical intensity to a first reference optical intensity, the first calibrated optical intensity being the emission light intensity of a first solid-state standard at a calibration temperature, and the first reference optical intensity being the emission light intensity of the first solid-state standard at a reference temperature; a first mapping relationship is constructed based on the first light intensity ratio and the first fluorescence ratio.

[0069] It should be noted that the first light intensity ratio can characterize the difference between the emission light intensity of the first solid-state standard at various calibration temperatures and the reference temperature. After the first light source irradiates the first solid-state standard, the first solid-state standard emits light after being excited by the light signal, and its emission light intensity has a first calibration optical intensity or a first reference optical intensity. The first calibration fluorescence value or the first reference fluorescence value is only a numerical value obtained by equivalently outputting the system current, and cannot accurately characterize the fluorescence intensity of the first solid-state standard itself, that is, the first calibration fluorescence value is not equal to the first reference optical intensity. Although the emission light intensity cannot be measured directly, according to the above expression f=I×Gain, I can be characterized by f and Gain. Based on this, using the characteristic of Gain maintaining stability, it can be obtained ,Right now , Subscripts are used to indicate that the parameter belongs to the first solid state standard. is the calibration temperature, Based on this, in this embodiment, the ratio of the first calibrated fluorescence value to the first reference fluorescence value is equal to the ratio of the first calibrated optical intensity to the first reference optical intensity, thereby achieving the equivalent replacement of the emission light intensity of the first solid standard by the measured fluorescence value.

[0070] It should be noted that, according to the description of the above embodiment, the first fluorescence ratio can be used as the data basis for the second solid-state standard to compensate the first solid-state standard. A first mapping relationship is constructed based on the first fluorescence ratio and the first light intensity ratio. After measuring the fluorescence value at any temperature, the specific value of the first light intensity ratio can be calculated according to the first mapping relationship. The first light intensity ratio is the conversion ratio between the fluorescence value and the reference value. Therefore, the first light intensity ratio can be used to convert the fluorescence value into the value at the reference temperature, thereby realizing temperature compensation of the first solid-state standard.

[0071] In addition, in one embodiment, constructing a first mapping relationship according to the first light intensity ratio and the first fluorescence ratio specifically includes but is not limited to the following steps:

[0072] A first mapping relationship is constructed based on the first light intensity ratio and the first fluorescence ratio, wherein the first mapping relationship includes multiple fitting coefficients; based on the first reference fluorescence value, N first calibrated fluorescence values ​​and N first fluorescence ratios, a fitting calculation is performed on the first mapping relationship to determine the values ​​of each fitting coefficient.

[0073] In addition, in one embodiment, the expression of the first mapping relationship is:

[0074] ;in, is the first calibration optical intensity, is the first reference optical intensity, is the calibration temperature, is the reference temperature, For indicating the first solid standard, { , , , ···} is the fitting coefficient, x is the first fluorescence ratio, and satisfies , is the first calibration fluorescence value, is the second calibration fluorescence value, is the first light intensity ratio.

[0075] It should be noted that the properties of solid fluorescent materials are different from those of liquid fluorescent materials. The law of change of the fluorescence value of solid fluorescent materials with temperature can be described by Taylor expansion as follows: , T is the fluorescence value at any temperature, I is the emission light intensity of the corresponding solid standard, at the same time and in the same optical system, the temperature of the two solid standards is approximately the same, and the gain Gain of SiPM is the same as the dark background B. α and β represent the first solid standard and the second solid standard respectively, and according to the above Taylor expansion, we can get: . Accordingly, refer to The principle can be obtained ,in, is the emission light intensity of the first solid standard after being excited by the light signal, is the emission intensity of the second solid standard after being excited by the light signal, is the fluorescence value of the first solid standard after the dark background is removed, that is, the first calibrated fluorescence value, the first reference fluorescence value or the first measured fluorescence value, The same goes for each fluorescence value of the second solid standard.

[0076] On this basis, a first mapping relationship can be constructed based on the first light intensity ratio and the first fluorescence ratio, that is, ; The above formula is the first mapping relationship of this embodiment, where x= Therefore, the relevant data of the second solid standard is introduced into the calculation process, so as to realize the compensation of the first solid standard by the second solid standard.

[0077] It should be noted that, in this embodiment, the first light intensity ratio is used to compensate the first measured fluorescence value of the first solid standard, so the specific value of the first light intensity ratio needs to be calculated. Before that, the specific values ​​of the fitting coefficients in the first mapping relationship need to be determined. , the first calibrated fluorescence value and the second calibrated fluorescence value are known, and there are N sets of data, so the N first fluorescence ratios can be respectively substituted into the first mapping relationship to obtain an equation in which N unknowns are only fitting coefficients, for example Figure 5 As shown in FIG. 1 , after substituting N first fluorescence ratios, the first mapping relationship is fitted into an ascending curve, and then the first mapping relationship is fitted according to the conventional least square method to calculate the values ​​of each fitting coefficient. If the first solid standard is to be used to compensate the second solid standard, the corresponding second mapping relationship is referenced to Figure 5 The curve with a downward trend calculates the fitting coefficients in the same way, which will not be repeated here. In addition, the fitting operation is a technique well known to those skilled in the art, which will not be explained here one by one.

[0078] S40, after starting the sample detection, performing a scanning operation through the SiPM optical component to obtain a first measured fluorescence value of the first solid standard and a second measured fluorescence value of the second solid standard, and determining a first compensated fluorescence value based on the first measured fluorescence value, the second measured fluorescence value and the first mapping relationship, wherein the first compensated fluorescence value is used to characterize the mapping value of the first measured fluorescence value at the reference temperature.

[0079] In addition, in one embodiment, determining the first compensated fluorescence value based on the first measured fluorescence value, the second measured fluorescence value and the first mapping relationship specifically includes but is not limited to the following steps:

[0080] Determine a second fluorescence ratio based on the first measured fluorescence value and the second measured fluorescence value; replace the first fluorescence ratio in the first mapping relationship with the second fluorescence ratio to calculate the value of the first light intensity ratio; determine a first compensation fluorescence value based on the first measured fluorescence value and the first light intensity ratio.

[0081] It should be noted that, when { , , , ···}, the first mapping relationship is ,and Therefore, at any temperature, knowing the fluorescence values ​​of the first solid standard and the second solid standard, we can substitute them as x to calculate The specific value of is the value of the first light intensity ratio. ,by , is the temperature of the sample during testing, is the first measured fluorescence value, is the second measured fluorescence value, y is the second fluorescence ratio, and the second fluorescence ratio is equivalent to the first fluorescence ratio, except that the temperature described in the construction of the relationship is the calibration temperature and the measured temperature, so the first mapping relationship can be replaced by Based on this, at any measured temperature, we only need to obtain the first measured fluorescence value and the second measured fluorescence value in the same scan and then insert them into the above formula to obtain , and then calculate the first compensated fluorescence value equivalent to the reference temperature of the first measured fluorescence value. = / ( ), , is the first compensation fluorescence value.

[0082] It should be noted that after obtaining the first compensated fluorescence value, temperature compensation of the SiPM component is performed according to the fluorescence value of the solid standard at the reference temperature, which is a solution in the prior art and will not be elaborated herein.

[0083] In addition, in one embodiment, after the first reference fluorescence value and the second reference fluorescence value are obtained by performing a scanning operation through the SiPM optical component, the following steps are specifically included but not limited to:

[0084] The ratio of the second calibration fluorescence value and the first calibration fluorescence value of the same group is determined as the second fluorescence ratio of the calibration data group, and a second mapping relationship is constructed based on the second calibration fluorescence value, the second reference fluorescence value and the second fluorescence ratio, wherein the second mapping relationship is used to characterize the mapping ratio of the second calibration fluorescence value and the second reference fluorescence value; after starting the sample detection, the first measured fluorescence value of the first solid standard and the second measured fluorescence value of the second solid standard are obtained through the SiPM optical component, and a second compensation fluorescence value is determined based on the first measured fluorescence value, the second measured fluorescence value and the second mapping relationship, wherein the second compensation fluorescence value is used to characterize the mapping value of the second measured fluorescence value at the reference temperature.

[0085] It should be noted that the first solid-state standard and the second solid-state standard are two adjacent solid-state standards arranged in the detection section, and according to the relevant technology, the detection section includes a plurality of samples to be tested, so for some samples to be tested that are closer to the first solid-state standard, the first solid-state standard can be used for temperature compensation, and for some samples to be tested that are closer to the second solid-state standard, the second solid-state standard can be used for temperature compensation. Therefore, according to the description of the above embodiment, the fluorescence values ​​of the first solid-state standard and the second solid-state standard can be obtained simultaneously during each scan, so according to the same principle, the first solid-state standard can be used to perform temperature compensation on the second solid-state standard, and then the second compensated fluorescence value of the compensated second solid-state standard can be used to perform subsequent SiPM temperature compensation, thereby realizing mutual compensation of multiple solid-state standards.

[0086] It should be noted that the principle of constructing the second mapping relationship for the second solid standard is the same as that of the first mapping relationship. The only difference is that the positions of the relevant numerical values ​​of the first solid standard and the second solid standard are swapped, which will not be repeated here.

[0087] like Figure 6 As shown, Figure 6 : is a structural diagram of a fluorescence value compensation device for a solid-state standard provided by an embodiment of the present invention. The present invention also provides a fluorescence value compensation device for a solid-state standard, comprising:

[0088] The processor 601 may be implemented by a general-purpose central processing unit (CPU), a microprocessor, an application-specific integrated circuit (ASIC), or one or more integrated circuits, and is used to execute relevant programs to implement the technical solutions provided in the embodiments of the present application;

[0089] The memory 602 can be implemented in the form of a read-only memory (ROM), a static storage device, a dynamic storage device or a random access memory (RAM). The memory 602 can store an operating system and other application programs. When the technical solution provided in the embodiment of this specification is implemented by software or firmware, the relevant program code is stored in the memory 602, and the processor 601 calls and executes the fluorescence value compensation method of the solid-state standard in the embodiment of this application;

[0090] Input / output interface 603, used to implement information input and output;

[0091] Communication interface 604, used to realize communication interaction between the device and other devices, which can be realized through wired mode (such as USB, network cable, etc.) or wireless mode (such as mobile network, WIFI, Bluetooth, etc.);

[0092] A bus 606 that transmits information between various components of the device (e.g., the processor 601, the memory 602, the input / output interface 603, and the communication interface 604);

[0093] The processor 601 , the memory 602 , the input / output interface 603 and the communication interface 604 are connected to each other in communication within the device via a bus 605 .

[0094] An embodiment of the present application further provides an electronic device, comprising the fluorescence value compensation device of the solid-state standard as described above.

[0095] The embodiment of the present application further provides a storage medium, which is a computer-readable storage medium and stores a computer program. When the computer program is executed by a processor, the fluorescence value compensation method of the solid-state standard is implemented.

[0096] As a non-transient computer-readable storage medium, the memory can be used to store non-transient software programs and non-transient computer executable programs. In addition, the memory may include a high-speed random access memory, and may also include a non-transient memory, such as at least one disk storage device, a flash memory device, or other non-transient solid-state storage devices. In some embodiments, the memory may optionally include a memory remotely arranged relative to the processor, and these remote memories may be connected to the processor via a network. Examples of the above-mentioned networks include, but are not limited to, the Internet, an intranet, a local area network, a mobile communication network, and a combination thereof. The device embodiments described above are merely schematic, wherein the units described as separate components may or may not be physically separated, and are implemented to be located in one place, or may also be distributed to multiple network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the present embodiment.

[0097] It will be appreciated by those skilled in the art that all or some of the steps and systems in the methods disclosed above may be implemented as software, firmware, hardware, and appropriate combinations thereof. Some or all physical components may be implemented as software executed by a processor, such as a central processing unit, a digital signal processor, or a microprocessor, or as hardware, or as an integrated circuit, such as an application-specific integrated circuit. Such software may be distributed on a computer-readable medium, which may include a computer storage medium (or non-transitory medium) and a communication medium (or transient medium). As known to those skilled in the art, the term computer storage medium includes volatile and non-volatile, removable and non-removable media implemented in any method or technology for storing information (such as computer-readable instructions, data structures, program modules, or other data). Computer storage media include, but are not limited to, RAM, ROM, EEPROM, flash memory or other memory technology, CD-ROM, digital versatile disk (DVD) or other optical disk storage, magnetic cassettes, magnetic tapes, disk storage or other magnetic storage devices, or any other medium that can be used to store desired information and can be accessed by a computer. Furthermore, it is well known to those skilled in the art that communication media typically include computer readable instructions, data structures, program modules, or other data in a modulated data signal such as a carrier wave or other transport mechanism, and may include any information delivery media.

[0098] The above is a specific description of the preferred implementation of the present invention, but the present invention is not limited to the above-mentioned implementation mode. Technical personnel familiar with the field can also make various equivalent deformations or substitutions under the shared conditions without violating the spirit of the present invention. These equivalent deformations or substitutions are all included in the scope defined by the claims of the present invention.

Claims

1. A method for compensating the fluorescence value of a solid standard, characterized in that: The invention is applied to an optical detection device, wherein the optical detection device comprises a SiPM optical component and a detection unit, wherein the detection unit is provided with a first solid-state standard object and a second solid-state standard object adjacent to each other, wherein the fluorescence values ​​of the first solid-state standard object and the second solid-state standard object are not linearly correlated with temperature changes, and the fluorescence value compensation method of the solid-state standard object comprises: At a calibration temperature, performing N scanning operations through the SiPM optical component to obtain N groups of calibration data sets, wherein the calibration data sets include a first calibration fluorescence value and a second calibration fluorescence value obtained based on the same scan, the first calibration fluorescence value corresponds to the first solid-state standard, the second calibration fluorescence value corresponds to the second solid-state standard, and N is a positive integer; At a preset reference temperature, performing a scanning operation through the SiPM optical component to obtain a first reference fluorescence value and a second reference fluorescence value, wherein the first reference fluorescence value corresponds to the first solid-state standard, and the second reference fluorescence value corresponds to the second solid-state standard; Determine the ratio of the first calibration fluorescence value and the second calibration fluorescence value of the same group as the first fluorescence ratio of the calibration data group, and construct a first mapping relationship based on the first calibration fluorescence value, the first reference fluorescence value and the first fluorescence ratio, wherein the first mapping relationship is used to characterize the mapping ratio of the first calibration fluorescence value and the first reference fluorescence value; After starting the sample detection, a scanning operation is performed through the SiPM optical component to obtain a first measured fluorescence value of the first solid standard and a second measured fluorescence value of the second solid standard, and a first compensation fluorescence value is determined based on the first measured fluorescence value, the second measured fluorescence value and the first mapping relationship, wherein the first compensation fluorescence value is used to characterize a mapping value of the first measured fluorescence value at the reference temperature; Constructing a first mapping relationship based on the first calibrated fluorescence value, the first reference fluorescence value, and the first fluorescence ratio includes: Determining a first light intensity ratio based on the first calibration fluorescence value and the first reference fluorescence value, wherein the first light intensity ratio is used to indicate a ratio of a first calibration optical intensity to a first reference optical intensity, the first calibration optical intensity being the emission light intensity of the first solid standard at the calibration temperature, and the first reference optical intensity being the emission light intensity of the first solid standard at the reference temperature; constructing the first mapping relationship according to the first light intensity ratio and the first fluorescence ratio; The expression of the first mapping relationship is: ;in, is the first calibration optical intensity, is the first reference optical intensity, is the calibration temperature, is the reference temperature, for indicating the first solid standard, { , , , ···} is the fitting coefficient, x is the first fluorescence ratio, and satisfies , is the first calibrated fluorescence value, is the second calibrated fluorescence value, is the first light intensity ratio.

2. The fluorescence value compensation method of a solid standard according to claim 1, characterized in that: The SiPM optical component is provided with a first light source, and the scanning operation includes: Turn off the first light source, and obtain a system dark background value based on scanning of the SiPM optical component; Turning on the first light source, scanning the first solid standard object based on the SiPM optical component to obtain a first detection fluorescence value, and scanning the second solid standard object to obtain a second detection fluorescence value; Obtain a first target fluorescence value based on the first detected fluorescence value and the system dark background value, and obtain a second target fluorescence value based on the second detected fluorescence value and the system dark background value; Wherein, when the scanning operation is performed at the calibration temperature, the first target fluorescence value is the first calibration fluorescence value, and the second target fluorescence value is the second calibration fluorescence value; or, when the scanning operation is performed at the reference temperature, the first target fluorescence value is the first reference fluorescence value, and the second target fluorescence value is the second reference fluorescence value; or, when the scanning operation is performed in the actual measurement stage, the first target fluorescence value is the first measured fluorescence value, and the second target fluorescence value is the second measured fluorescence value.

3. The fluorescence value compensation method of a solid standard according to claim 1, characterized in that: Constructing the first mapping relationship according to the first light intensity ratio and the first fluorescence ratio includes: constructing a first mapping relationship based on the first light intensity ratio and the first fluorescence ratio, wherein the first mapping relationship includes a plurality of fitting coefficients; Based on the first reference fluorescence value, the N first calibration fluorescence values ​​and the N first fluorescence ratios, a fitting calculation is performed on the first mapping relationship to determine the value of each of the fitting coefficients.

4. The fluorescence value compensation method of a solid standard according to claim 1, characterized in that: Determining a first compensated fluorescence value based on the first measured fluorescence value, the second measured fluorescence value, and the first mapping relationship includes: determining a second fluorescence ratio based on the first measured fluorescence value and the second measured fluorescence value; replacing the first fluorescence ratio in the first mapping relationship with the second fluorescence ratio, and calculating a value of the first light intensity ratio; The first compensated fluorescence value is determined based on the first measured fluorescence value and the first light intensity ratio.

5. The fluorescence value compensation method of a solid standard according to any one of claims 1 to 4, characterized in that: After performing a scanning operation through the SiPM optical component to obtain a first reference fluorescence value and a second reference fluorescence value, the method further includes: Determine the ratio of the second calibration fluorescence value to the first calibration fluorescence value of the same group as the second fluorescence ratio of the calibration data group, and construct a second mapping relationship based on the second calibration fluorescence value, the second reference fluorescence value and the second fluorescence ratio, wherein the second mapping relationship is used to characterize the mapping ratio of the second calibration fluorescence value to the second reference fluorescence value; After starting sample detection, a first measured fluorescence value of the first solid-state standard and a second measured fluorescence value of the second solid-state standard are obtained through the SiPM optical component, and a second compensated fluorescence value is determined based on the first measured fluorescence value, the second measured fluorescence value and the second mapping relationship, wherein the second compensated fluorescence value is used to characterize the mapping value of the second measured fluorescence value at the reference temperature.

6. A fluorescence value compensation device for a solid standard, characterized in that: It includes at least one control processor and a memory for communicating with the at least one control processor; the memory stores instructions that can be executed by the at least one control processor, and the instructions are executed by the at least one control processor so that the at least one control processor can execute the fluorescence value compensation method of the solid-state standard as described in any one of claims 1 to 5.

7. An electronic device, characterized in that: A fluorescence value compensation device comprising the solid state standard as described in claim 6.

8. A computer-readable storage medium, characterized in that: The computer-readable storage medium stores computer-executable instructions, and the computer-executable instructions are used to enable a computer to execute the fluorescence value compensation method of a solid-state standard according to any one of claims 1 to 5.

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