Self-reference alignment and optical path compensation structure of microfluidic fluorescence immunoassay card and detection method thereof
Patent Information
- Application Number
- CN202611097481.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-07-23
- Publication Date
- 2026-10-09
AI Technical Summary
[0005]本发明的目的在于提供一种微流控荧光免疫检测卡匣的自基准对位与光程补偿结构及其检测方法，以解决卡匣插入偏差、检测窗口偏移、窗口透过率差异和光程变化导致检测信号波动，而现有方案不能在结果换算前进行结构化识别、补偿和无效拦截的问题
[0009]与现有技术相比，本发明的有益效果在于：第一，通过插卡定位边形成卡匣插入定位基础，减少插卡深度和横向偏差；第二，通过自基准对位标记识别检测窗口相对于图像采集视场的位置，使检测前能够发现窗口偏移、旋转或比例异常；第三，通过透过率参考区和光程补偿区补偿窗口污染、透过率差异、厚度差异和光路距离变化造成的信号波动；第四，通过补偿前置门控，避免在卡匣未到位、对位失败、参考异常或补偿超限时输出看似有效的浓度结果。
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Figure CN122882732A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the fields of microfluidic detection and fluorescence immunoassay, and particularly to a self-reference alignment and optical path compensation structure for a microfluidic fluorescence immunoassay cartridge and its detection method. Background Technology
[0002] Microfluidic fluorescence immunoassay cartridges typically need to be inserted into an analyzer. The analyzer's light source and imaging modules excite and acquire data from the cartridge's detection window, and the results processing module calculates the detection results based on the fluorescence signal. For miniaturized, desktop, or near-patient testing devices, the cartridge insertion position, detection window position, window transmittance, and changes in optical path directly affect the stability of the fluorescence signal.
[0003] In existing solutions, some cartridges rely solely on the mechanical fit between the outer casing and the card slot for positioning. Due to factors such as cartridge injection molding dimensional tolerances, window bonding thickness, contamination and scratches during use, improper card insertion by the user, cartridge tilting, or optical path assembly errors, the position of the detection window relative to the light source module and imaging module may shift. If the analyzer directly reads the signal from the main reaction zone and calls the standard curve, it may mistake cartridge position deviations, window transmittance differences, or optical path differences for sample concentration differences.
[0004] Therefore, a structure and method are needed that combines card insertion positioning, detection window self-reference alignment, window transmittance reference, and optical path compensation. This allows the analyzer to confirm whether the card is in place, whether the window is in the allowed position, and whether the transmittance reference and optical path compensation are normal before outputting the detection results. When the conditions are met, the main reaction area signal is compensated, and when there are abnormalities, the invalid results are hard locked. Summary of the Invention
[0005] The purpose of this invention is to provide a self-reference alignment and optical path compensation structure for a microfluidic fluorescence immunoassay cartridge and its detection method, so as to solve the problem that the detection signal fluctuates due to cartridge insertion deviation, detection window offset, window transmittance difference and optical path change, and that existing solutions cannot perform structured identification, compensation and invalid interception before result conversion.
[0006] To achieve the above objectives, this invention provides a self-reference alignment and optical path compensation structure, including a cartridge body and an analyzer-side light source module, imaging module, and result processing module that cooperate with the cartridge body. The cartridge body has a cartridge positioning edge, a sample inlet, a detection window, a flow channel, and a main reaction area. The sample inlet is connected to the main reaction area within the detection window via the flow channel. The detection window, or its adjacent area, also includes a transmittance reference area, an optical path compensation area, and one or more self-reference alignment marks.
[0007] The light source module provides excitation light to the detection window, and the imaging module acquires images within the acquisition field of view during the same detection cycle. The image acquisition field of view covers the main reaction area, the transmittance reference area, the optical path compensation area, and the self-reference alignment mark. The result processing module determines the alignment state of the detection window relative to the image acquisition field of view based on the self-reference alignment mark, judges the transmittance state of the window based on the transmittance reference area, and forms the optical path compensation parameters based on the optical path compensation area.
[0008] Only when the card insertion positioning state, alignment state, transmittance reference state, and optical path compensation parameters all meet the preset conditions, the result processing module uses the optical path compensation parameters to correct the main reaction zone signal and calls the standard curve or threshold rule to output the detection result. If any condition is not met, calling the standard curve or threshold rule is prohibited, and the detection result is locked as invalid.
[0009] Compared with the prior art, the beneficial effects of the present invention are as follows: First, by forming the card insert positioning base through the card insertion positioning edge, the insertion depth and lateral deviation are reduced; Second, by identifying the position of the detection window relative to the image acquisition field of view through the self-reference alignment mark, window offset, rotation or proportional abnormality can be detected before detection; Third, by compensating for signal fluctuations caused by window contamination, transmittance differences, thickness differences and changes in optical path distance through the transmittance reference area and optical path compensation area; Fourth, by pre-compensation gating, seemingly effective concentration results are avoided when the card is not in place, alignment fails, reference is abnormal or compensation exceeds the limit. Attached Figure Description
[0010] Figure 1 This is a schematic diagram of the cartridge body structure.
[0011] Figure 2 This is a structural diagram of an optical detection system.
[0012] Figure 3 This is a magnified view of a portion of the detection window.
[0013] Figure 4 This is a flowchart of the detection method. Detailed Implementation
[0014] The embodiments of the present invention will now be described with reference to the accompanying drawings. These embodiments are used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention.
[0015] like Figure 1As shown, this embodiment provides a self-reference alignment and optical path compensation structure for a microfluidic fluorescence immunoassay cartridge. The cartridge body 1 has a card insertion positioning edge 2, a sample inlet 3, a detection window 4, a main reaction area 5, a transmittance reference area 6, an optical path compensation area 7, a self-reference alignment mark 8, and a flow channel 9. The sample inlet 3 is connected to the main reaction area 5 within the detection window 4 via the flow channel 9, allowing the sample to enter the corresponding reaction area of the detection window 4 along the flow channel 9. The card insertion positioning edge 2 is used to cooperate with the analyzer's card slot or positioning component, so that the cartridge body 1 forms a repeatable detection position after insertion.
[0016] The main reaction region 5 is used to generate a fluorescence detection signal related to the target analyte. The transmittance reference region 6 is located within or near the detection window 4 and does not participate in the target analyte-specific immunoreaction; it provides a window transmittance reference signal. The optical path compensation region 7 is located within or near the detection window 4 and provides a compensation reference signal related to changes in window thickness, assembly gap, focal length deviation, or optical path distance. The self-reference alignment mark 8 provides a reference for identifying the position of the detection window 4 within the image acquisition field of view.
[0017] like Figure 2 As shown, the light source module 10 and the imaging module 11 are respectively paired with the detection window 4. The light source module 10 is used to provide excitation light to the detection window 4, and the imaging module 11 is used to acquire images of the area where the detection window 4 is located. The image acquisition field of view 13 covers the main reaction area 5, the transmittance reference area 6, the optical path compensation area 7, and the self-reference alignment mark 8, so that alignment recognition, transmittance reference acquisition, optical path compensation calculation, and main reaction area result correction can be completed within the same detection cycle. The result processing module 12 receives or processes the image information acquired by the imaging module 11, and completes alignment judgment, reference judgment, compensation calculation, and result release based on the image information.
[0018] In one embodiment, the self-reference alignment mark 8 can be a cross mark, a ring mark, a corner mark, a bar mark, a hole mark, or a combination thereof. The result processing module 12 can calculate the translation, rotation angle, tilt state, or proportional deviation of the detection window 4 in the image acquisition field of view 13 based on the geometric relationship between the self-reference alignment mark 8 and the boundary of the detection window 4. When the self-reference alignment mark 8 is missing, recognition fails, the positional deviation exceeds the limit, or the boundary of the detection window 4 is incomplete, it is determined to be an alignment anomaly.
[0019] like Figure 3As shown, the local area of detection window 4 includes a main reaction area 5, a transmittance reference area 6, an optical path compensation area 7, a self-reference alignment mark 8, and a flow channel 9. After the sample enters the corresponding area of detection window 4 along the flow channel 9, the main reaction area 5 generates a detection signal; the transmittance reference area 6 provides a reference signal to determine whether there is contamination, obstruction, scratches, or abnormal transmittance in detection window 4; and the optical path compensation area 7 provides a compensation reference signal to generate optical path compensation parameters. Figure 3 The two self-reference alignment marks 8 are different settings of the same type of component, which are used to help identify the posture of the detection window 4.
[0020] Each batch of cartridges can have its standard position, attitude tolerance, allowable range of transmittance reference area 6, and parameter range of optical path compensation area 7 determined before leaving the factory, and these parameters are bound to the cartridge batch information. The batch information of the current cartridge can be read through the QR code, barcode, or other encoding on the cartridge body 1, or it can be entered through the analyzer interface. The result processing module 12 calls up the corresponding parameters according to the current batch information.
[0021] The result processing module 12 can calculate the corrected main reaction zone signal based on the original signal of the main reaction zone 5, the background signal of the main reaction zone, the signal of the transmittance reference zone 6, and the signal of the optical path compensation zone 7. The corrected main reaction zone signal is used to enter the standard curve, segmented threshold rule, or result mapping rule of the corresponding detection item. If the transmittance reference zone 6 signal is below the lower limit, saturated, or fluctuates beyond the limit, or if the compensation parameter formed by the optical path compensation zone 7 is not within the allowable range of the current cartridge batch, the standard curve or threshold rule is prohibited from being called.
[0022] like Figure 4 As shown, the detection method of this embodiment includes the following steps: S1, card insertion and positioning: insert the card cartridge body 1 into the analyzer and complete the positioning through the card insertion and positioning edge 2; S2, alignment recognition: identify the self-reference alignment mark 8 within the image acquisition field of view 13 and determine the alignment status of the detection window 4; S3, single window acquisition: perform image acquisition on the same detection window 4 by the light source module 10 and the imaging module 11; S4, transmittance reference acquisition: acquire the transmittance reference area 6 signal and determine whether it is within the allowable range of the corresponding card cartridge batch; S5, optical path compensation calculation: acquire the optical path compensation area 7 signal and calculate the optical path compensation parameters; S6, main reaction area result correction: when all conditions are met, correct the main reaction area 5 signal using the optical path compensation parameters; S7, result release: input the corrected main reaction area signal into the standard curve or threshold rule of the corresponding item and output the detection result.
[0023] When alignment anomalies, reference anomalies, or compensation exceeding limits occur, the result processing module 12 locks the current test result as invalid and prompts for retesting. The above structure and method bind card insertion positioning, window self-reference identification, transmittance reference, optical path compensation, and result release within the same testing cycle, avoiding the problem of relying solely on subsequent software smoothing, empirical correction, or weighted processing without the support of a card holder structure and optical reference. Without departing from the technical concept of this invention, any equivalent substitutions or modifications made to the above structural form, mark shape, compensation area material, optical path form, and parameter calling method should be included within the scope of protection of this invention.
Claims
1. A self-reference alignment and optical path compensation structure for a microfluidic fluorescence immunoassay cartridge, comprising a cartridge body and an analyzer-side light source module, an imaging module, and a result processing module that cooperate with the cartridge body, wherein the cartridge body is provided with a cartridge positioning edge, a sample inlet, a detection window, a flow channel, and a main reaction area, characterized in that, The sample inlet is connected to the main reaction area within the detection window via the flow channel. The detection window or its adjacent area is also provided with a transmittance reference area, an optical path compensation area, and one or more self-reference alignment marks. The self-reference alignment marks and the boundary of the detection window together form an alignment reference for the detection window within the image acquisition field of view. The light source module is configured to provide excitation light to the detection window. The imaging module is configured to acquire images within the image acquisition field of view within the same detection cycle. The image acquisition field of view covers the main reaction area, the transmittance reference area, the optical path compensation area, and the self-reference alignment mark. The result processing module is configured to determine at least one alignment state of the detection window relative to the image acquisition field of view, including positional offset, rotational deviation, or proportional deviation, based on the self-reference alignment mark; determine the window transmittance state based on the transmittance reference area; form optical path compensation parameters based on the optical path compensation area; and only when the card insertion positioning state, alignment state, transmittance reference state, and optical path compensation parameters all meet preset conditions, correct the main reaction area signal using the optical path compensation parameters and output the detection result. If any condition is not met, the standard curve or threshold rule shall not be invoked, and the result of this test shall be locked as invalid.
2. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, The card insertion positioning edge is located at the insertion end or side area of the card cartridge body and cooperates with the positioning surface or elastic pre-tightening member in the card insertion slot of the analyzer, so that the card cartridge body maintains at least one of the following positioning states during the detection cycle: insertion depth, lateral position, and detection window height.
3. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, The self-reference alignment mark is a cross mark, a ring mark, a corner mark, a bar mark, a hole mark, or a combination thereof. The result processing module determines the translation amount, rotation angle, tilt state, or proportional deviation of the detection window based on the geometric relationship between the self-reference alignment mark and the boundary of the detection window.
4. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, The transmittance reference area is a transparent area, a semi-transparent area, a fixed reference film area, or a microporous reference area that does not participate in the immune reaction of the target analyte and has a preset optical transmittance. The result processing module determines whether the detection window is contaminated, obstructed, scratched, or has abnormal transmittance based on the acquisition signal of the transmittance reference area.
5. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, The optical path compensation region includes a preset optical path length region, a stepped light transmission region, a fixed scattering reference region, a stripe reference region, or a combination thereof, and is used to provide a compensation reference signal related to changes in the thickness of the detection window, assembly gap, focal length deviation, or optical path distance.
6. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, The main reaction area, transmittance reference area, and optical path compensation area are arranged sequentially along the sample flow direction or the length of the detection window, and are all located within the same image acquisition field of view of the imaging module, so that the transmittance reference signal, optical path compensation signal, and main reaction area signal are acquired within the same detection cycle.
7. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, The result processing module stores the standard position of the self-reference alignment mark, attitude tolerance, transmittance reference range and optical path compensation parameter range for the corresponding cartridge batch, and calls the corresponding parameters according to the current cartridge batch information.
8. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, The result processing module calculates the corrected main reaction zone signal based on the original signal of the main reaction zone, the background signal of the main reaction zone, the transmittance reference zone signal, and the optical path compensation zone signal. The corrected main reaction zone signal is used to enter the standard curve, segmented threshold rule, or result mapping rule of the corresponding detection item.
9. The self-reference alignment and optical path compensation structure according to claim 1, characterized in that, Before outputting the detection results, the result processing module sequentially performs card insertion positioning judgment, alignment recognition judgment, single window acquisition integrity judgment, transmittance reference anomaly judgment, and optical path compensation parameter over-limit judgment. If any judgment fails, the standard curve or threshold rule call interface is kept in a prohibited state, and the detection result is locked as invalid.
10. A method for detection using the self-reference alignment and optical path compensation structure according to any one of claims 1 to 9, characterized in that, include: S1, Insert the card cartridge body into the analyzer and complete the card insertion positioning through the card insertion positioning edge; S2, Identify the self-reference alignment mark within the image acquisition field of view and determine the alignment status of the detection window; S3, The light source module and imaging module perform single-window acquisition on the same detection window to obtain an image containing the main reaction area, transmittance reference area, optical path compensation area, and self-reference alignment mark; S4, Acquire the transmittance reference area signal and determine whether the transmittance reference area signal is within the allowable range of the corresponding cartridge batch; S5, Acquire the optical path compensation area signal and calculate the optical path compensation parameter, and determine whether the optical path compensation parameter is within the preset range; S6, When the conditions for card insertion positioning, alignment identification, transmittance reference, and optical path compensation are all met, the optical path compensation parameter is used to correct the result of the main reaction area signal; S7, input the corrected main reaction zone signal into the standard curve or threshold rule of the corresponding item and output the detection result; When alignment anomalies, reference anomalies, or compensation exceeding limits occur, the standard curve or threshold rule should not be invoked, the current test result should be locked as invalid, and a retest should be prompted. The alignment anomalies include card insertion positioning not meeting preset conditions or failure to recognize the self-reference alignment mark, and the reference anomalies include incomplete image acquisition field of view or abnormal transmittance reference area signal.