Bronchoalveolar lavage fluid rapid detection method based on fluorescent staining technology

Abstract

The invention relates to a bronchoalveolar lavage fluid rapid detection method based on a fluorescent staining technology and a corresponding detection device, and aims to improve the detection efficiency of alveolar lavage and sampling screening. The detection device comprises an endoscope assembly, a lavage fluid supply module, a recovery module and a control module. Wherein the recovery module is internally provided with an assembly used for adding a fluorescent coloring agent into the recovery liquid, the fluorescent coloring agent is injected into the recovery liquid in the recovery process of the lavage fluid according to needs, and an electromagnetic drive stirring mechanism is combined, so that dynamic and uniform mixing of the dyeing liquid and the lavage fluid is achieved. The control module automatically adjusts the injection amount, the mixing speed and the stirring duration of the coloring agent according to preset parameters or real-time detection data, and the dyeing reaction time in the operation process is remarkably saved.

Description

Technical Field

[0001] The present invention belongs to the field of medical detection technology, and in particular relates to a rapid detection method for bronchoalveolar lavage fluid based on fluorescence staining technology. Background Art

[0002] In the clinical examination of lung diseases, bronchoalveolar lavage (BAL) is a commonly used sampling method that can directly obtain alveolar fluid for cytological analysis, pathogen detection, and determination of immune markers. However, the traditional BAL fluid testing process has problems such as complicated operating steps, time-consuming manual staining, and insufficient mixing of dye solutions, resulting in a long testing cycle and difficulty meeting the clinical needs for rapid diagnosis. Especially in scenarios such as acute respiratory infections, immune pneumonia, or large-scale epidemic screening, methods that rely on laboratory staining and processing can no longer meet the requirements of rapid response. In addition, most existing devices are separate structures, and lavage, staining, and testing require the transfer of samples between multiple devices, increasing the risk of contamination and error. Therefore, there is an urgent need for a BAL detection system that is more integrated, more automated, and suitable for rapid fluorescent staining and processing to improve overall detection efficiency and standardization.

[0003] After consulting relevant public technologies, the technical solution with publication number CN113274563B proposes an irrigation catheter and a corresponding irrigation catheter system to improve the convenience of suction during irrigation operations; the technical solution with publication number TW201637672A proposes a diversion-type irrigation head structure, which improves the sealing of the irrigation head at the connection through an improved mechanical structure; the technical solution with publication number US12251530B2 proposes an automatic bronchial irrigation and suction device for clearing sputum or other secretions in the patient's bronchi.

[0004] The above technical solutions all propose several technical solutions to improve the convenience and work efficiency of irrigation operations, but currently there are few technical solutions that specifically discuss the relevant inspection processes using fluorescence detection technology.

[0005] The foregoing discussion of the background art is intended only to facilitate an understanding of the present invention. This discussion does not acknowledge or admit that any of the material referred to is part of the common general knowledge. Summary of the Invention

[0006] The purpose of the present invention is to provide a rapid detection method for bronchoalveolar lavage fluid based on fluorescent staining technology and a corresponding detection device, aiming to improve the detection efficiency of alveolar lavage and sampling screening. The detection device includes an endoscope component, an lavage fluid supply module, a recovery module and a control module. Among them, the recovery module is provided with a component for adding fluorescent dye to the recovered fluid, and the fluorescent dye is injected into the recovered fluid during the lavage fluid recovery process as needed, and combined with an electromagnetically driven stirring mechanism, dynamic and uniform mixing of the dye and the lavage fluid is achieved. The control module automatically adjusts the injection amount of the dye, the mixing speed and the stirring duration according to preset parameters or real-time detection data, significantly saving the dyeing reaction time in the operation process.

[0007] The present invention adopts the following technical solution: a rapid detection method for bronchoalveolar lavage fluid based on fluorescence staining technology, the detection method comprising the following steps: S100: estimating the target analyte to be detected based on the basic pathology or pre-screening test results of the subject, selecting one or more fluorescent dyes based on the target analyte, and disposing each fluorescent dye into an independent liquid storage chamber; S200: Inputting a plurality of preset control parameters into a control module; S300: Perform bronchoalveolar lavage on the patient and recover the lavage fluid; S400: As the perfusion fluid is recovered and injected into the recovery tank, the control module injects a specified fluorescent dye into the recovery tank according to preset parameters, and continuously stirs and mixes the mixture of the recovery fluid and the fluorescent dye; S500: When the recovery liquid reaches the preset volume, sufficient mixing time is maintained and the mixed recovery liquid is transferred to the next stage of detection.

[0008] Preferably, during the execution of step S400, the detection method further includes performing the following operations: The mixed liquid in the recovery tank is quickly tested to update the control parameters in real time and execute the instructions of the updated control parameters.

[0009] Preferably, the preset parameters include at least: Injection parameters, including the injection dose and injection timing of the fluorescent dye; The stirring parameters include at least one of the stirring mode, stirring speed, and stirring duration.

[0010] Preferably, the mixed liquid in the recovery tank is rapidly tested, and the rapid test includes one or more of the following parameters: The current weight of the recovered liquid; Fluorescence reaction of the current recovery solution.

[0011] At the same time, a rapid detection device for bronchoalveolar lavage fluid based on fluorescent staining technology is proposed, and the detection device is applied to the detection method to implement the detection operation; the detection device includes: An endoscope assembly, comprising an operating handle and a guide tube extending therefrom, the guide tube being provided with a working channel for injection and recovery of lavage fluid, and through which an endoscope imaging assembly can be introduced to acquire images of the interior of the bronchus; an irrigating fluid supply module, comprising an infusion set connected to the infusion port of the operating handle, for injecting irrigating fluid into the patient's bronchus; The recovery module includes a negative pressure source for generating negative pressure, a recovery liquid container assembly, and a negative pressure circuit connected to the guide tube; the recovery liquid container assembly includes a recovery tank, and the negative pressure source generates negative pressure in the recovery tank, thereby sucking the irrigation liquid into the recovery tank, thereby realizing the recovery of the irrigation liquid; The recovery liquid container assembly further includes a fluorescent dye injection subassembly; the fluorescent dye injection subassembly includes at least one liquid storage cavity for temporarily storing the fluorescent dye and an extruder, and the extruder is used to squeeze the liquid storage cavity, so that the dye stored in the liquid storage cavity enters the recovery tank; The recovery liquid container assembly also includes a stirring and mixing subassembly; the stirring and mixing subassembly includes an electromagnetic driver disposed below the recovery tank and a magnetic mixing block at the bottom of the recovery cavity inside the recovery tank; the electromagnetic driver drives the mixing block in a non-contact manner to mix the recovery liquid and the dye after the recovery liquid is injected into the recovery tank; In addition, the detection device further includes the control module, which is used to receive and store multiple control parameters. The control module is electrically connected to the extruder and the electromagnetic driver, and generates control instructions to control the extruder and the electromagnetic driver to work.

[0012] The beneficial effects achieved by the present invention are: This technical solution integrates a fluorescent dye injection mechanism and an electromagnetically driven stirring mechanism within the recovery fluid container, enabling simultaneous dye injection and liquid mixing during the lavage fluid recovery process. Compared to the traditional multi-step process of separation and dyeing, transfer, and waiting for mixing, this significantly shortens dye preparation time and improves emergency detection capabilities.

[0013] This technical solution dynamically monitors the weight of the recovered liquid in the recovery tank to automatically adjust the dye extrusion amount and mixing speed. This feedback control mechanism effectively optimizes the dye addition ratio, avoiding over- or under-dosing, while maintaining good mixing and improving dyeing uniformity.

[0014] The technical solution allows the configuration of multiple dye storage chambers, supports rapid fluorescent labeling detection of different target pathogens, and is suitable for rapid screening of Mycoplasma pneumoniae, respiratory syncytial virus, influenza virus, etc. It has good versatility and clinical practicality, and will help promote the practical application of bedside rapid detection equipment.

[0015] The software and hardware parts of the detection device of this technical solution adopt a modular design. The various working modules and components of the hardware part of the system, as well as the instructions, parameters, and algorithms of the software part can be easily replaced and / or upgraded at a later stage, thereby reducing the construction cost and maintenance cost of the system. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The present invention can be further understood from the following description in conjunction with the accompanying drawings. The components in the figures are not necessarily drawn to scale, but rather the emphasis is placed on illustrating the principles of the embodiments. In different views, the same reference numerals designate corresponding parts.

[0017] Explanation of the accompanying reference numerals: 100 - detection device; 101 - endoscope assembly; 102 - recovery liquid container assembly; 104 - negative pressure device; 106 - processing device; 110 - operating handle; 111 - guide tube; 112 - infusion pump; 121 - input terminal; 122 - negative pressure terminal; 123 - infusion port; 204 - recovery tank; 210 - functional base; 211 - electromagnetic drive; 212 - mixing block; 222 - recovery chamber; 224 - liquid storage chamber; 226 - squeezer; 228 - injection port; 230 - one-way permeable membrane; 302 - pressure-sensitive sensor; 500 - computer system; 502 - bus; 504 - processor; 506 - main memory; 508 - read-only memory; 510 - storage device; 512 - display; 514 - input device; 516 - cursor control device; 518 - network device; Figure 1 This is a flow chart of the steps of the detection method of the present invention; Figure 2 Schematic diagram of the framework of the detection device of the present invention; Figure 3 Schematic diagram of the appearance of the recovery liquid container assembly according to an embodiment of the present invention; Figure 4 Schematic diagram of the internal structure of the recovery liquid container assembly according to an embodiment of the present invention; Figure 5 Schematic diagram of the framework of the computer system used by the processing device in the embodiment of the present invention. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with its embodiments; it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. For those skilled in the art, other systems, methods and / or features of the present embodiment will become apparent after reviewing the following detailed description. It is intended that all such additional systems, methods, features and advantages are included in this specification. Included within the scope of the present invention and protected by the appended claims. Additional features of the disclosed embodiments are described in the following detailed description, and these features will be apparent from the following detailed description.

[0019] The same or similar reference numerals in the drawings of the embodiments of the present invention correspond to the same or similar components. In the description of the present invention, it should be understood that if there are terms such as "upper", "lower", "left", "right", etc. indicating an orientation or positional relationship, they are based on the orientation or positional relationship shown in the drawings. This is only for the convenience of describing the present invention and simplifying the description, and does not indicate or imply that the device or component referred to must have a specific orientation. The terms used in the drawings to describe the positional relationship are only for illustrative purposes and cannot be understood as limiting this patent. For those skilled in the art, the specific meanings of the above terms can be understood according to the specific circumstances.

[0020] Example 1: For example, a rapid detection method for bronchoalveolar lavage fluid based on fluorescence staining technology is proposed, as shown in the attached Figure 1 As shown, the detection method includes the following steps: S100: estimating the target analyte to be detected based on the basic pathology or pre-screening test results of the subject, selecting one or more fluorescent dyes based on the target analyte, and disposing each fluorescent dye into an independent liquid storage chamber; S200: Inputting a plurality of preset control parameters into a control module; S300: Perform bronchoalveolar lavage on the patient and recover the lavage fluid; S400: As the perfusion fluid is recovered and injected into the recovery tank, the control module injects a specified fluorescent dye into the recovery tank according to preset parameters, and continuously stirs and mixes the mixture of the recovery fluid and the fluorescent dye; S500: When the recovery liquid reaches the preset volume, sufficient mixing time is maintained and the mixed recovery liquid is transferred to the next stage of detection.

[0021] Preferably, during the execution of step S400, the detection method further includes performing the following operations: The mixed liquid in the recovery tank is quickly tested to update the control parameters in real time and execute the instructions of the updated control parameters.

[0022] Preferably, the preset parameters include at least: Injection parameters, including the injection dose and injection timing of the fluorescent dye; The stirring parameters include at least one of the stirring mode, stirring speed, and stirring duration.

[0023] Preferably, the mixed liquid in the recovery tank is rapidly tested, and the rapid test includes one or more of the following parameters: The current weight of the recovered liquid; Fluorescence reaction of the current recovery solution.

[0024] At the same time, a rapid detection device for bronchoalveolar lavage fluid based on fluorescent staining technology is proposed, and the detection device is applied to the detection method to implement the detection operation; the detection device includes: An endoscope assembly, comprising an operating handle and a guide tube extending therefrom, the guide tube being provided with a working channel for injection and recovery of lavage fluid, and through which an endoscope imaging assembly can be introduced to acquire images of the interior of the bronchus; an irrigating fluid supply module, comprising an infusion set connected to the infusion port of the operating handle, for injecting irrigating fluid into the patient's bronchus; The recovery module includes a negative pressure source for generating negative pressure, a recovery liquid container assembly, and a negative pressure circuit connected to the guide tube; the recovery liquid container assembly includes a recovery tank, and the negative pressure source generates negative pressure in the recovery tank, thereby sucking the irrigation liquid into the recovery tank, thereby realizing the recovery of the irrigation liquid; The recovery liquid container assembly further includes a fluorescent dye injection subassembly; the fluorescent dye injection subassembly includes at least one liquid storage cavity for temporarily storing the fluorescent dye and an extruder, and the extruder is used to squeeze the liquid storage cavity, so that the dye stored in the liquid storage cavity enters the recovery tank; The recovery liquid container assembly also includes a stirring and mixing subassembly; the stirring and mixing subassembly includes an electromagnetic driver disposed below the recovery tank and a magnetic mixing block at the bottom of the recovery cavity inside the recovery tank; the electromagnetic driver drives the mixing block in a non-contact manner to mix the recovery liquid and the dye after the recovery liquid is injected into the recovery tank; In addition, the detection device further includes the control module, which is used to receive and store multiple control parameters. The control module is electrically connected to the extruder and the electromagnetic driver, and generates control instructions to control the extruder and the electromagnetic driver to work.

[0025] For example, as shown in the attached Figure 1FIG2 is a schematic diagram of a detection device 100 using the detection method of the present invention. The detection device 100 includes an endoscope assembly 101 and a recovery fluid container assembly 102 for containing alveolar lavage recovery fluid.

[0026] The endoscope assembly 101 includes an operating handle 110 and a guide tube 111 extending from the detection end of the operating handle 110. The operating handle 110 includes a gripping end and a detection end. When used for detection, the operating handle 110 is held and operated by medical staff to control the extension distance, pointing rotation angle and other postures of the guide tube 111. The guide tube 111 includes a working channel extending from the internal part of the operating handle 110 to the distal end of the guide tube 111. The working channel includes a first channel for accommodating the pipeline portion of the endoscope, and a second channel for conveying irrigation fluid. For example, the attached Figure 1 Only a portion of the guide tube 111 is shown. The recovery fluid container assembly 102 is connected to the manifold inside the operating handle 110 via a conduit via an input port 121 and further communicates with the second channel of the guide tube 111. Meanwhile, the negative pressure port 122 of the recovery fluid container assembly 102 is connected to the negative pressure device 104 via a conduit.

[0027] The operating handle 110 also includes an infusion port 123, which is connected to the guide tube 111 via a manifold provided within the operating handle 110. The user operates the infusion pump 112 to pressure-inject saline solution for alveolar lavage into the infusion port 123, which then flows through the guide tube 111 into the patient's bronchi.

[0028] At this time, the negative pressure device 104 is activated to introduce negative pressure into the recovery fluid container assembly 102 through the negative pressure terminal 122, thereby creating a negative pressure environment inside the recovery fluid container assembly 102. At this time, the saline solution used for irrigating the patient's body, along with the body fluids that have been flushed out of the bronchus, enters the guide tube 111 and is drawn into the recovery fluid container assembly 102.

[0029] Preferably, the endoscope's tubing can be introduced into the patient's bronchus via a guide tube 111 to capture images of the patient's bronchus, and image acquisition can be performed during the bronchoalveolar lavage procedure. Preferably, the endoscope assembly 101 includes necessary imaging components, such as an image acquisition module, an illumination source, and image signal transmission circuitry. Preferably, the image acquisition module can include a CMOS or CCD image sensor, located at or near the distal end of the guide tube 111, for capturing real-time images of the patient's bronchial lining. The illumination source can utilize a fiber optic light guide or a micro-LED array to provide sufficient illumination within the bronchial lumen. The captured image signals can be transmitted via the signal transmission circuitry embedded within the guide tube 111 to the operating handle 110. The operating handle 110 is then connected to the processing device 106 via an external image transmission cable 110. The image information within the bronchial lumen is displayed in real time on the display screen of the processing device 106, which also stores the image information and allows for further analysis.

[0030] Through the imaging element, the user can observe the front end position of the guide tube 111 and the anatomical structure characteristics of the bronchus in real time during the lavage operation, thereby achieving visual guidance and effectively improving the safety and accuracy of the alveolar lavage operation. Preferably, the detection device 100 may further include one or more flow meters configured to measure the amount of fluid output from the infusion pump 112 and / or the amount of fluid recovered from the distal end of the working channel.

[0031] When actually operating the detection device 100 to perform bronchial lavage on a patient, the user can control the distal end of the guide tube 111 to extend into the internal position of the patient's bronchus by operating the operating handle 110, including controlling the depth of the distal end of the guide tube 111 by pushing and pulling back the guide tube 111, and controlling the forward position of the distal end of the guide tube 111 by flipping the operating handle 110.

[0032] At the same time, the user can obtain real-time images of the patient's bronchi through the endoscope to observe the surface features of the patient's bronchi and use the obtained images to assist in the lavage operation.

[0033] Attachment Figure 2 The connecting cables, catheters, etc. shown in the figure are schematic settings. Their actual length needs to be set to a sufficient length based on the actual situation of the detection device 100, and should be set so as not to affect the user's irrigation operation and endoscope operation.

[0034] Further, as attached Figure 3 The figure shows the appearance of the recovery liquid container assembly 102. Figure 4FIG. 1 is a cross-sectional schematic diagram of the recovery liquid container assembly 102 , used to schematically illustrate its internal functional structure.

[0035] Exemplarily, the functional base 210 is located below the recovery tank 204, supporting and covering the bottom of the recovery tank 204. Preferably, the functional base 210 is made of polycarbonate (PC) or ABS engineering plastic to provide space for the various components within. Preferably, the recovery tank 204 can be removed from the functional base 210 and replaced, making it disposable.

[0036] Exemplarily, an electromagnetic driver 211 is provided inside the functional base 210. The electromagnetic driver 211 is configured to generate an alternating magnetic field for driving the mixing block 212 to rotate at the bottom of the recovery chamber 222 in a non-contact manner, thereby stirring the liquid in the recovery chamber 222 and achieving liquid mixing. Preferably, the electromagnetic driver can be composed of a Hall control coil, a multi-pole magnetic core structure, or a rotating magnetic ring array, so that the electromagnetic driver 211 can generate a magnetic field change of the required frequency and amplitude under the control signal instruction, driving the mixing block 212 to produce stable rotation behavior.

[0037] The mixing block 212 is located at the bottom of the recovery chamber 222 and is made of a magnetic material. The bottom of the mixing block is smooth and has a density roughly equal to that of water, allowing the mixing block 212 to be semi-suspended in the liquid. The mixing block 212 rotates under the electromagnetic drive of the electromagnetic driver 211, stirring the recovered liquid and promoting rapid mixing of the recovered liquid with other solutions.

[0038] Exemplarily, the functional base 210 is internally provided with a power module (not shown). In some embodiments, the power module is powered by a built-in rechargeable battery. Preferably, the built-in battery can be a lithium polymer battery pack and provided with a charging port for charging from an external charging device. In other embodiments, the built-in battery can be provided with a contactless magnetic charging module, which charges the built-in battery via an external power supply coil.

[0039] In other embodiments, the power module is connected to an external power supply device via a power interface, converts the power into direct current, and supplies power to the various electrical components in the functional base 210. When an external power supply is used, the power input port on the outer wall of the functional base 210 can be connected to an AC power conversion module to convert the external power into a low-voltage DC power supply suitable for the operation of the electromagnetic actuator.

[0040] Exemplarily, to adjust the liquid stirring operation within the recovery chamber, a control unit is further provided in the functional base 210. The control unit is electrically coupled to the electromagnetic driver 211 and is provided with a data communication port or a wireless receiving unit capable of responding to control commands sent by an external control device (e.g., a control display device or an independent control panel). Preferably, the control unit can adjust the following parameters of the electromagnetic driver 211 in real time: Magnetic field frequency, magnetic field strength, start and stop time; and based on the above basic parameters, program control can be performed, such as setting various types of action modes, including constant speed rotation, intermittent stirring or step-by-step acceleration control logic.

[0041] Exemplarily, a liquid storage chamber 224 is provided on the outer wall of the recovery chamber 222. Liquid storage chamber 224 is a flexible structure, preferably made of a silicone capsule or other medical material with suitable elasticity, and is used to store a predetermined volume of dye solution. A liquid injection port 228 is provided at the top of liquid storage chamber 224. This injection port 228 can utilize an elastic sealing structure, allowing an operator to inject the dye solution into liquid storage chamber 224 using a syringe while preventing the ingress of external air.

[0042] For example, a one-way permeable membrane 230 is partially disposed on the contact surface between the liquid storage chamber 224 and the recovery chamber 222. The one-way permeable membrane 230 is a microporous structure with one-way permeability. It only allows the dye liquid to pass through the membrane under the action of the squeezer 226 and enter the recovery chamber 222, while preventing the recovered liquid from flowing back into the liquid storage chamber 224. In addition, the liquid storage chamber 224 can be exposed to the outside when the recovery tank 204 is not installed on the functional base 210, so that the user can easily replace it and achieve a disposable use.

[0043] Preferably, the number of the liquid storage chambers 224 can be 1, 2, 4 or more. In use, the liquid storage chamber 224 can be installed as needed, for example, when a coloring agent needs to be added, only one liquid storage chamber 224 can be installed.

[0044] Preferably, an extruder 226 is provided on one side of the functional base 210 near the liquid storage chamber 224. The extruder 226 is connected to the control unit. The extruder 226 is a controllable drive component that, upon receiving a control signal, applies pressure to the liquid storage chamber 224, forcing the dye solution through the unidirectional permeable membrane and into the recovery chamber 222. By setting the operating parameters of the extruder 226, the dosage, timing, and sequence of dye solution injection can be precisely controlled.

[0045] Preferably, the squeezer 226 is a micro-drive structure to achieve a controllable squeezing force on the liquid storage chamber 224 in a small space. In some embodiments, the squeezer is in the form of a piezoelectric actuator or an electric micro-cam mechanism to achieve squeezing of the liquid storage chamber 224.

[0046] Embodiment 2: This embodiment should be understood to include at least all the features of any one of the aforementioned embodiments and to be further improved thereon.

[0047] The following is an exemplary method of selecting a dye for several cases, which can be applied to the rapid detection device of the present technical solution.

[0048] For example, for patients with bronchial asthma or chronic obstructive pulmonary disease, samples are obtained through bronchoalveolar lavage, and then fluorescently stained for white blood cells using a FITC-labeled anti-CD45 antibody. Under a fluorescence microscope, morphological characteristics (such as granule distribution and nuclear morphology) of inflammatory cells such as eosinophils (coarse orange-red granules in the cytoplasm and minimally lobed nuclei) and neutrophils (small lavender granules in the cytoplasm and hyperlobed nuclei) can be observed. Morphological differences can be quickly classified and statistically analyzed to aid in the determination of the type of airway inflammation.

[0049] For example, for mild fungal lung infections, Calcofluor White fluorescent staining (which specifically binds to the chitin component of the fungal cell wall) is used in combination with SYTO 9 nucleic acid dye to label fungal nucleic acids. Typical morphological features of Aspergillus can be observed under a fluorescence microscope: septate hyphae (distinct transverse septa visible within the hyphae), branching angles of approximately 45°, and apical swellings and radially arranged conidia (round or oval with uniform fluorescence distribution). In Candida infections, pseudohyphae (slender, unseptate hypha-like structures) and blastospores (round or oval, connected to the mother cell by a narrow neck) are visible. Morphological features such as hyphal morphology (septate / absent, branching angle) and spore characteristics (morphology and arrangement) after fluorescent staining can assist in the rapid identification of fungal infection type and pathogen species.

[0050] For example, for pulmonary tuberculosis infection, bronchoalveolar lavage fluid samples are stained with auramine O (or auramine-rhodamine) fluorescence (a dye that specifically binds to mycolic acid components of the M. tuberculosis cell wall). Observation under a fluorescence microscope (blue light excitation) reveals typical morphological characteristics of M. tuberculosis: cells are slender, slightly curved rods (approximately 2-4 μm in length), with some visible branches or beaded structures (due to uneven distribution of intracellular lipid particles). The fluorescent signal is bright yellow-green, scattered or clustered (indicating bacterial density). Observation of bacterial morphology (curvature, branching), arrangement (single / clustered), and fluorescence intensity (strongly positive / weakly positive) combined with morphological characteristics can assist in rapidly determining the presence of M. tuberculosis and infection activity, providing a basis for clinical diagnosis.

[0051] For example, dual fluorescent staining with DAPI (to label cell nuclei) and WGA-FITC (to label cell membrane glycosylation) can be used to assess bronchial epithelial cell integrity, such as for pre- and post-operative monitoring. DAPI clearly reveals cell nuclear morphology (complete round, condensed, or fragmented), while WGA-FITC reveals the continuity (e.g., continuous lines or broken dots) and thickness variations of cell membrane glycosylation. By observing cell morphology (e.g., nuclear-cytoplasmic ratio, membrane integrity), the physiological state of epithelial cells (e.g., degree of damage or repair) can be rapidly assessed.

[0052] In some embodiments, to further optimize the mixing effect of the recovery solution and the dye, the total volume of the recovery solution is measured in real time to implement more precise control.

[0053] Exemplarily, a pressure-sensitive sensor 302 is provided at the bottom of the electromagnetic actuator 211. This sensor 302 generates corresponding changes in resistance based on changes in the overall weight of the recovery tank above it. By dynamically sensing weight data, the total amount of recovered fluid can be indirectly estimated, thereby inferring the volume of recovered irrigating fluid. This real-time weight is used to monitor the progress of fluid collection and provide feedback on whether the recovered fluid collection has met the testing objectives, thereby instructing the operator to continue the irrigating / recovery operation or to cease the relevant operation.

[0054] Furthermore, in some embodiments, the operating parameters of the squeezer and the output power of the electromagnetic drive can be controlled according to the real-time weight of the recovery liquid.

[0055] For example, assuming the current recovery liquid weight is q and the preset target weight is Q, the target speed percentage S of the electromagnetic drive is calculated according to the following formula: r : ; The maximum value of the target speed of the electromagnetic drive is determined according to the specific design of the electromagnetic drive and the mixing block. Therefore, by controlling the target speed percentage S r The speed deviation generated in the actual process can be eliminated by controlling the relative value of the speed; wherein β is the amplification factor, and β can be set so that the maximum speed can be reached when q and Q reach a certain ratio, such as 1 / 3 or 1 / 4.

[0056] For example, the magnetic field strength of the electromagnetic driver increases with the amount of recovered liquid to cope with the possible increase in the internal resistance of the liquid. Preferably, the magnetic field strength B currently applied by the electromagnetic driver is set by the following calculation formula: ; In the above formula, B0 is the basic strength of the magnetic field applied by the electromagnetic driver, and a suitable value can be measured experimentally; γ is the maximum magnetic field strength increment, which can be determined based on the upper limit of the magnetic field generated by the electromagnetic driver and the power supply capacity of the power module; n is the adjustment coefficient, preferably, 1≤n<1.5.

[0057] Exemplarily, the extrusion stroke increase rate L of the extruder is adjusted to maintain uniform addition of the dye. Preferably, the extrusion stroke increase rate L is calculated as follows: ; In the above formula, L0 is the base extrusion rate, which is set by relevant technical personnel; dq / dt is the rate of change of the weight of the recovered liquid per unit time; μ and κ are adjustment amplitude coefficients, which can be set according to the characteristics of the dye and the permeability of the unidirectional permeable membrane. e is the base of the natural logarithm.

[0058] Embodiment 3: This embodiment should be understood to include at least all the features of any one of the aforementioned embodiments and to be further improved thereon.

[0059] Further, as attached Figure 5 , which illustrates an implementation of a computer system 500 used by the processing device 106; the computer system 500 can be applied to the data storage, calculation and result output process of each working module in the identification and judgment system.

[0060] Illustratively, computer system 500 includes a bus 502 or other communication mechanism for communicating information, and one or more processors 504 coupled with bus 502 for processing information; processor 504 may be, for example, one or more general-purpose microprocessors.

[0061] The computer system 500 also includes a main memory 506, such as a random access memory (RAM), a cache, and / or other dynamic storage device, coupled to the bus 502 for storing information and instructions to be executed by the processor 504; the main memory 506 may also be used to store temporary variables or other intermediate information during execution of the instructions to be executed by the processor 504; these instructions, when stored in a storage medium accessible to the processor 504, present the computer system 500 as a special-purpose machine customized to perform the operations specified in the instructions.

[0062] The computer system 500 may also include a read-only memory (ROM) 508 or other static storage device coupled to the bus 502 for storing static information and instructions for the processor 504; a storage device 510 such as a magnetic disk, optical disk, or USB drive (flash drive) will be coupled to the bus 502 for storing information and instructions.

[0063] Furthermore, coupled to the bus 502 may also include a display 512 for displaying various information, data, media, etc., and an input device 514 for allowing a user of the computer system 500 to control, manipulate, and / or interact with the computer system 500 .

[0064] A preferred way to interact with the management system may be through a cursor control device 516, such as a computer mouse or similar control / navigation mechanism.

[0065] Furthermore, the computer system 500 may further include a network device 518 coupled to the bus 502; wherein the network device 518 may include, for example, a wired network card, a wireless network card, a switching chip, a router, a switch, and other components; In general, the terms "engine," "component," "system," "database," etc., used herein may refer to logic embodied in hardware or firmware, or to a collection of software instructions, possibly with entry and exit points, written in a programming language such as Java, C, or C++; software components may be compiled and linked into executable programs, installed in a dynamic link library, or may be written in an interpreted programming language (such as BASIC, Perl, or Python); it will be understood that software components may be callable from other components or from themselves, and / or may be called in response to detected events or interrupts.

[0066] Software components configured to execute on a computing device may be provided on a computer-readable medium, such as a compact disc, digital video disc, flash drive, magnetic disk, or any other tangible medium, or as a digital download (and may be initially stored) in a compressed or installable format that requires installation, decompression, or decryption prior to execution); such software code may be stored in part or in whole on a memory device of the executing computing device for execution by the computing device; software instructions may be embedded in firmware, such as an EPROM; it will also be understood that hardware components may be composed of connected logic units (such as gates and flip-flops), and / or may be composed of programmable units (such as a programmable gate array or processor).

[0067] Computer system 500 may implement the techniques described herein using custom hard-wired logic, one or more ASICs or FPGAs, firmware, and / or program logic that, in combination with the computer system, renders computer system 500 a special-purpose computing device.

[0068] In accordance with one or more embodiments, the techniques herein are performed by computer system 500 in response to processor 504 executing one or more sequences of one or more instructions contained in main memory 506; such instructions may be read into main memory 506 from another storage medium, such as storage device 510; execution of the sequences of instructions contained in main memory 506 causes processor 504 to perform the process steps described herein; in alternative embodiments, hard-wired circuitry may be used in place of or in combination with software instructions.

[0069] As used herein, the term "non-transitory media" and similar terms refer to any medium that stores data and / or instructions that cause a machine to operate in a specific manner; such non-transitory media may include non-volatile media and / or volatile media; non-volatile media include, for example, optical or magnetic disks, such as storage device 510; and volatile media include dynamic memory, such as main memory 506.

[0070] Among them, common forms of non-transitory media include, for example, floppy disks, diskettes, hard disks, solid-state drives, magnetic tape or any other magnetic data storage medium, CD-ROMs, any other optical data storage medium, any physical medium with a pattern of holes, RAM, PROM and EPROM, FLASH-EPROM, NVRAM, any other memory chip or cartridge, and networked versions thereof.

[0071] Non-transient media are distinct from, but may be used in conjunction with, transmission media; transmission media participate in the transmission of information between non-transient media; for example, transmission media include coaxial cables, copper wires, and optical fibers, including the wires that comprise bus 502; transmission media may also take the form of sound or light waves, such as radio waves and infrared data communications.

[0072] Although the present application has been described above with reference to various embodiments, it will be understood that many changes and modifications may be made without departing from the scope of the present application. That is, the methods, systems, and devices discussed above are examples. Various configurations may omit, replace, or add various processes or components as appropriate. For example, in alternative configurations, the methods may be performed in an order different from that described, and / or various components may be added, omitted, and / or combined. Moreover, features described with respect to certain configurations may be combined in various other configurations, such as different aspects and elements of the configurations may be combined in a similar manner. Furthermore, as technology develops, the elements therein may be updated, i.e., many of the elements are examples and do not limit the scope of the present disclosure or the claims.

[0073] Specific details are given in the description to provide a thorough understanding of the exemplary configurations, including implementations. However, the configurations can be practiced without these specific details. For example, well-known circuits, processes, algorithms, structures, and techniques have been shown without unnecessary detail to avoid obscuring the configurations. This description provides only example configurations and does not limit the scope, applicability, or configurations of the claims. Instead, the foregoing description of the configurations will provide those skilled in the art with an enabling description for implementing the described techniques. Various changes may be made to the function and arrangement of the elements without departing from the spirit or scope of the present disclosure.

[0074] In summary, it is intended that the above detailed description be considered illustrative rather than restrictive, and it should be understood that the above embodiments are intended to be merely illustrative of the present invention and not to limit the scope of protection of the present invention. After reading the contents of the present invention, a skilled person may make various changes or modifications to the present invention, and these equivalent changes and modifications also fall within the scope defined by the claims of the present invention.

Claims

1. A rapid detection method for bronchoalveolar lavage fluid based on fluorescence staining technology, characterized in that: The detection method comprises the following steps: S100: estimating the target analyte to be detected based on the basic pathology or pre-screening test results of the subject, selecting one or more fluorescent dyes based on the target analyte, and disposing each fluorescent dye into an independent liquid storage chamber; S200: Inputting a plurality of preset control parameters into a control module; S300: Perform bronchoalveolar lavage on the patient and recover the lavage fluid; S400: As the perfusion fluid is recovered and injected into the recovery tank, the control module injects a specified fluorescent dye into the recovery tank according to preset parameters, and continuously stirs and mixes the mixture of the recovery fluid and the fluorescent dye; S500: When the recovery liquid reaches the preset volume, sufficient mixing time is maintained and the mixed recovery liquid is transferred to the next stage of detection.

2. The detection method according to claim 1, wherein During the execution of step S400, the detection method further includes performing the following operations: The mixed liquid in the recovery tank is quickly tested to update the control parameters in real time and execute the instructions of the updated control parameters.

3. The detection method according to claim 1, wherein The preset parameters include at least: Injection parameters, including the injection dose and injection timing of the fluorescent dye; The stirring parameters include at least one of the stirring mode, stirring speed, and stirring duration.

4. The detection method according to claim 1, wherein The mixed liquid in the recovery tank is quickly tested, and the rapid test includes one or more of the following parameters: The current weight of the recovered liquid; Fluorescence reaction of the current recovery solution.

5. A rapid detection device for bronchoalveolar lavage fluid based on fluorescent staining technology, characterized in that: The detection device is applied to the detection method according to any one of claims 1 to 4 to implement the detection operation; the detection device comprises: An endoscope assembly, comprising an operating handle and a guide tube extending therefrom, the guide tube being provided with a working channel for injection and recovery of lavage fluid, and through which an endoscope imaging assembly can be introduced to acquire images of the interior of the bronchus; an irrigating fluid supply module, comprising an infusion set connected to the infusion port of the operating handle, for injecting irrigating fluid into the patient's bronchus; The recovery module includes a negative pressure source for generating negative pressure, a recovery liquid container assembly, and a negative pressure circuit connected to the guide tube; the recovery liquid container assembly includes a recovery tank, and the negative pressure source generates negative pressure in the recovery tank, thereby sucking the irrigation liquid into the recovery tank, thereby realizing the recovery of the irrigation liquid; The recovery liquid container assembly further includes a fluorescent dye injection subassembly; the fluorescent dye injection subassembly includes at least one liquid storage cavity for temporarily storing the fluorescent dye and an extruder, and the extruder is used to squeeze the liquid storage cavity, so that the dye stored in the liquid storage cavity enters the recovery tank; The recovery liquid container assembly also includes a stirring and mixing subassembly; the stirring and mixing subassembly includes an electromagnetic driver disposed below the recovery tank and a magnetic mixing block at the bottom of the recovery cavity inside the recovery tank; the electromagnetic driver drives the mixing block in a non-contact manner to mix the recovery liquid and the dye after the recovery liquid is injected into the recovery tank; In addition, the detection device further includes the control module, which is used to receive and store multiple control parameters. The control module is electrically connected to the extruder and the electromagnetic driver, and generates control instructions to control the extruder and the electromagnetic driver to work.

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