CN111537709A - Shooting detection method and detection device based on time-resolved fluorescence tomography - Google Patents
Shooting detection method and detection device based on time-resolved fluorescence tomography Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5302—Apparatus specially adapted for immunological test procedures
- G01N33/5304—Reaction vessels, e.g. agglutination plates
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
Abstract
The invention discloses a shooting detection method and a detection device based on time-resolved fluorescence tomography, wherein the detection method comprises the following steps: synchronously triggering a timing starting device of the excitation light emitting device and the image acquisition device, so that the timing starting device starts timing while the excitation light emitting device emits excitation light to the detected reagent strip; until the preset time length is reached, the image acquisition device acquires images of the detected reagent strip after the optical filter, light in a specific wavelength range in light reflected by the detected reagent strip passes through the optical filter and enters a lens of the image acquisition device, and light outside the specific wavelength range is filtered by the optical filter and cannot enter the lens of the image acquisition device. In the detection device, the exciting light generated by the specific exciting light emitting device directly acts on the reaction film layer of the reagent strip without special light path design and related equipment; meanwhile, based on the time-resolved fluorescence technology, a dark box and other devices which avoid the interference of ambient light are not needed, and the device can be used under natural light.
Description
Technical Field
The invention relates to the field of biological detection, in particular to a shooting detection method and a detection device based on time-resolved fluorescence chromatography.
Background
Point of care testing (POCT), a new field of medical testing, has gone through nearly 30 years since the 90 s of the last century in china. In recent years, POCT diagnostic products in China are endlessly developed like bamboo shoots in the spring after rain, and a plurality of new products are still put on the market every year. In POCT immunoassay products, detection methods are based on antigen-antibody reactions based on immunization, and in order to realize such immunoassays, antibodies, reaction carriers, labels, and instruments for reaction control and signal reading, etc. are required.
Since the 80 s of the last century, the immunochromatography method which is widely applied to semi-quantitative detection by using colloidal gold labeling is gradually developed into a second generation immunochromatography method which is labeled by using a fluorescent material substance; in order to solve the external interference, improve the detection sensitivity and realize the time-resolved fluorescence immunochromatography with higher detection sensitivity.
In order to achieve high sensitivity and quantitative detection, the time-resolved fluorescence immunochromatographic technology developed on the second generation fluorescence immunochromatographic technology needs special time-resolved fluorescence detection equipment for reading, and the second generation immunochromatographic method using a fluorescent material as a marker can realize quantitative detection. However, the blood sample itself generates fluorescence, which is interfered by scattering of incident light and background fluorescence, so the sensitivity and quantitative analysis ability of the method are limited by noise.
In order to improve the defects of the traditional fluorescent materials, the new generation fluorescent materials with strong background interference resistance are widely applied, such as time-resolved fluorescence chromatography immunoassay as a third generation fluorescence detection method. The technology is characterized in that light waves with one specific wavelength lambda 1 are emitted as an excitation source, and detection is carried out at the wavelength of the other specific wavelength lambda 2, so that the method can effectively remove the influence of the excitation light and has high detection sensitivity. When the technology is applied to antigen detection, the test result is often read by professional reading equipment, the current time-resolved fluorescence chromatography device is completed by adopting a special time-resolved detection instrument, and the instrument internally comprises a lambda 1 wavelength exciting light device capable of emitting specific power, a reagent strip action mechanism, an irradiation reaction bin, a reflected light acquisition device, a central processing unit, a relevant display printing and other peripherals. The existing reading equipment has large volume, complex operation and high cost. When a large amount of samples are screened, the efficiency is low, and the method is not very suitable for screening community users, entry and exit and densely populated places.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a shooting detection method and a detection device based on time-resolved fluorescence chromatography, which can reduce the use threshold, improve the detection speed, reduce the batch-to-batch difference between devices and improve the consistency of repeated detection results, and the technical scheme is as follows:
in one aspect, the invention provides a shooting detection method based on time-resolved fluorescence tomography, which comprises the following steps:
synchronously triggering a timing starting device of an excitation light emitting device and an image acquisition device, so that the timing starting device starts timing while the excitation light emitting device emits excitation light to a detected reagent strip;
until the preset time length is reached, the image acquisition device acquires images of the detected reagent strip after the optical filter, light in a specific wavelength range in light reflected by the detected reagent strip passes through the optical filter and then enters a lens of the image acquisition device, and light outside the specific wavelength range is filtered by the optical filter and cannot enter the lens of the image acquisition device.
Further, the specific wavelength range is set according to a wavelength range of light emitted by excitation of a luminescent substance in a sample on the test reagent strip.
Further, the preset timing duration is set according to a time gap from the excitation of the luminescent substance in the detected sample on the detected reagent strip to the emission of light and/or a decay time of the emitted light.
An optional technical solution is as follows: the timing starting device of the excitation light emitting device and the image acquisition device realizes synchronous triggering by the following method:
starting a flash lamp of the image acquisition device;
when the flash information is detected, triggering the excitation light emitting device to emit excitation light to the detected reagent strip, and simultaneously triggering the timing starting device of the image acquisition device to start timing.
The second optional technical solution is as follows: the timing starting device of the excitation light emitting device and the image acquisition device realizes synchronous triggering by the following method:
and the same trigger signal generator is connected with the exciting light emitting device and the timing starting device in a wired mode, under the triggering action of the trigger signal generator, the exciting light emitting device emits exciting light, and meanwhile, the timing starting device starts timing.
On the other hand, the invention provides a shooting detection device based on time-resolved fluorescence tomography, which comprises an excitation light emitting device for emitting excitation light, an optical filter for filtering interference light, an image acquisition device for acquiring image information of a detected reagent strip, and a synchronous trigger device for synchronizing the excitation light emitting device with a timing starting device of the image acquisition device, wherein the timing starting device is used for starting the image acquisition device to acquire an image of the detected reagent strip at regular time, and the optical filter is arranged between the detected reagent strip and the image acquisition device so that light in a specific wavelength range in the light reflected by the detected reagent strip enters a lens of the image acquisition device after passing through the optical filter;
under the triggering action of the synchronous triggering device, the excitation light emitting device emits excitation light to the detected reagent strip, and the timing starting device starts to time;
and when the preset timing duration is reached, the image acquisition device acquires images of the detected reagent strip.
Further, the synchronous trigger device comprises a controller, a flash lamp arranged at the image acquisition device and a flash receiving device configured to detect flash information, wherein the flash receiving device, the excitation light emitting device and the timing starting device are all electrically connected with the controller;
when the flash receiving device detects flash information, the controller controls to start the excitation light emitting device to emit excitation light and simultaneously controls the timing starting device to start timing.
In still another aspect, the present invention provides a time-resolved fluorescence immunochromatographic system, comprising a reagent strip and the above-mentioned photographing detection apparatus based on time-resolved fluorescence chromatography, wherein the reagent strip is provided with a sample inlet, a reference line region and at least one immunochromatographic region, a sample solution added through the sample inlet is labeled with a specified luminescent material, and a specific wavelength range filtered by an optical filter of the photographing detection apparatus based on time-resolved fluorescence chromatography matches with a wavelength range of light emitted by the specified luminescent material when excited.
Further, the preset time duration of the timing starting device of the photographing detection device based on time-resolved fluorescence tomography is matched with the time gap from the excitation to the light emission of the specified luminescent material and/or the decay time of the emitted light.
In another aspect, the present invention provides an immunochromatography detection method based on the above time-resolved fluorescence immunochromatography system, comprising the following steps:
synchronously triggering a timing starting device of an excitation light emitting device and an image acquisition device, so that the timing starting device starts timing while the excitation light emitting device emits excitation light to a detected reagent strip;
until the preset time length is reached, the image acquisition device acquires images of the detected reagent strip after the optical filter, light in a specific wavelength range in light reflected by the detected reagent strip passes through the optical filter and then enters a lens of the image acquisition device, and light outside the specific wavelength range is filtered by the optical filter and cannot enter the lens of the image acquisition device;
judging whether a reference line area on the reagent strip is displayed or not, if so, continuing to execute the next step, and otherwise, judging that the detection result is invalid;
and obtaining an immunochromatography detection result according to the appearance condition of the corresponding immunochromatography area.
The technical scheme provided by the invention has the following beneficial effects:
a. the detection method of a brand-new photographing mode is used, expensive functional parts are omitted, the operation is simple, the manufacturing cost is low, and the use threshold is reduced;
b. the detection method that a conventional time-resolved detection instrument adopts a scanning reagent strip is changed, and a brand-new detection method of a photographing mode is used, so that the detection speed is improved;
c. the detection result is accurate, the repeatability of the detection result is good, the inter-batch difference between devices is reduced, and the consistency of repeated detection results is improved;
d. no special light path design and related equipment are needed; the device which avoids the interference of the ambient light such as a dark box and the like is not needed, and the device can be directly used under the natural light.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a block diagram of a photographing detection apparatus based on time-resolved fluorescence tomography according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of a photographing detection apparatus based on time-resolved fluorescence tomography according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of one embodiment of the sync trigger of FIG. 2;
FIG. 4 is a schematic diagram showing a first visualization result of the reagent strip obtained by the immunochromatographic assay provided in the embodiment of the present invention;
FIG. 5 is a schematic diagram showing a second visualization result of the reagent strip obtained by the immunochromatographic assay provided in the embodiment of the present invention;
FIG. 6 is a schematic diagram showing a third result of the reagent strip obtained by the immunochromatographic assay provided in the embodiment of the present invention;
FIG. 7 is a schematic diagram showing a fourth visualization result of the reagent strip obtained by the immunochromatographic assay provided in the embodiment of the present invention;
fig. 8 is a schematic structural diagram of a synchronization triggering device in the shooting detection device according to the embodiment of the present invention.
Wherein the reference numerals include: 1-excitation light emission device, 2-optical filter, 3-image acquisition device, 31-timing starting device, 4-reagent strip, 41-sampling hole, 42-datum line region, 43-immunochromatography region and 5-flash receiving device.
Detailed Description
In order to make the technical solutions of the present invention better understood and more clearly understood by those skilled in the art, the technical solutions of the embodiments of the present invention will be described below in detail and completely with reference to the accompanying drawings. It should be noted that the implementations not shown or described in the drawings are in a form known to those of ordinary skill in the art. Additionally, while exemplifications of parameters including particular values may be provided herein, it is to be understood that the parameters need not be exactly equal to the respective values, but may be approximated to the respective values within acceptable error margins or design constraints. It is to be understood that the described embodiments are merely exemplary of a portion of the invention and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. In addition, the terms "comprises" and "comprising," and any variations thereof, in the description and claims of this invention, are intended to cover a non-exclusive inclusion, such that a process, method, apparatus, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The invention aims to provide a time-resolved fluorescence chromatography detection method and a time-resolved fluorescence chromatography detection device based on a mobile terminal photographing method, which meet the requirements of small size, simple operation and easy carrying required by communities and a large number of screening places and realize real-time quantitative detection.
In one embodiment of the invention, a shooting detection device based on time-resolved fluorescence tomography is provided, as shown in FIG. 1, the shooting detection device comprises an excitation light emitting device 1 for emitting excitation light, a filter 2 for filtering interference light, an image acquisition device 3 for acquiring image information of a detected reagent strip, a synchronous trigger device for synchronizing the excitation light emitting device 1 with a timing starting device 31 of the image acquisition device 3, wherein the timing starting device 31 is used for starting the image acquisition device 3 to acquire images of the detected reagent strip 4 at regular time, the optical filter 2 is arranged between the detected reagent strip 4 and the image acquisition device 3, so that light in a specific wavelength range in the light reflected by the detected reagent strip 4 passes through the optical filter 2 and enters the lens of the image acquisition device 3;
under the triggering action of the synchronous triggering device, the excitation light emitting device 1 emits excitation light to the detected reagent strip 4, and the timing starting device 31 starts timing;
and when the preset timing duration is reached, the image acquisition device 3 acquires an image of the detected reagent strip 4. The image acquisition device 3 may be a mobile terminal device, including but not limited to various smart phones, tablet computers, mobile terminal devices with a camera function, and the like, or an image sensor, such as a CCD, a CMOS, and the like.
The method for carrying out time-resolved fluorescence tomography detection on the detected reagent strip 4 by utilizing the shooting detection device based on time-resolved fluorescence tomography comprises the following steps:
synchronously triggering a timing starting device of an excitation light emitting device and an image acquisition device, so that the timing starting device starts timing while the excitation light emitting device emits excitation light to a detected reagent strip;
until the preset time length is reached, the image acquisition device acquires images of the detected reagent strip after the optical filter, light in a specific wavelength range in light reflected by the detected reagent strip passes through the optical filter and then enters a lens of the image acquisition device, and light outside the specific wavelength range is filtered by the optical filter and cannot enter the lens of the image acquisition device.
In the embodiment of the invention, lanthanide series inert elements are taken as an example, and are filled into nano-scale polystyrene microspheres to prepare the microspheres, and the microspheres can be used for antibody detection after being crosslinked with antibodies. The fluorescent microspheres have larger Stock displacement than fluorescein, can effectively eliminate the interference of excitation light and non-specific fluorescence on signal acquisition, improve the specificity of detection, are not easy to be photobleached compared with fluorescein, and have more stable results. Specifically, the specific wavelength range is set according to the wavelength range of the light emitted by the luminescent material in the sample to be tested on the test strip being excited, the preset timing duration is set according to the time gap between the luminescent material in the sample to be tested on the test strip being excited and emitting the light and/or the decay time of the emitted light, in one embodiment, if the excited wavelength of the luminescent material is λ 1, the wavelength of the light emitted after being excited is λ 2, and the time from being excited to emitting the light of λ 2 is τ, the passing wavelength of the optical filter is λ 2, an allowable error range λ 2 ± 10nm which vertically floats around λ 2 may be adopted, and preferably, the allowable error range is λ 2 ± 0.1 nm.
In addition, the fluorescent microspheres have very long fluorescence decay time (500-2000 uS), and the fluorescence signal decay time of fluorescein is very short (5-100 uS), so that the fluorescent microspheres can develop a time resolution technology. The measurement time can be properly delayed, and the measurement can be carried out after the background fluorescence is completely attenuated, so that the interference of exciting light can be eliminated, and the inspection accuracy is improved. The preset timing duration may be τ. It should be noted that, because of the existence of the decay time, the preset timing duration is set to be τ plus a part of the decay time, and as long as the decay time reaches 0%, theoretically, the technical solution of the present invention can be implemented, so that the present invention does not limit the preset timing duration to be exactly τ, but only in the preferred duration, the decay is not started or the decay part is weak, so that the imaging on the image acquisition device is relatively clear, and the detection accuracy is improved.
In one embodiment of the present invention, as shown in fig. 2, the timing starting device 31 of the excitation light emitting device 1 and the image acquisition device 3 realizes synchronous triggering by the following methods:
the same trigger signal generator (i.e. the synchronous trigger device in fig. 2) is connected with the excitation light emitting device 1 and the timing start device 31 by wires, under the trigger action of the trigger signal generator, the excitation light emitting device 1 emits excitation light, and the timing start device 31 starts timing.
In a further embodiment of the invention, as shown in fig. 3, the timing start means 31 of the excitation light emission means 1 and the image acquisition means 3 achieve synchronous triggering by the following means/methods:
the synchronous trigger device comprises a controller, a flash lamp arranged at the image acquisition device 3 and a flash receiving device 5 configured to detect flash information, wherein the flash receiving device 5, the excitation light emitting device 1 and the timing starting device 31 are all electrically connected with the controller;
when the flash light receiving device 5 detects flash light information, the controller controls to start the excitation light emitting device 1 to emit excitation light, and at the same time controls the timing start device 31 to start timing.
The method comprises the following specific steps: starting a flash lamp of the image acquisition device;
when the flash information is detected, the excitation light emitting device is triggered to emit excitation light to the detected reagent strip (the light emitting angle of the excitation light emitting device is adjusted in advance to enable the excitation light emitting device to face the detected reagent strip 4), and the timing starting device 31 of the image acquisition device 3 is triggered to start timing. Until the timing is finished, that is, when the preset timing duration is reached, the timing starting device 31 is used for starting a shutter of the image acquisition device, so that the image acquisition device acquires the image information of the detected reagent strip 4 immediately when the timing duration is reached. Taking the luminescent material as an example, the excitation light emitting device 1 and the timing starting device 31 are triggered synchronously, after the timing duration τ (or a part of the fluorescence decay time may be increased appropriately) is reached, the image collecting device 3 is started, the light wave emitted after the luminescent material is excited reflects from the detected reagent strip through the optical filter 2, since the pre-selected optical filter 2 only allows the light wave with the wavelength λ 2 (which may also float up and down by an error range, such as λ 2 ± 50nm) to pass through, and finally enters the lens of the image collecting device 3, the part to be finally imaged is only the part reflecting the light wave with the wavelength λ 2 (or within the floating error range) on the reagent strip.
In an embodiment of the present invention, referring to fig. 8, the optical filter 2, the flash light receiving device 5, the power supply system, and the annular excitation light emitting device 1 capable of emitting light waves with a wavelength of λ 1 (capable of exciting light-emitting substances) may be integrated, and disposed in front of the lens of the image capturing device 3. Taking a mobile terminal with a shooting function as an example, preferably, the mobile terminal is internally provided with application software, the application software built in the mobile terminal is operated, a key is clicked to detect, according to the requirement of the time-resolved immunochromatography detection method, a flash lamp of the mobile terminal is started according to a parameter set in advance, the flash light receiving device 5 can detect a flash lamp signal of the mobile terminal in real time, and then the excitation light emitting device 1 is triggered to emit excitation light with a certain power of lambda 1 wavelength.
The excitation light having the wavelength λ 1 is irradiated onto the reaction film layer of the test reagent strip 4 according to the optical paths shown in fig. 2 and 3, induces the fluorescent substance on the reaction film layer to emit light, emits emission light having the wavelength λ 2, and is reflected from the reaction film layer as reflected light. The reflected light lambda 2 passes through the optical filter 2 shown in fig. 2 and fig. 3, and interference light with other useless wavelengths is filtered, and then the reflected light is irradiated into a photographing lens of the mobile terminal, and the operation software performs imaging photographing at a specific tau moment according to the requirement of the detection method of the patent to obtain a time-resolved fluorescence image of the reagent strip to be detected. The application software built in the mobile terminal analyzes the obtained time-resolved fluorescence image, compares the reference fluorescence intensity, and gives a final detection result, which is described in detail in the following embodiments.
It should be noted that the excitation light λ 1 depends on which wavelength range of the luminescent material is excited by the light, the reflected light λ 2 depends on which wavelength of the light emitted by the luminescent material after being induced, the specific time τ depends on the time required for the luminescent material to emit the reflected light λ 2 after being excited, and the luminescent material is not limited to the lanthanide provided in the embodiments of the present invention.
The time-resolved fluorescence immunochromatographic detection method can be realized by matching with the above method, and in one embodiment of the present invention, a corresponding time-resolved fluorescence immunochromatographic system is provided, as shown in fig. 2 and 3, the immunochromatographic system comprises a reagent strip 4 and the above photographing detection device based on time-resolved fluorescence chromatography, wherein a sample inlet 41, a reference line region 42 and at least one immunochromatographic region 43 are arranged on the reagent strip 4, a sample solution added through the sample inlet 41 is marked with a specified luminescent substance, and a specific wavelength range filtered by the filter 2 of the photographing detection device based on time-resolved fluorescence chromatography matches with a wavelength range of light emitted by the specified luminescent substance being excited.
As described above, the preset timing duration of the timing start means 31 of the time-resolved fluorescence tomography-based imaging detection apparatus matches the time gap from excitation to emission of the specified luminescent substance and/or the decay time of the emitted light.
Taking the reagent strips shown in FIGS. 4-7 as a distance, the reagent strip 4 shown in the figure has a reference mark line C, an IgM antibody mark line and an IgG antibody mark line. The area of the reagent strip in the oblong shape in the figure, which is located to the left of the reference mark line C, is coated with a luminescent substance; and the same luminescent substance is added to the sample solution to crosslink with the antibody in the form of the above-mentioned microspheres. When the sample solution is added into the sample inlet 41, the sample solution is distributed on the whole reagent strip 4, the region corresponding to the left side of the IgM antibody mark line and the region corresponding to the left side of the IgG antibody mark line on the reagent strip 4 are the immunochromatographic region 43, except that the immunochromatographic region 43 on the left side of the IgM antibody mark line is provided with an IgM antigen only bound with the IgM antibody, and the immunochromatographic region 43 on the left side of the IgG antibody mark line is provided with an IgG antigen only bound with the IgG antibody, so that if the corresponding antibody exists in the sample solution, the corresponding antibody is bound with the antigen of the corresponding immunochromatographic region, and meanwhile, the luminescent material is correspondingly attached to the corresponding immunochromatographic region 43.
By utilizing the time-resolved fluorescence immunochromatography system, the immunochromatography detection method can be completed through the following steps:
synchronously triggering a timing starting device of an excitation light emitting device and an image acquisition device, so that the timing starting device starts timing while the excitation light emitting device emits excitation light to a detected reagent strip;
until the preset time length is reached, the image acquisition device acquires images of the detected reagent strip after the optical filter, light in a specific wavelength range in light reflected by the detected reagent strip passes through the optical filter and then enters a lens of the image acquisition device, and light outside the specific wavelength range is filtered by the optical filter and cannot enter the lens of the image acquisition device;
judging whether a reference line area on the reagent strip is displayed or not, if so, continuing to execute the next step, and otherwise, judging that the detection result is invalid;
according to the showing condition of the corresponding immunochromatography area, obtaining an immunochromatography detection result, which comprises the following specific steps:
the specific manifestation principle is as described above, and the attached drawings 4-7 in the specification are four imaging results of the reagent strip: referring to fig. 4, only the fiducial marker line is shown, and neither IgM nor IgG-corresponding immunochromatographic zone 43 is shown, indicating that no IgM antibodies and IgG antibodies are present in the sample solution; referring to fig. 5, only the fiducial marker line and the immunochromatographic zone 43 corresponding to IgM are visualized, while the immunochromatographic zone 43 corresponding to IgG is not visualized, indicating that no IgG antibodies are present in the sample solution; referring to fig. 6, only the fiducial marker line and the immunochromatographic zone 43 corresponding to IgG are revealed, and the immunochromatographic zone 43 corresponding to IgM is not revealed, indicating that IgM antibodies are not present in the sample solution; referring to fig. 7, the fiducial marker lines and the immunochromatographic zones 43 for IgM and IgG, respectively, appear, indicating the presence of both IgM and IgG antibodies in the sample solution. If the reference mark line does not appear in the imaged image, the detection is invalid.
The identification of the immunochromatography detection result can be realized by manually identifying the acquired image, or automatically calibrating and identifying the acquired image through corresponding image identification software, and providing the identification result after detection for a user.
In searching similar patents, patent CN108627482A uses a special lighting and imaging device, the mobile terminal is only used as a carrier of the imaging device and analysis software, and the color of chromogen is used as the interpretation of the result, which is essentially different from the method used in this patent.
Patent CN 105842217 a also provides a detection method for photographing by mobile terminal, but this method still does not get rid of the conventional detection method, and unlike the conventional detection method, the method specially manufactures the light-emitting device, the optical path component, and the filtering component in the conventional detection into one device, and uses the mobile terminal to image and cooperate with the detection, which is completely different from the synchronous excitation lamp described in this patent.
The invention discloses a time-resolved fluorescence chromatography test detection method and a device based on mobile terminal photographing, wherein a specific exciting light device generates exciting light of lambda 1, the exciting light directly acts on a reaction film layer of a reagent card strip, a special light path design and related equipment are not needed, and meanwhile, a device which avoids the interference of ambient light such as a dark box is not needed based on a time-resolved fluorescence technology, and the time-resolved fluorescence chromatography test detection method and the device can be directly used under natural light.
The imaging device described in this patent is provided with a dedicated automatic detection software in addition to a general mobile terminal with a camera device, and when the imaging device is aligned with a card strip to be detected, the imaging device can automatically identify and quantitatively calibrate lambda 2 generated after excitation, and give a quantitative or qualitative detection result.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A shooting detection method based on time-resolved fluorescence tomography is characterized by comprising the following steps:
synchronously triggering a timing starting device of an excitation light emitting device and an image acquisition device, so that the timing starting device starts timing while the excitation light emitting device emits excitation light to a detected reagent strip;
until the preset time length is reached, the image acquisition device acquires images of the detected reagent strip after the optical filter, light in a specific wavelength range in light reflected by the detected reagent strip passes through the optical filter and then enters a lens of the image acquisition device, and light outside the specific wavelength range is filtered by the optical filter and cannot enter the lens of the image acquisition device.
2. The imaging detection method based on time-resolved fluorescence tomography according to claim 1, wherein the specific wavelength range is set according to a wavelength range of light emitted by excitation of a luminescent substance in the sample on the test reagent strip.
3. The photographing detection method based on time-resolved fluorescence tomography of claim 1, wherein the preset timing duration is set according to a time gap between excitation of a luminescent substance in a sample to be detected on the detected reagent strip and light emission and/or a decay time of the emitted light.
4. The photographing detection method based on the time-resolved fluorescence tomography of claim 1, wherein the timing starting device of the excitation light emitting device and the image acquisition device realizes synchronous triggering by the following methods:
starting a flash lamp of the image acquisition device;
when the flash information is detected, triggering the excitation light emitting device to emit excitation light to the detected reagent strip, and simultaneously triggering the timing starting device of the image acquisition device to start timing.
5. The photographing detection method based on the time-resolved fluorescence tomography of claim 1, wherein the timing starting device of the excitation light emitting device and the image acquisition device realizes synchronous triggering by the following methods:
and the same trigger signal generator is connected with the exciting light emitting device and the timing starting device in a wired mode, under the triggering action of the trigger signal generator, the exciting light emitting device emits exciting light, and meanwhile, the timing starting device starts timing.
6. A shooting detection device based on time-resolved fluorescence tomography is characterized by comprising an exciting light emitting device (1) for emitting exciting light, an optical filter (2) for filtering interference light, an image acquisition device (3) for acquiring image information of a detected reagent strip, and a synchronous trigger device for synchronizing the exciting light emitting device (1) with a timing starting device (31) of the image acquisition device (3), wherein the timing starting device (31) is used for starting the image acquisition device (3) to acquire images of the detected reagent strip (4) at regular time, the optical filter (2) is arranged between the detected reagent strip (4) and the image acquisition device (3), so that light in a specific wavelength range in the light reflected by the detected reagent strip (4) passes through the optical filter (2) and then enters a lens of the image acquisition device (3);
under the triggering action of the synchronous triggering device, the excitation light emitting device (1) emits excitation light to the detected reagent strip (4), and the timing starting device (31) starts to time;
and when the preset timing duration is reached, the image acquisition device (3) acquires the image of the detected reagent strip (4).
7. The shooting detection apparatus according to claim 6, characterized in that the synchronous trigger apparatus comprises a controller, a flash provided at the image capture apparatus (3), and a flash receiving apparatus (5) configured to detect flash information, the flash receiving apparatus (5), the excitation light emitting apparatus (1), the timing start apparatus (31) all being electrically connected with the controller;
when the flash light receiving device (5) detects flash light information, the controller controls to start the exciting light emitting device (1) to emit exciting light, and simultaneously controls the timing starting device (31) to start timing.
8. A time-resolved fluoroimmunoassay system comprising a reagent strip (4) and the imaging detection device based on time-resolved fluoroimmunoassay of claim 6 or 7, wherein the reagent strip (4) is provided with a sample inlet (41), a reference line region (42) and at least one immunochromatographic region (43), a sample solution fed through the sample inlet (41) is labeled with a specified luminescent substance, and the specific wavelength range filtered by the filter (2) of the imaging detection device based on time-resolved fluoroimmunoassay matches the wavelength range of light emitted by the specified luminescent substance when excited.
9. The time-resolved fluoroimmunoassay system according to claim 8, wherein the preset time duration of the timing starting means (31) of the time-resolved fluoroimmunoassay-based imaging detection means matches the time gap from excitation to emission of the specified luminescent substance and/or the decay time of the emitted light.
10. An immunochromatography detection method based on the time-resolved fluorescence immunochromatography system of claim 8 or 9, comprising the steps of:
synchronously triggering a timing starting device of an excitation light emitting device and an image acquisition device, so that the timing starting device starts timing while the excitation light emitting device emits excitation light to a detected reagent strip;
until the preset time length is reached, the image acquisition device acquires images of the detected reagent strip after the optical filter, light in a specific wavelength range in light reflected by the detected reagent strip passes through the optical filter and then enters a lens of the image acquisition device, and light outside the specific wavelength range is filtered by the optical filter and cannot enter the lens of the image acquisition device;
judging whether a reference line area on the reagent strip is displayed or not, if so, continuing to execute the next step, and otherwise, judging that the detection result is invalid;
and obtaining an immunochromatography detection result according to the appearance condition of the corresponding immunochromatography area.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112461806A (en) * | 2020-11-18 | 2021-03-09 | 厦门大学 | Fluorescence spectrum detection method based on smart phone |
| CN114441411A (en) * | 2021-12-31 | 2022-05-06 | 江苏汇先医药技术有限公司 | Method and system for interpreting capture result of tumor cell capture chip |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112461806A (en) * | 2020-11-18 | 2021-03-09 | 厦门大学 | Fluorescence spectrum detection method based on smart phone |
| CN114441411A (en) * | 2021-12-31 | 2022-05-06 | 江苏汇先医药技术有限公司 | Method and system for interpreting capture result of tumor cell capture chip |
| CN114441411B (en) * | 2021-12-31 | 2023-12-19 | 江苏汇先医药技术有限公司 | Method and system for interpreting capturing result of tumor cell capturing chip |
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