CN113125694B - Detection system for realizing classification and quantitative analysis and detection method for immune multi-joint detection - Google Patents
Detection system for realizing classification and quantitative analysis and detection method for immune multi-joint detection Download PDFInfo
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- CN113125694B CN113125694B CN201911420986.7A CN201911420986A CN113125694B CN 113125694 B CN113125694 B CN 113125694B CN 201911420986 A CN201911420986 A CN 201911420986A CN 113125694 B CN113125694 B CN 113125694B
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Abstract
The invention provides a detection system for realizing classification and quantitative analysis and a detection method for immune multi-joint detection, which comprise a sheath flow module, a first laser, a first fluorescence receiving module and a signal processing module, wherein the sheath flow module is used for enabling a plurality of magnetic sphere compounds to pass through a detection area of the sheath flow module one by one; the first laser is used for emitting first laser to the detection area so as to excite the first fluorescent substance to generate first fluorescence; the scattered light receiving module is used for receiving forward scattered light with N intensities corresponding to the standby magnetic balls with N particle sizes after passing through the detection area at the first side edge of the sheath flow module and converting the forward scattered light into N particle size type classification parameters; the first fluorescence receiving module is used for receiving the first fluorescence passing through the detection area at the second side of the sheath flow module and converting the first fluorescence into a counting parameter; the signal processing module is electrically connected with the scattered light receiving module and the first fluorescence receiving module, receives and processes the N particle size type classification parameters and the counting parameters so as to divide the multiple magnetic ball compounds into N types and count corresponding to each type.
Description
Technical Field
The invention relates to the technical field of blood cell analysis and detection, in particular to a detection system for realizing classification and quantitative analysis and a detection method for immune multi-joint detection.
Background
At present, in immunodiagnosis, a chemiluminescence immunoassay analyzer occupies a dominant position, and the technology utilizes a chemiluminescence mode to carry out qualitative and quantitative analysis on an object to be detected.
However, chemiluminescence has been problematic: only a single item can be detected in one detection process, the cost is high, the classification function is single, and when various items are detected, the detection processes need to be performed for multiple times.
Disclosure of Invention
The application provides a detection system for realizing classification and quantitative analysis and a detection method for immune multi-joint inspection, which aim to solve the technical problems that in the prior art, one detection process can only detect a single item, the cost is high, the classification function is single, and the time is consumed by multiple detection processes when multiple items are detected.
In order to solve the technical problem, the application adopts a technical scheme that: there is provided a detection system for performing classification and quantitative analysis, comprising:
the sheath flow module is used for enabling a plurality of magnetic sphere compounds to pass through a detection area of the sheath flow module one by one, wherein the plurality of magnetic sphere compounds comprise standby magnetic spheres with N particle size types, each standby magnetic sphere with each particle size is coupled with at least one substance to be detected, and the substance to be detected is combined with a first fluorescent substance;
a first laser for emitting a first laser to the detection region to excite the first fluorescent substance to generate a first fluorescence;
the scattered light receiving module is used for receiving forward scattered light with N intensities corresponding to the spare magnetic balls with N particle size types after passing through the detection area at the first side edge of the sheath flow module and converting the forward scattered light into N particle size type classification parameters;
a first fluorescence receiving module for receiving the first fluorescence passing through the detection area at the second side of the sheath flow module and converting the first fluorescence into a counting parameter;
and the signal processing module is electrically connected with the scattered light receiving module and the first fluorescence receiving module, and is used for receiving and processing the N particle size type classification parameters and the counting parameters so as to classify the multiple magnetic ball compounds into N classes and count the magnetic ball compounds corresponding to each class.
According to an embodiment of the present invention, each of the spare magnetic spheres has M excitation intensity levels of the second phosphor;
the detection system further comprises:
a second laser for emitting a second laser to the detection region to excite the second fluorescent substance with M intensity levels to generate a second fluorescence with M intensity levels;
a second fluorescence receiving module, configured to receive, at a second side of the sheath flow module, the M kinds of second fluorescence with intensity levels after passing through the detection area, and convert the M kinds of second fluorescence into M kinds of second fluorescence intensity classification parameters;
the signal processing module is also electrically connected with the second fluorescence receiving module and receives M second fluorescence intensity classification parameters so as to divide the multiple magnetic sphere compounds into N x M classes and count corresponding to each class.
According to an embodiment of the invention, the detection system further includes an optical filter disposed on a front side of the scattered light receiving module to allow the first laser light to pass through.
According to a specific embodiment of the present invention, the first laser and the second laser are beam splitting light, and are focused to the detection area through a lens group, and spots of the first laser and the second laser are overlapped in a horizontal direction and spaced in a vertical direction.
According to a specific embodiment of the present invention, the second laser and the first laser are combined beams, and the light spots of the first laser and the second laser are overlapped in the horizontal direction and spaced in the vertical direction;
the detection system further comprises a first dichroic mirror, the first laser emits first laser light towards a first direction, the second laser emits second laser light towards a second direction, and the first laser light and the second laser light form combined light through the first dichroic mirror; or
The detection system further comprises a first dichroic mirror and a first reflective mirror, the first laser emits first laser light towards the first direction, the second laser emits second laser light towards the first direction, and the first laser light and the second laser light form combined light through the first dichroic mirror and the first reflective mirror.
According to an embodiment of the present invention, the scattered light receiving module further includes a diaphragm, and the diaphragm is provided with a first light passing hole to receive the forward scattered light.
According to an embodiment of the present invention, the light receiving angle of the first light passing hole is 1 to 15 degrees.
According to an embodiment of the present invention, the diaphragm is further provided with a second light through hole at a side of the first light through hole, and the scattered light receiving module is further configured to receive, at the first side of the sheath flow module, the laterally scattered light that has passed through the detection area through the second light through hole, convert the laterally scattered light into an auxiliary classification parameter, and provide the auxiliary classification parameter to the signal processing module.
According to an embodiment of the present invention, the light receiving angle of the second light passing hole is 8 to 50 degrees.
According to a specific embodiment of the present invention, the detection system further comprises a second dichroic mirror for splitting the first fluorescence and the second fluorescence at a second side of the sheath flow module.
In order to solve the technical problem, the other technical scheme adopted by the application is as follows: the detection method for realizing the immune multi-joint detection is characterized by comprising the following steps of:
configuring a substance to be detected and a first fluorescent substance for standby magnetic balls with N grain size types to form a plurality of magnetic ball compounds;
performing optical detection on the magnetic sphere compound by combining a sheath flow module with first laser to obtain forward scattered light with N intensities corresponding to the standby magnetic spheres with N grain size types to serve as N grain size type classification parameters;
and performing fluorescence detection on the magnetic sphere compounds by combining a sheath flow module with first laser to obtain first fluorescence intensity corresponding to the first fluorescent substance as a counting parameter, and further dividing the plurality of magnetic sphere compounds into N types and counting the N types correspondingly.
The beneficial effect of this application is: different from the situation of the prior art, the detection system for realizing classification and quantitative analysis provided by the invention can perform N types of classification on various magnetic ball compounds by setting N types of particle sizes of the standby magnetic balls and detecting forward scattered light intensity, can perform M types of classification on various magnetic ball compounds by configuring M types of second fluorescent substances with excitation intensity levels on the standby magnetic balls and detecting corresponding fluorescence intensity, can be used independently or in combination, and can classify various magnetic ball compounds into N-M types by passing the various magnetic ball compounds through a sheath flow module once when the two types of classification are used in combination, so that the classification performance is greatly improved, the detection time is shortened, and the concentration of a substance to be detected on the magnetic ball compounds can be recorded through the first fluorescence intensity detection, thereby realizing the functions of rapid multi-classification and counting.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, it is obvious that the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without inventive efforts, wherein:
FIG. 1 is a simplified optical schematic of a detection system for performing classification and quantitative analysis provided by an embodiment of the present invention;
fig. 2 is a schematic plan view of a diaphragm of the scattered light receiving module shown in fig. 1.
FIG. 3 is a schematic illustration of various magnetic sphere composites of FIG. 2.
Detailed Description
The technical solutions in the embodiments of the present application will be described clearly and completely with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only some embodiments of the present application, and not all embodiments. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments in the present application without making any creative effort belong to the protection scope of the present application.
It should be noted that, if directional indications (such as up, down, left, right, front, back, 8230; etc.) are involved in the embodiment of the present invention, the directional indications are only used for explaining the relative positional relationship between the components, the motion situation, etc. in a specific posture (as shown in the figure), and if the specific posture is changed, the directional indications are correspondingly changed.
In addition, if there is a description relating to "first", "second", etc. in the embodiments of the present invention, the description of "first", "second", etc. is for descriptive purposes only and is not to be construed as indicating or implying relative importance or to implicitly indicate the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present invention.
The present embodiment provides a detection system for implementing classification and quantitative analysis, which includes a sheath flow module 100, a first laser 110, a second laser 120, a scattered light receiving module 130, a first fluorescence receiving module 140, a second fluorescence receiving module 150, and a signal processing module 160, wherein the scattered light receiving module 130, the first fluorescence receiving module 140, and the second fluorescence receiving module 150 are electrically connected to the signal processing module 160.
The sheath flow module 100 is configured to enable a plurality of magnetic sphere compounds to pass through a detection area of the sheath flow module 100 one by one, wherein the plurality of magnetic sphere compounds includes N types of particle sizes of spare magnetic spheres, each of the spare magnetic spheres is coupled with at least one substance to be detected, and the substance to be detected is combined with a first fluorescent substance.
The substance to be detected may be a plurality of antigens in a blood sample, such as antigens A/B/C/D/D/E/F/G/H/I, or other substances requiring classification and counting.
Wherein, when N =3, namely the particle size of the spare magnetic ball has three different particle size types Φ 1, Φ 2, Φ 3, the particle size may be 1um, 4um, 7um, or 2um, 5um, 8um, or 3um, 6um, 9um, etc., and the different spare magnetic balls are coupled with the corresponding antigen a/B/C/D/E/F/G/H/I in the blood sample through the corresponding antibody a/B/C/D/E/F/G/H/I.
The first fluorescent substance can be a green fluorescent substance or other substances with similar labeling functions, and the substance to be detected is bound to the first fluorescent substance through the antibody r.
The first laser 110 is used for emitting a first laser to the detection region to excite the first phosphor to generate a first fluorescence, and the first laser 110 may be a green laser with a wavelength of 532nm.
The scattered light receiving module 130 is configured to receive forward scattered light of N intensities corresponding to the N types of particle size type spare magnetic spheres after passing through the detection area at the first side of the sheath flow module 100 and convert the forward scattered light into N types of particle size type classification parameters. The scattered light receiving module 130 includes conventional optical elements such as a diaphragm 133, a lens, a filter, a signal detector, and the like.
When the spare magnetic spheres have 3 types of particle sizes, a variety of magnetic sphere compounds can be classified into 3 types by forward scattered light signals corresponding to 3 intensities.
When the spare magnetic spheres have 5 types of particle sizes, a variety of magnetic sphere compounds can be classified into 5 types by forward scattered light signals corresponding to 5 intensities.
The first fluorescence receiving module 140 is configured to receive the first fluorescence passing through the detection region at the second side of the sheath flow module 100 and convert the first fluorescence into a count parameter, where each magnetic sphere compound corresponds to one first fluorescence signal, and the concentration of the analyte on the magnetic sphere compound can be calculated according to the intensity of the first fluorescence signal, so as to implement quantitative analysis. The first fluorescence receiving module 140 may include a collecting lens group, a collimating lens group, a focusing lens group, a band pass filter, a fluorescence detector, and other optical elements, where the collimating lens group may be an aspheric lens or a plurality of lenses for collimating the fluorescence into parallel light or approximately parallel light, and the focusing lens group may be an aspheric lens or a plurality of lenses for focusing the fluorescence on the fluorescence detector.
Preferably, the fluorescence detector may be selected from a photomultiplier tube (PMT).
Alternatively, the fluorescence detector may be a silicon photomultiplier (SiPM, MPPC).
Alternatively, the fluorescence detector may be an Avalanche Photo Diode (APD).
Alternatively, the fluorescence detector may be selected from a Photodiode (PD).
The signal processing module 160 receives and processes the N particle size type classification parameters and the counting parameters, thereby classifying the plurality of magnetic ball compounds into N classes and counting the N classes.
Further, the detection system provided by the embodiment of the present invention further has a stronger classification function, and accordingly, each of the spare magnetic spheres of the particle size type has M excitation intensity levels of the second fluorescent substance, wherein when M =3, that is, the excitation intensity level of the second fluorescent substance of the spare magnetic sphere is 3 levels, for example, three spare magnetic spheres of the same particle size in fig. 3 have colorless fills, 1/4 fills, and 1/2 fills corresponding to the second fluorescent substance configuration representing three excitation intensity levels.
The second laser 120 is used for emitting a second laser to the detection area to excite the second phosphor with M intensity levels to generate a second fluorescence with M intensity levels, and the second laser 120 may be a red laser with a wavelength of 638nm.
The second fluorescence receiving module 150 is configured to receive the M kinds of second fluorescence with intensity levels after passing through the detection region at the second side of the sheath flow module 100 and convert the M kinds of second fluorescence into M kinds of second fluorescence intensity classification parameters. Similarly, the second fluorescence receiving module 150 may include a collecting lens group, a collimating lens group, a focusing lens group, a band pass filter, a fluorescence detector, and other optical elements, where the collimating lens group may be an aspheric lens or a plurality of lenses for collimating the fluorescence into parallel light or approximately parallel light, and the focusing lens group may be an aspheric lens or a plurality of lenses for focusing the fluorescence on the fluorescence detector.
Preferably, the fluorescence detector may be selected from a photomultiplier tube (PMT).
Alternatively, the fluorescence detector may be a silicon photomultiplier (SiPM, MPPC).
Alternatively, the fluorescence detector may be an Avalanche Photo Diode (APD).
Alternatively, the fluorescence detector may be selected from a Photodiode (PD)
When the excitation intensity level of the second fluorescent substance of the spare magnetic sphere is 3 levels, the plurality of magnetic sphere complexes can be classified into 3 classes by the second fluorescent signal corresponding to the 3 intensities.
When the excitation intensity level of the second fluorescent substance of the spare magnetic sphere is 5 levels, the plurality of magnetic sphere complexes can be classified into 5 classes by the second fluorescent signal corresponding to the 5 intensities. The signal processing module 160 further classifies the plurality of magnetic sphere complexes into N × M classes according to the M second fluorescence intensity classification parameters and counts the number of each class.
Specifically, the detection system further includes an optical filter disposed on a front side of the scattered light receiving module 130 to block the interference light and allow the first laser light to pass through.
In the embodiment of the invention, the first laser and the second laser can be beam splitting light, the beam splitting light is focused to the detection area through the lens group, the light spots of the first laser and the second laser are overlapped in the horizontal direction, and the light spots are spaced in the vertical direction to avoid the mutual interference of two paths of fluorescence.
In the embodiment of the invention, the first laser and the second laser can be beam combination light, and light spots of the first laser and the second laser are overlapped in the horizontal direction and are spaced in the vertical direction to avoid mutual interference of two paths of fluorescence.
Specifically, when the first laser and the second laser may be combined light, the detection system further includes a first dichroic mirror 115, the first laser 110 emits first laser light toward the first direction, the second laser 120 emits second laser light toward the second direction, the first laser and the second laser form combined light through the first dichroic mirror 115, and the first direction and the second direction are perpendicular to each other. Alternatively, the detection system further includes a first dichroic mirror 115 and a first reflective mirror, the first laser 110 emits first laser light toward the first direction, the second laser 120 emits second laser light toward the first direction, and the first laser light and the second laser light form a combined light through the first dichroic mirror 115 and the first reflective mirror. The first laser 110 and the second laser 120 are arranged in parallel and spaced apart, so that the structure of the detection system can be more compact.
As shown in fig. 2, in the embodiment of the present invention, the diaphragm 133 is provided with a first light passing hole 131 to receive the forward scattered light, the first light passing hole 131 may be a sector area arranged around, and avoid the over-strong forward scattered light at the center, for example, 4 sector areas, and the light receiving angle of the first light passing hole 131 may be 1-15 degrees.
The diaphragm 133 is further provided with a second light through hole 132 on a side of the first light through hole 131, the scattered light receiving module 130 is further configured to receive, at the first side of the sheath flow module 100, the laterally scattered light that passes through the detection area through the second light through hole 132, convert the laterally scattered light into an auxiliary classification parameter, and provide the auxiliary classification parameter to the signal processing module 160, the type of the spare magnetic ball, such as the particle size type of the spare magnetic ball or the internal structure type of the spare magnetic ball, can be distinguished through the laterally scattered light, and the light receiving angle of the second light through hole 132 is 8 to 50 degrees.
In an embodiment of the present invention, the detection system further comprises a second dichroic mirror 145, and the second dichroic mirror 145 is configured to split the first fluorescence and the second fluorescence at the second side of the sheath flow module 100.
In the embodiment, the standby magnetic balls are set to be N-type in particle size, the multiple magnetic ball compounds can be classified into N types through forward scattered light intensity detection, the standby magnetic balls are provided with M excitation intensity levels of the second fluorescent substances, the multiple magnetic ball compounds can be classified into M types through corresponding fluorescence intensity detection, the two types of classification modes can be used independently or in combination, when the two types of classification modes are combined for use, the multiple magnetic ball compounds can be classified into N M types through the sheath flow module at one time, the classification performance is greatly improved, the detection time is shortened, the concentration of the to-be-detected object on the magnetic ball compounds can be recorded through the first fluorescence intensity detection, and therefore the functions of rapid multi-classification and counting are achieved.
The detection system for realizing classification and quantitative analysis provided by the invention has the advantages of simple structure and lower cost, and can realize rapid multi-classification and counting functions only by utilizing two lasers, two fluorescence receiving modules and one quantitative fluorescent reagent.
The invention also provides a detection method for realizing the immune multi-joint detection, which specifically comprises the following steps:
configuring a substance to be detected and a first fluorescent substance for standby magnetic balls with N grain size types to form a plurality of magnetic ball compounds;
performing optical detection on the magnetic sphere compound by combining a sheath flow module with first laser to obtain N forward scattering light corresponding to the standby magnetic spheres of N particle size types to serve as N particle size type classification parameters;
and performing fluorescence detection on the magnetic sphere compounds by combining a sheath flow module with the first laser to obtain first fluorescence intensity corresponding to the first fluorescent substance as a counting parameter, so that the multiple magnetic sphere compounds are divided into N types and each type is counted.
The multiple magnetic ball compounds comprise spare magnetic balls with N grain sizes, each spare magnetic ball with each grain size is coupled with at least one substance to be detected, and the substance to be detected is combined with a first fluorescent substance.
The substance to be detected may be a plurality of antigens in a blood sample, such as antigens A/B/C/D/D/E/F/G/H/I, or other substances requiring classification and counting.
Wherein, when N =3, namely the particle size of the spare magnetic ball has three different particle size types of Φ 1, Φ 2, Φ 3, the particle size can be specifically 1um, 4um, 7um, or 2um, 5um, 8um, or 3um, 6um, 9um, etc., and the different spare magnetic balls are coupled with the corresponding antigen a/B/C/D/E/F/G/H/I in the blood sample through the corresponding antibody a/B/C/D/E/F/G/H/I.
The first fluorescent substance can be a green fluorescent substance or other substances with similar labeling functions, and the substance to be detected is bound to the first fluorescent substance through the antibody r.
The first laser is used for exciting the first fluorescent substance to generate first fluorescence, and the first laser can be green laser with the wavelength of 532nm.
The spare magnetic balls with N grain size types can correspondingly generate forward scattered light with N intensities when being irradiated by the first laser, and the forward scattered light serves as N grain size type classification parameters.
When the spare magnetic spheres have 3 types of particle sizes, a variety of magnetic sphere compounds can be classified into 3 types by forward scattered light signals corresponding to 3 intensities.
When the spare magnetic spheres have 5 types of particle sizes, a variety of magnetic sphere compounds can be classified into 5 types by forward scattered light signals corresponding to 5 intensities.
Each magnetic ball compound corresponds to a first fluorescence signal, and the concentration of the object to be detected on the magnetic ball compound can be calculated through the intensity of the first fluorescence signal, so that quantitative analysis is realized.
Further, the detection method provided by the embodiment of the invention also has a stronger classification function.
Specifically, M kinds of second fluorescent substances of excitation intensity levels are arranged for each particle size type of spare magnetic balls.
When M =3, that is, the excitation intensity level of the second fluorescent substance of the spare magnetic ball is 3 levels, for example, three spare magnetic balls with the same particle size in fig. 3 have colorless fills, 1/4 fills, and 1/2 fills corresponding to the second fluorescent substance arrangement representing three excitation intensity levels.
And performing fluorescence detection on the magnetic sphere compound by combining a sheath flow module with second laser to obtain M second fluorescence with the M intensity levels corresponding to the M second fluorescent substances and using the M second fluorescence as M second fluorescence intensity classification parameters. The second laser can be red laser with wavelength of 638nm.
Combining the classification parameters of N types of particle sizes and M types of fluorescence intensities, the magnetic ball compounds can be classified into N × M types and counted corresponding to each type.
The detection method comprises the following steps:
and the interference light is blocked by using the optical filter to obtain the forward scattered light of the first laser so as to reduce the interference.
In the detection method, the first laser and the second laser which are split into beams can be focused to a detection area of the sheath flow module through the lens group, light spots of the first laser and the second laser are overlapped in the horizontal direction and are spaced in the vertical direction, and mutual interference of two paths of fluorescence is avoided.
In the detection method, the first laser and the second laser can be combined to enable light spots of the first laser and the second laser to be overlapped in the horizontal direction, and the first laser and the second laser are spaced in the vertical direction to avoid mutual interference of two paths of fluorescence.
Specifically, the first laser light and the second laser light may be formed into combined light using the first dichroic mirror 115; or the first laser light and the second laser light are formed into a combined light by using the first dichroic mirror 115 and the first reflective mirror.
As shown in fig. 3, in order to accurately identify the forward scattered light, a diaphragm 133 provided with a first light passing hole 131 may be used to receive the forward scattered light passing through the detection region to receive the forward scattered light of 1-15 degrees. The side scattered light passing through the detection region may also be received by a diaphragm 133 provided with a second light passing hole 132 to receive 8-50 degrees of side scattered light, which may be used to distinguish the type of spare magnetic sphere. In the detection of the two fluorescence, the first fluorescence and the second fluorescence can be split by using the second dichroic mirror 145.
In the embodiment, the standby magnetic balls are set to be of N types of particle sizes, the multiple magnetic ball compounds can be classified into N types through forward scattered light intensity detection, the standby magnetic balls are provided with M types of second fluorescent substances with excitation intensity levels, the multiple magnetic ball compounds can be classified into M types through corresponding fluorescence intensity detection, the two classification modes can be used independently or in combination, when the two classification modes are used in combination, the multiple magnetic ball compounds can be classified into N M types through the sheath flow module at one time, the classification performance is greatly improved, the detection time is shortened, the concentration of an object to be detected on the magnetic ball compounds can be recorded through the first fluorescence intensity detection, and therefore the rapid multi-classification and counting functions are achieved.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by using the contents of the present specification and the accompanying drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (18)
1. A detection system for performing classification and quantitative analysis, comprising:
the sheath flow module is used for enabling a plurality of magnetic sphere compounds to pass through a detection area of the sheath flow module one by one, wherein the plurality of magnetic sphere compounds comprise standby magnetic spheres with N particle size types, each standby magnetic sphere with each particle size is coupled with at least one substance to be detected, and the substance to be detected is combined with a first fluorescent substance;
a first laser for emitting a first laser to the detection region to excite the first fluorescent substance to generate a first fluorescence;
the scattered light receiving module is used for receiving forward scattered light with N intensities corresponding to the spare magnetic balls with N particle size types after passing through the detection area at the first side edge of the sheath flow module and converting the forward scattered light into N particle size type classification parameters;
a first fluorescence receiving module for receiving the first fluorescence passing through the detection area at the second side of the sheath flow module and converting the first fluorescence into a counting parameter;
the signal processing module is electrically connected with the scattered light receiving module and the first fluorescence receiving module, and is used for receiving and processing the N particle size type classification parameters and the counting parameters so as to divide the plurality of magnetic ball compounds into N classes and count the magnetic ball compounds corresponding to each class;
each particle size type of the spare magnetic ball is provided with M excitation intensity levels of second fluorescent substances, and the laser intensity levels are configured according to the filling mode of the spare magnetic ball;
the detection system further comprises:
a second laser for emitting a second laser beam to the detection region to excite the second fluorescent substance with M intensity levels to generate a second fluorescence with M intensity levels;
a second fluorescence receiving module, configured to receive, at a second side of the sheath flow module, the M kinds of second fluorescence with intensity levels after passing through the detection area, and convert the M kinds of second fluorescence into M kinds of second fluorescence intensity classification parameters;
the signal processing module is also electrically connected with the second fluorescence receiving module and receives M second fluorescence intensity classification parameters so as to divide the multiple magnetic sphere compounds into N x M classes and count corresponding to each class;
the first laser emits first laser light towards a first direction, the second laser emits second laser light towards a second direction, and the first direction and the second direction are mutually perpendicular;
the light spots of the first laser and the second laser are overlapped in the horizontal direction and are spaced in the vertical direction.
2. The detection system for enabling classification and quantitative analysis according to claim 1, wherein:
the detection system further comprises an optical filter, and the optical filter is arranged on the front side of the scattered light receiving module to allow the first laser to pass through.
3. The detection system for classification and quantitative analysis according to claim 1, wherein:
the first laser and the second laser are beam splitting light and are focused to the detection area through a lens group.
4. The detection system for classification and quantitative analysis according to claim 1, wherein:
the second laser and the first laser are beam combination light, and light spots of the first laser and the second laser are overlapped in the horizontal direction and spaced in the vertical direction;
the detection system further comprises a first dichroic mirror, the first laser emits first laser light towards the first direction, the second laser emits second laser light towards the second direction, and the first laser light and the second laser light form combined light through the first dichroic mirror.
5. The detection system for realizing classification and quantitative analysis according to claim 1, wherein the scattered light receiving module further comprises a diaphragm, and the diaphragm is provided with a first light through hole to receive the forward scattered light.
6. The detection system for realizing classification and quantitative analysis according to claim 5, wherein the light receiving angle of the first light passing hole is 1-15 degrees.
7. The detection system for realizing classification and quantitative analysis according to claim 5, wherein the diaphragm is further provided with a second light through hole at a side of the first light through hole, and the scattered light receiving module is further configured to receive laterally scattered light passing through the detection area at the first side of the sheath flow module through the second light through hole, and convert the laterally scattered light into auxiliary classification parameters to be provided to the signal processing module.
8. The detection system for realizing classification and quantitative analysis according to claim 7, wherein the light receiving angle of the second light passing hole is 8-50 degrees.
9. The detection system for enabling classification and quantitative analysis according to claim 1, further comprising a second dichroic mirror for splitting the first and second fluorescence on a second side of the sheath flow module.
10. A detection method for realizing immune multi-joint detection is characterized by comprising the following steps:
configuring a substance to be detected and a first fluorescent substance for standby magnetic balls with N grain size types to form a plurality of magnetic ball compounds;
performing optical detection on the magnetic sphere compound by combining a sheath flow module with first laser to obtain forward scattered light with N intensities corresponding to the standby magnetic spheres with N grain size types to serve as N grain size type classification parameters;
performing fluorescence detection on the magnetic sphere compound by combining a sheath flow module with first laser to obtain first fluorescence intensity corresponding to the first fluorescent substance as a counting parameter, and further dividing the plurality of magnetic sphere compounds into N types and counting the N types corresponding to each type; the detection method further comprises the following steps:
configuring M second fluorescent substances with excitation intensity levels for the spare magnetic spheres of each particle size type, wherein the laser intensity levels are configured according to the filling mode of the spare magnetic spheres;
performing fluorescence detection on the magnetic sphere compound by combining a sheath flow module with second laser to obtain M kinds of second fluorescence with M kinds of excitation intensity levels, wherein the M kinds of second fluorescence are corresponding to the second fluorescent substance and serve as M kinds of second fluorescence intensity classification parameters;
combining the N particle size type classification parameters and the M fluorescence intensity classification parameters to classify the multiple magnetic sphere compounds into N × M classes and counting the number of each class;
the first laser emits first laser light towards a first direction, the second laser emits second laser light towards a second direction, and the first direction and the second direction are perpendicular to each other;
the light spots of the first laser and the second laser are overlapped in the horizontal direction and are spaced in the vertical direction.
11. The assay method for performing immune multiplexed assay according to claim 10, wherein the assay method further comprises:
and blocking interference light by using an optical filter to obtain forward scattered light of the first laser.
12. The detection method for realizing immune multi-joint detection according to claim 10, wherein:
the first laser and the second laser which are split are focused to a detection area of a sheath flow module through a lens group, and light spots of the first laser and the second laser are overlapped in the horizontal direction and are spaced in the vertical direction.
13. The detection method for realizing immune multi-joint detection according to claim 10, wherein:
combining the first laser and the second laser to enable light spots of the first laser and the second laser to be overlapped in the horizontal direction and spaced in the vertical direction;
forming the first laser light and the second laser light into combined light by using a first dichroic mirror; or
And forming the first laser and the second laser into combined light by using a first dichroic mirror and a first reflective mirror.
14. The detection method for realizing immune multi-joint detection according to claim 10, wherein the forward scattered light is received by a diaphragm provided with a first light passing hole.
15. The detection method for realizing immune multi-joint detection according to claim 10, wherein the forward scattered light is received at 1-15 degrees.
16. The detection method for realizing immune multi-joint detection according to claim 14, wherein the diaphragm provided with a second light through hole is used for receiving side scattered light.
17. The detection method for realizing immune multi-joint detection according to claim 16, wherein the side scattered light is received at 8-50 degrees.
18. The detection method for realizing immune multiplexed assay according to claim 10, wherein the first fluorescence and the second fluorescence are received after being split by a second dichroic mirror.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103735249A (en) * | 2013-12-27 | 2014-04-23 | 中国科学院苏州生物医学工程技术研究所 | Fluorescence detector |
| CN108204960A (en) * | 2016-12-16 | 2018-06-26 | 深圳市帝迈生物技术有限公司 | A kind of analysis system and method for coding microball |
| CN208155810U (en) * | 2018-04-24 | 2018-11-27 | 中翰盛泰生物技术股份有限公司 | A kind of optical system and device for streaming phosphor collection |
| CN110514575A (en) * | 2019-08-16 | 2019-11-29 | 迈克医疗电子有限公司 | The decoding apparatus and method of quantum point coding microball |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101441212A (en) * | 2008-12-04 | 2009-05-27 | 上海交通大学 | Multiple-antigen synchronous detection method of quantum dot mark fluorescent immune |
| JP6100658B2 (en) * | 2013-03-29 | 2017-03-22 | シスメックス株式会社 | Blood cell analyzer and blood cell analysis method |
| US9513206B2 (en) * | 2013-03-29 | 2016-12-06 | Sysmex Corporation | Particle measuring apparatus |
| CN203337547U (en) * | 2013-06-27 | 2013-12-11 | 中国科学院苏州生物医学工程技术研究所 | Compact type single-light-source multi-channel flow type analyzer |
| CN105021578B (en) * | 2014-04-15 | 2019-04-19 | 杭州微瑞科技有限公司 | Fluid fluorescent quantitative detection device and method |
| CN105784662A (en) * | 2016-04-27 | 2016-07-20 | 武汉大学 | Liquid-phase suspension biochip based on multi-optical trap encoding bead array and two-photon fluorescence detection |
| CN205691502U (en) * | 2016-06-01 | 2016-11-16 | 章健 | A kind of magnetic microsphere fluorescence detecting system |
| CN108072637B (en) * | 2016-11-15 | 2023-05-26 | 深圳市帝迈生物技术有限公司 | Flow quantum dot blood multi-component analysis system and analysis method |
| CN107561043B (en) * | 2017-07-06 | 2020-05-12 | 复旦大学 | Biosensor based on multicolor up-conversion coding fluorescence technology |
| CN108132208A (en) * | 2017-12-25 | 2018-06-08 | 黄庆 | A kind of spiral shape microchannel and its application method and series and parallel installation method |
| US11040347B2 (en) * | 2018-06-14 | 2021-06-22 | Owl biomedical, Inc. | Microfabricated droplet dispensor with immiscible fluid |
| CN110361370B (en) * | 2019-07-31 | 2022-03-08 | 深圳中山泌尿外科医院 | Single-embryo secretory protein quantitative detection method based on microfluidic droplets |
-
2019
- 2019-12-31 CN CN201911420986.7A patent/CN113125694B/en active Active
-
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- 2020-12-18 WO PCT/CN2020/137727 patent/WO2021135991A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103735249A (en) * | 2013-12-27 | 2014-04-23 | 中国科学院苏州生物医学工程技术研究所 | Fluorescence detector |
| CN108204960A (en) * | 2016-12-16 | 2018-06-26 | 深圳市帝迈生物技术有限公司 | A kind of analysis system and method for coding microball |
| CN208155810U (en) * | 2018-04-24 | 2018-11-27 | 中翰盛泰生物技术股份有限公司 | A kind of optical system and device for streaming phosphor collection |
| CN110514575A (en) * | 2019-08-16 | 2019-11-29 | 迈克医疗电子有限公司 | The decoding apparatus and method of quantum point coding microball |
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