CN113720825A - Optical instant detector and detection method and application - Google Patents
Optical instant detector and detection method and application Download PDFInfo
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- CN113720825A CN113720825A CN202111297412.2A CN202111297412A CN113720825A CN 113720825 A CN113720825 A CN 113720825A CN 202111297412 A CN202111297412 A CN 202111297412A CN 113720825 A CN113720825 A CN 113720825A
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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
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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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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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
- G01N2021/6463—Optics
Abstract
The embodiment of the invention provides an optical instant detector, a detection method and application, wherein the detector comprises: the device comprises a light source emitter, a Y-shaped optical fiber bundle, a fluorescence intensity detector and a reflector; the Y-shaped optical fiber bundle is provided with an incident optical fiber section, an emergent optical fiber section and a public section which is simultaneously connected with the incident optical fiber section and the emergent optical fiber section, the incident optical fiber section is provided with an incident end face, the light source emitter faces the incident end face, the emergent optical fiber section is provided with an emergent end face, the emergent end face faces the fluorescence intensity detector, and the public section is provided with a fluorescence receiving end face which is used for transmitting light rays emitted by the light source emitter and can receive fluorescence. When the light intensity is calibrated, the reflecting mirror is arranged opposite to the fluorescence receiving end face. The detection method comprises the following steps: the optical instant detector provided by the application is used for detecting the concentration of the sample. The detector can realize light intensity calibration by utilizing the reflector without additionally arranging a light path as in the prior art, has few optical elements, simple structure and quick and convenient detection method, and is very suitable for being applied to medical inspection.
Description
Technical Field
The invention relates to the technical field of instant detection, in particular to an optical instant detector, a detection method and application.
Background
Point of care Testing (POCT), a "Point of care test" performed spatially on-site at a patient; the "point-of-care" test performed at the time of patient onset is understood temporally. POCT is a new method which can save the complex processing procedure of the sample in laboratory test and can quickly obtain the test result; or a movable miniature detection system which is arranged outside a central laboratory and is close to a detection object and can report the result in time.
Although the current POCT products can basically meet the demand of immediate and immediate use, the current POCT products have a large gap with the characteristics of 'portability' and 'miniaturity', which greatly limits the application scenes of the POCT products. One of the main reasons why current products cannot be portable is that: a free-space type optical system is used.
A conventional optical system for POCT is shown in fig. 1:
the sample container 1 is a sample container to be measured and is composed of a bottom plate 11 and a cover plate 12, wherein the cover plate 12 has a certain optical transmittance for an illumination spectrum, and contains a sample 101 to be measured at a fixed position in the sample container 1. The reference numeral 2 denotes an optical system of an optical sensor, and the light emitted from the light source 201 passes through an incident lens 202, a beam splitter 203, a dichroic mirror 206, a collecting lens 207, and a sample container cover 12 in sequence, and is irradiated onto the sample 101 to be measured. The sample 101 to be measured has a fluorescent substance, which excites fluorescence under the irradiation of light, the fluorescence is collected on the first photodetector 209 through the collecting lens 207, the dichroic mirror 206 and the first condenser lens 208, and the amount of the excited fluorescence in the sample to be measured is determined according to the intensity of the first photodetector 209, so as to determine the amount of the target substance connected to the fluorescent substance. Since there is a different spectral range between the excited fluorescence and the illumination light, the dichroic mirror functions to reflect the illumination light emitted from the light source onto the collection lens 207 and allow the fluorescence emitted from the sample to be measured to pass through it to the first condenser lens 208. In order to correct the deviation of the illumination intensity in real time, a reference light path is added at the illumination end and consists of a beam splitter 203, a second beam splitter lens 204 and a second photoelectric detector 205, a part of illumination light is detected on the second photoelectric detector 205 during each measurement, the intensity of the light emitted by the light source 201 is monitored in real time by using the part of the intensity, and if the deviation occurs, the intensity of the light source 201 is adjusted, or a coefficient is used for correcting the whole system.
Although the above optical path structure can effectively detect the amount of the target substance in the sample to be detected, the above optical path structure has the following disadvantages: 1) the structure is complex, and the portability of POCT detection equipment is a great obstacle; 2) the number of optical devices is large, and the cost is high.
Disclosure of Invention
The present invention is directed to an optical on-the-fly detector, a detection method and an application thereof, which are directed to overcoming at least one of the problems set forth in the background.
Embodiments of the invention may be implemented as follows:
in a first aspect, the present invention provides an optical instantaneous detector comprising:
the device comprises a light source emitter, a Y-shaped optical fiber bundle, a fluorescence intensity detector and a reflector.
The Y-shaped optical fiber bundle is provided with an incident optical fiber section, an emergent optical fiber section and a public section which is simultaneously connected with the incident optical fiber section and the emergent optical fiber section, the incident optical fiber section is provided with an incident end face, the light source emitter faces the incident end face, the emergent optical fiber section is provided with an emergent end face, the emergent end face faces the fluorescence intensity detector, and the public section is provided with a fluorescence receiving end face which is used for transmitting light rays emitted by the light source emitter and can receive fluorescence.
When the light intensity is calibrated, the reflecting mirror is arranged opposite to the fluorescence receiving end face.
In an alternative embodiment, a fiber coupling lens for coupling the emitted light into the Y-shaped fiber bundle is disposed between the light source emitter and the Y-shaped fiber bundle.
In an alternative embodiment, a focusing lens for focusing the emergent fluorescence to be captured by the fluorescence intensity detector is arranged between the Y-shaped fiber bundle and the fluorescence intensity detector.
In an alternative embodiment, the fluorescence detection device further comprises a sample container which is arranged opposite to the fluorescence receiving end face in use, wherein the sample container comprises a bottom plate and a cover plate with light permeability.
In a second aspect, the present invention provides a method for real-time sample detection, which uses an optical real-time detector as any one of the above, including:
light intensity calibration: will be provided withThe reflector is arranged at the position opposite to the fluorescence receiving end surface for light intensity calibration, and the fluorescence intensity detector is used for measuring the reference light intensity
According to the formula
The light intensity calibration factor is calculated,
setting a light intensity value for the factory of the light source emitter, wherein alpha is a light intensity calibration coefficient;
sample detection: and (3) placing the sample at a position opposite to the fluorescence receiving end surface to detect the fluorescence intensity of the sample, and calculating to obtain the actual concentration of the sample according to the measured fluorescence intensity, the light intensity calibration coefficient and the standard curve relation between the fluorescence intensity and the concentration of the sample to be detected.
In an alternative embodiment, the step of detecting the sample specifically comprises:
placing the sample to be detected at a position opposite to the fluorescence receiving end face, and measuring the fluorescence intensity of the fluorescent substance in the sample to be detected, which is irradiated by the light source and stimulated to emit through the fluorescence intensity detector
Calculating to obtain the target test concentration according to the standard curve
According to the formula
Obtaining the actual concentration
。
In an optional embodiment, the detecting further comprises: and detecting a plurality of standard samples with known concentrations by adopting an optical instant detector to obtain a plurality of fluorescence intensity values, and determining a standard curve of the fluorescence intensity and the concentration of the sample to be detected according to the plurality of known concentrations and the corresponding plurality of fluorescence intensity values.
In a third aspect, the present invention provides the use of the instant sample detection method of the previous embodiment in medical testing.
The beneficial effects of the embodiment of the invention include, for example:
the application provides an instant detector of optics, owing to reasonable structure setting, set up the speculum and can carry out the light path calibration in the position relative with fluorescence receiving end face, need not set up a light path alone like the current detector mentioned in the background art. The detector has a simple structure, parts are obviously less than those of the existing detectors, and the detector can be integrated into equipment which has a small volume and is convenient to take and use; because of the few optical parts, the manufacturing cost is lower.
The sample instant detection method implemented by the optical instant detector is simple, rapid and convenient to operate, and is very suitable for being applied to medical examination.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a diagram of an optical system of a conventional sample detection device;
FIG. 2 is a diagram of an optical system of the optical instant detector provided in the present embodiment;
FIG. 3 is a schematic diagram of the optical real-time detector during calibration;
FIG. 4 is a schematic diagram of an optical instantaneous detector during testing.
Icon: 1-a sample container; 11-a base plate; 12-a cover plate; 101-a sample to be detected; 4-an object stage; 401-a working position; 201-a light source; 202-an incident lens; 203-a spectroscope; 204-a second condenser lens; 205-a second photodetector; 206-dichroic mirror; 207-a collecting lens; 208-a first condenser lens; 209-a first photodetector; 3-an optical instant detector; 301-a light source emitter; 302-a fiber coupling lens; 305-a Y-shaped fiber bundle; 303-a focusing lens; 304-a fluorescence intensity detector; 306-an optical fiber entrance section; 307-fiber exit segment; 308-common segment; 309-a mirror; 310-an entrance end face; 312-an exit end face; 311-a fluorescence-receiving end face; 2-optical system of the optical sensor.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. 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.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
In the description of the present invention, it should be noted that if the terms "upper", "lower", "inside", "outside", etc. indicate an orientation or a positional relationship based on that shown in the drawings or that the product of the present invention is used as it is, this is only for convenience of description and simplification of the description, and it does not indicate or imply that the device or the element referred to must have a specific orientation, be constructed in a specific orientation, and be operated, and thus should not be construed as limiting the present invention.
Furthermore, the appearances of the terms "first," "second," and the like, if any, are used solely to distinguish one from another and are not to be construed as indicating or implying relative importance.
It should be noted that the features of the embodiments of the present invention may be combined with each other without conflict.
Referring to fig. 2, the present embodiment provides an optical real-time detector 3, including:
a light source emitter 301, a Y-shaped fiber bundle 305, a fluorescence intensity detector 304, and a mirror 309.
The Y-shaped optical fiber bundle 305 has an incident optical fiber section having an incident end face 310, a light source emitter 301 facing the incident end face 310, an emergent optical fiber section having an emergent end face 312, the emergent end face 312 facing the fluorescence intensity detector 304, and a common section 308 connected to the incident optical fiber section and the emergent optical fiber section at the same time, wherein the common section 308 has a fluorescence receiving end face 311 for transmitting light emitted from the light source emitter 301 and receiving fluorescence. In the light intensity calibration, the reflecting mirror 309 is disposed opposite to the fluorescence-receiving end surface 311.
Preferably, a fiber coupling lens 302 for coupling the emitted light into the Y-shaped fiber bundle 305 is disposed between the light source emitter 301 and the Y-shaped fiber bundle 305.
If the fiber coupling lens 302 is not provided, in order to make the light beam be better incident into the fiber, there are higher requirements on the incident angle, the number and thickness of the fiber bundle, etc. The fiber coupling lens 302 is arranged to facilitate the light emitted by the light source emitter 301 to be gathered and coupled into the Y-shaped fiber bundle 305, thereby reducing the assembly difficulty and the part requirement of the detector.
Preferably, a focusing lens 303 for focusing the emergent fluorescence to be captured by the fluorescence intensity detector 304 is disposed between the Y-shaped fiber bundle 305 and the fluorescence intensity detector 304.
Similar to the purpose of the fiber coupling lens 302, the focusing lens 303 is provided to collect the fluorescence emitted from the emitting fiber segment through the focusing lens 303 and capture the fluorescence by the fluorescence intensity detector 304, so as to obtain accurate fluorescence intensity data.
In the present embodiment, specifically, the light source emitter 301, the fiber coupling lens 302, the incident end face 310, the exit end face 312, the focusing lens 303, and the fluorescence intensity detector 304 are coaxially disposed.
Preferably, the optical instant detector 3 further comprises a sample container 1 disposed opposite to the fluorescence-receiving end face 311 in use, the sample container 1 comprising a bottom plate 11 and a cover plate 12 having light transmittance.
The sample container 1 is used for containing a sample 101 to be tested, during testing, the sample 101 to be tested is placed on the bottom plate 11, the cover plate 12 is covered, and the sample container 1 is placed on the objective table 4, so that the fluorescence receiving end surface 311 faces downwards and faces the sample 101 to be tested. In the light intensity calibration, the mirror 309 is placed on the stage 4 so that the fluorescence-receiving end surface 311 faces downward toward the mirror 309.
The present embodiment provides two working states of the optical instantaneous detector 3: a light intensity calibration state and a test state. The operating position 401 is the position where the fluorescence-receiving end surface 311 faces downward.
And (3) light intensity calibration state: when the mirror 309 is in the working position 401 on the stage 4, the detection system is in a light intensity calibration state, as shown in fig. 3. At this time, the illumination light emitted from the light source emitter 301 passes through the fiber incident section 306 and then irradiates the reflecting surface of the reflector 309, and the reflector 309 reflects a part of the illumination light to the fiber exit section 307 of the Y-shaped fiber bundle 305, and the illumination light is collected on the fluorescence intensity detector 304 through the focusing lens 303. The reference light intensity detected by the photosensor at this time is proportional to the illumination light in the case of system stabilization: i is ref =kI in （k<1）。I ref Detecting the reference light intensity on the fluorescence intensity sensor; i is in Is the emitted light intensity of the light source emitter 301.
When the sample 101 to be measured is located at the working position 401, as shown in fig. 4, the optical instantaneous detector 3 is in a measurement state. At this time, the illumination light emitted by the light source emitter 301 passes through the fiber incident section 306 of the Y-shaped fiber bundle 305, and then exits through the exit end face 312 to be irradiated onto the sample 101 to be measured, wherein the sample 101 to be measured contains a fluorescent substance, and after receiving the illumination light, fluorescence is excited. The fluorescence enters the fiber exit segment 307 of the Y-shaped fiber bundle 305 through the fluorescence receiving end face 311 and exits therefromThe end surface 312 is collected by the focusing lens 303 on the fluorescence intensity detector 304 after exiting. The fluorescence intensity detected by the fluorescence intensity detector 304 is I fl ，I fl Proportional to the content or concentration of the substance to be measured:
。
during the actual measurement, the light I is irradiated in There may be some fluctuation due to I in Is caused by a deviation of fl Deviation occurs, which causes deviation of Ctarget measurement result, so that the illumination intensity I is required to be measured on the sample in And (6) carrying out calibration. This application takes place to go on before this sample measurement to the process of light intensity calibration, because the change of light intensity is a slowly changing process, and the time interval between light intensity calibration state and the measurement state is very little, and it is suitable to set up like this. When the optical sensor is in the light intensity calibration state, the mirror 309 is placed in the working position 401, and the reference light intensity detected by the fluorescence intensity detector 304 is
. When the irradiation light intensity is the initial light intensity
When the intensity of the reference light is
. When the irradiation light is deviated, the reference light intensity is measured every time
If, if
Then the actual illumination intensity
Where the actual light intensity isConcentration of the analyte obtained under irradiation
Converted into the concentration of the original light intensity
。
Based on the above device and principle, the present application also provides a sample instant detection method, comprising: the optical instantaneous detector 3 provided by the embodiment is used for detecting the sample to obtain the actual concentration of the sample.
The method specifically comprises the following steps:
s1, determining a standard curve
Under the condition that the system is stable, the optical instantaneous detector 3 is adopted to detect a plurality of standard samples with known concentrations to obtain a plurality of fluorescence intensity values, and a standard curve of the fluorescence intensity and the concentration of the sample 101 to be detected is determined according to the plurality of known concentrations and the corresponding plurality of fluorescence intensity values
。
S2, calibrating light intensity of device
i. Turning on the light source 201, and turning on the light source 201 after receiving the detection instruction;
after the light source 201 is turned on, the mirror 309 is placed in the working position 401;
after the mirror 309 is placed in the working position 401, the fluorescence intensity detector 304 detects the reference light intensity
;
Calculating the light intensity calibration coefficient
，
A set value for the light source emitter 301 going out of field;
s3, sample detection
v. move mirror 309 out of operating position 401;
placing the sample 101 to be tested in a working position 401;
after the sample 101 to be measured is placed at the working position 401, the fluorescent substance in the sample is irradiated by the light source 201 to be excited and emit fluorescence, and the fluorescence intensity detector 304 measures the fluorescence intensity value
；
Combining the fluorescence intensity values with the standard curve
Calculating the concentration of the target substance;
calibrating the measured concentration to the actual concentration according to the light intensity calibration coefficient obtained in the step iv:
；
outputting the actual concentration value;
xi, the sample to be measured 101 moves out of the working position 401, and the reflector 309 enters the working position 401 when the next measurement is waited;
in performing steps v to xi, the optical instantaneous detector 3 is in a sample detection state.
The sample instant detection method provided by the application is very suitable for being applied to medical examination, and can be used for conveniently and rapidly detecting the biological sample of a patient.
In summary, the optical instant detector provided by the present application can perform optical path calibration by disposing the reflecting mirror at a position opposite to the fluorescence receiving end face due to a reasonable structural arrangement, and does not need to separately dispose an optical path as in the conventional detector mentioned in the background art. The detector has a simple structure, parts are obviously less than those of the existing detectors, and the detector can be integrated into equipment which has a small volume and is convenient to take and use; because of the few optical parts, the manufacturing cost is lower.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (8)
1. An optical instantaneous detector, characterized in that it comprises:
the device comprises a light source emitter, a Y-shaped optical fiber bundle, a fluorescence intensity detector and a reflector;
the Y-shaped optical fiber bundle is provided with an incident optical fiber section, an emergent optical fiber section and a common section which is simultaneously connected with the incident optical fiber section and the emergent optical fiber section, the incident optical fiber section is provided with an incident end face, the light source emitter faces the incident end face, the emergent optical fiber section is provided with an emergent end face, the emergent end face faces the fluorescence intensity detector, and the common section is provided with a fluorescence receiving end face which is used for transmitting light rays emitted by the light source emitter and can receive fluorescence;
and when the light intensity is calibrated, the reflecting mirror is arranged opposite to the fluorescence receiving end face.
2. The optical instant detector of claim 1 wherein a fiber coupling lens is disposed between the light source emitter and the Y-shaped fiber bundle for coupling emitted light into the Y-shaped fiber bundle.
3. The optical instant detector of claim 1 wherein a focusing lens is disposed between the Y-shaped fiber bundle and the fluorescence intensity detector for focusing the exiting fluorescence to be captured by the fluorescence intensity detector.
4. The optical instant detector of claim 1 further comprising a sample vessel disposed in use opposite the fluorescence-receiving end face, the sample vessel comprising a bottom plate and a cover plate having optical transparency.
5. A method for real-time sample detection, which is characterized in that the optical real-time detector according to any one of claims 1 to 4 is used for sample detection, and comprises the following steps:
light intensity calibration: the reflector is arranged at the position opposite to the fluorescence receiving end surface for light intensity calibration, and the fluorescence intensity detector is used for measuring the reference light intensity
According to the formula
The light intensity calibration factor is calculated,
setting a light intensity value for the factory of the light source emitter, wherein alpha is a light intensity calibration coefficient;
sample detection: and (3) placing the sample at a position opposite to the fluorescence receiving end surface to detect the fluorescence intensity of the sample, and calculating to obtain the actual concentration of the sample according to the measured fluorescence intensity, the light intensity calibration coefficient and the standard curve relation between the fluorescence intensity and the concentration of the sample to be detected.
6. The method according to claim 5, wherein the sample testing step comprises:
placing a sample to be detected at a position opposite to the fluorescence receiving end face, and measuring the fluorescence intensity of a fluorescent substance in the sample to be detected, wherein the fluorescent substance is irradiated by a light source and then stimulated to emit the fluorescence intensity through the fluorescence intensity detector
Calculating to obtain the test concentration of the target object according to the standard curve
According to the formula
Obtaining the actual concentration
。
7. The method for on-line testing of samples according to claim 5, further comprising, prior to testing: and detecting a plurality of standard samples with known concentrations by using the optical instant detector to obtain a plurality of fluorescence intensity values, and determining a standard curve of the fluorescence intensity and the concentration of the sample to be detected according to the plurality of known concentrations and the plurality of corresponding fluorescence intensity values.
8. Use of the method of any one of claims 5 to 7 for the point-of-care testing of a sample in medical testing.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117348224A (en) * | 2023-12-04 | 2024-01-05 | 成都丹诺迪医疗科技有限公司 | Microscopic device |
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Denomination of invention: Optical instant detector and detection method and application Effective date of registration: 20220831 Granted publication date: 20220208 Pledgee: Bank of Chengdu science and technology branch of Limited by Share Ltd. Pledgor: Sichuan danuodi Technology Co.,Ltd. Registration number: Y2022510000258 |