CN115015204B - Fluorescence detection device - Google Patents
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- CN115015204B CN115015204B CN202210718047.6A CN202210718047A CN115015204B CN 115015204 B CN115015204 B CN 115015204B CN 202210718047 A CN202210718047 A CN 202210718047A CN 115015204 B CN115015204 B CN 115015204B
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- 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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- 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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- 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
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
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- 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
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- 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
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Abstract
The invention relates to the technical field of fluorescence detection, in particular to a fluorescence detection device, which comprises an excitation light source part, a sample cell part and a fluorescence detection part, wherein the excitation light source part comprises a light source and an excitation light channel; the fluorescence detection part comprises a detector and a fluorescence channel; the exciting light channel comprises a main exciting light channel and an auxiliary exciting light channel, the fluorescence channel comprises a main fluorescence channel and an auxiliary fluorescence channel, and the main exciting light channel and the main fluorescence channel are both connected with the sample cell part; the auxiliary exciting light channel is communicated with the main exciting light channel, and the auxiliary fluorescence channel is communicated with the main fluorescence channel, so that exciting light in the auxiliary exciting light channel irradiates the sample cell part through the main exciting light channel, a sample in the sample cell part is excited to emit fluorescence, and the fluorescence is irradiated to the detector of the corresponding main fluorescence channel or the detector of the auxiliary fluorescence channel through the main fluorescence channel; one detector corresponds to at least one light source. The invention has the advantages of channel multiplexing and detector multiplexing, thereby improving the luminous flux and saving the cost.
Description
Technical Field
The invention relates to the technical field of fluorescence detection, in particular to a fluorescence detection device which is mainly used for realizing channel multiplexing and detector multiplexing, reducing cost and improving luminous flux.
Background
Fluorescence has been widely used in biological, chemical, medical and other fields, and fluorescence detection devices achieve related analysis by detecting fluorescence, wherein the main principle of the fluorescence detection device is that a light source emits excitation light to irradiate a sample, a fluorescent substance in the sample emits fluorescence after being induced by the excitation light, and a detector achieves corresponding analysis by detecting the fluorescence. Exciting light emitted by the light source passes through a plurality of optical filters on the way of irradiating the sample, and the thinking of separation, collimation and convergence is adopted; the fluorescence generated after the excitation light excites the sample and needs to pass through a plurality of filters to irradiate the detector.
Most of the prior art are detectors for detecting one fluorescence, and each excitation light can only induce one fluorescence, so that a large number of optical filters are needed when detecting various fluorescences, and the detectors with the same number as the light sources are needed, so that the cost is high; the passage of the excitation light is also relatively narrow in a limited space, resulting in a limited light passage.
Disclosure of Invention
The invention provides a fluorescence detection device, which adopts channel multiplexing and detector multiplexing, reduces the number of optical filters and detectors, saves cost, and improves luminous flux due to channel multiplexing.
The invention provides a fluorescence detection device, which comprises an excitation light source part, a sample cell part and a fluorescence detection part, wherein the excitation light source part comprises a light source and an excitation light channel matched with the light source; the fluorescence detection part comprises a detector and a fluorescence channel matched with the detector;
the excitation light channel comprises a main excitation light channel and an auxiliary excitation light channel, the fluorescence channel comprises a main fluorescence channel and an auxiliary fluorescence channel, and the main excitation light channel and the main fluorescence channel are both connected with the sample cell part;
the auxiliary excitation light channel is communicated with the main excitation light channel, and the auxiliary fluorescence channel is communicated with the main fluorescence channel, so that the excitation light in the auxiliary excitation light channel is irradiated to the sample cell part through the main excitation light channel, a sample in the sample cell part emits fluorescence after being excited by the excitation light, and the fluorescence is irradiated to the detector of the corresponding main fluorescence channel or the detector of the auxiliary fluorescence channel through the main fluorescence channel;
one of the detectors is used for correspondingly detecting fluorescence emitted by exciting the sample in the sample cell part by the exciting light emitted by at least one of the light sources.
Optionally, a dichroic mirror is arranged on the main excitation light channel, the dichroic mirror is arranged at a connection position of the main excitation light channel and the auxiliary excitation light channel, and the excitation light on the auxiliary excitation light channel is reflected onto the main excitation light channel through the dichroic mirror.
Optionally, the secondary excitation light channel is vertically connected to the main excitation light channel, and an included angle between the dichroic mirror and the secondary excitation light channel is 1/2 of an included angle between the secondary excitation light channel and the main excitation light channel.
Optionally, the secondary excitation light channel is provided in plurality, and the number of the dichroic mirrors on the primary excitation light channel is the same as the number of the secondary excitation light channels.
Optionally, a first optical filter is disposed on the main fluorescence channel, and the first optical filter is disposed at a joint of the main fluorescence channel and the auxiliary fluorescence channel, so that fluorescence corresponding to the auxiliary fluorescence channel is reflected to the auxiliary fluorescence channel through the first optical filter.
Optionally, the secondary fluorescent channel is vertically connected with the primary fluorescent channel; the included angle between the first optical filter and the auxiliary fluorescent channel is 1/2 of the included angle between the auxiliary fluorescent channel and the main fluorescent channel.
Optionally, a narrow-band filter is disposed on the excitation light channel at a position close to the light source, and the narrow-band filter on each excitation light channel has different specifications.
Optionally, a notch filter is disposed on the fluorescence channel at a position close to the detector.
Optionally, the excitation light channel is provided with an aspheric lens at a position close to the light source and at a position close to the sample cell part; and aspheric lenses are arranged at the positions, close to the detector, and the sample cell part on the fluorescence channel.
Optionally, the excitation light source part, the sample cell part and the fluorescence detection part are integrally formed.
Advantageous effects
The invention provides a fluorescence detection device, which comprises an excitation light source part, a sample cell part and a fluorescence detection part, wherein the excitation light source part comprises a light source and an excitation light channel matched with the light source; the fluorescence detection part comprises a detector and a fluorescence channel matched with the detector; the excitation light channel comprises a main excitation light channel and an auxiliary excitation light channel, the fluorescence channel comprises a main fluorescence channel and an auxiliary fluorescence channel, and the main excitation light channel and the main fluorescence channel are both connected with the sample cell part; exciting light emitted by the light source is irradiated to the sample cell part through the main exciting light channel, and a sample in the sample cell part is excited by the exciting light to emit fluorescence to be irradiated to the main fluorescence channel; the auxiliary excitation light channel is communicated with the main excitation light channel, and the auxiliary fluorescence channel is communicated with the main fluorescence channel, so that the excitation light in the auxiliary excitation light channel is irradiated to the sample cell part through the main excitation light channel, a sample in the sample cell part emits fluorescence after being excited by the excitation light, and the fluorescence is irradiated to the detector of the corresponding main fluorescence channel or the detector of the auxiliary fluorescence channel through the main fluorescence channel; further realize that: no matter which light source emits the exciting light, the exciting light can be emitted to the sample cell part through the main exciting light channel, and no matter which exciting light excites the fluorescence, the exciting light is firstly emitted into the main fluorescence channel and then is emitted into the corresponding main fluorescence channel or the auxiliary fluorescence channel according to the specific detection requirement; in addition, one of the detectors correspondingly detects fluorescence emitted by exciting the sample in the sample cell portion by the excitation light emitted by at least one of the light sources, wherein if one of the detectors corresponds to a plurality of the light sources, the plurality of the light sources emit the excitation light at different times; the detector can detect multiple kinds of fluorescence by realizing time-sharing multiplexing, namely that one detector can detect multiple kinds of excitation light, but when the detector is used, the light source emits the excitation light at different times to realize that the corresponding fluorescence arrives at the detector at different times. The invention has the advantages of multiplexing the main exciting light channel and the main fluorescence channel and multiplexing the detector, improving the luminous flux and saving the cost.
The invention also has the following beneficial effects:
1. the dichroic mirror is arranged on the main excitation light channel and is arranged at the joint of the main excitation light channel and the auxiliary excitation light channel, so that excitation light in the main excitation light channel penetrates through the dichroic mirror to irradiate the sample pool, and the excitation light in the auxiliary excitation light channel is reflected into the main excitation light channel by the dichroic mirror and then enters the sample pool along the main excitation light channel, so that channel multiplexing is realized.
2. The auxiliary exciting light channel is vertically connected with the main exciting light channel, so that the arrangement design is convenient, and the space is effectively utilized; the contained angle of dichroic mirror and vice exciting light passageway is 1/2 of vice exciting light passageway and main exciting light passageway contained angle, is adapted to the design of vice exciting light passageway to realize that the exciting light in the vice exciting light passageway can accurately reflect into main exciting light passageway.
3. The auxiliary exciting light channels are provided with a plurality of fluorescent lights, more fluorescent lights can be conveniently detected, the detection efficiency is improved, the number of the dichroic mirrors on the main exciting light channel is consistent with that of the auxiliary exciting light channels, and the exciting light on each auxiliary exciting light channel can be reflected into the main exciting light channel through the corresponding dichroic mirrors.
4. The narrow-band filters are arranged on the excitation light channels and close to the light source, the narrow-band filters on each excitation light channel are different in specification and used for allowing excitation light with specific wavelength to pass through and filtering the excitation light emitted by the light source, so that the specific excitation light is used for exciting the specific fluorescent substance to emit corresponding fluorescent light, and the detection and analysis of the specific fluorescent substance in the sample are facilitated.
5. The aspheric lens can better collimate the exciting light to reach the sample cell part, and can better collect the fluorescence to the corresponding detector, thereby being convenient for improving the detection precision.
6. The excitation light source part, the sample cell part and the fluorescence detection part are integrally formed, so that the consistency of spatial positions is ensured, the accuracy of an angle between the incident excitation light and the emergent fluorescence of the sample cell part is ensured, and the stability of fluorescence detection is ensured.
It should be understood that the statements herein reciting aspects are not intended to limit the critical or essential features of any embodiment of the invention, nor are they intended to limit the scope of the invention. Other features of the present invention will become apparent from the following description.
Drawings
The above and other features, advantages and aspects of various embodiments of the present invention will become more apparent by referring to the following detailed description when taken in conjunction with the accompanying drawings. In the drawings, the same or similar reference numerals denote the same or similar elements.
FIG. 1 is a schematic diagram showing a fluorescence detection device according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of the light path of a fluorescence detection device according to an embodiment of the present invention;
FIG. 3 shows a schematic diagram of components of a fluorescence detection apparatus according to an embodiment of the present invention.
Wherein, 1-excitation light source part; 1-1-a light source; 1-1 a-a first light source; 1-1 b-a second light source; 1-1 c-a third light source; 1-1 d-a fourth light source; 1-2-main excitation light channel; 1-21-dichroic mirror; 1-22-narrow band filters; 1-23-aspheric lens; 1-3-secondary excitation light channels; 2-a sample cell portion; 3-a fluorescent detection moiety; 3-1-a detector; 3-1 a-a first detector; 3-1 b-a second detector; 3-1 c-a third detector; 3-2-primary fluorescence channel; 3-21-a first filter; 3-22-a notch filter; 3-3-secondary fluorescence channel.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in one or more embodiments of the present specification, the technical solutions in one or more embodiments of the present specification will be clearly and completely described below with reference to the drawings in one or more embodiments of the present specification, and it is obvious that the described embodiments are only a part of the embodiments of the present specification, and not all embodiments. All other embodiments that can be derived by a person skilled in the art from one or more of the embodiments described herein without making any inventive step shall fall within the scope of protection of this document.
It should be noted that, the description in the embodiment of the present invention is only for clearly illustrating the technical solutions in the embodiment of the present invention, and does not limit the technical solutions provided in the embodiment of the present invention.
FIG. 1 is a schematic diagram showing a fluorescence detection device according to an embodiment of the present invention; FIG. 2 is a schematic diagram of the light path of a fluorescence detection device according to an embodiment of the present invention; referring to fig. 1 and 2, the fluorescence detection apparatus includes an excitation light source portion 1, a sample cell portion 2, and a fluorescence detection portion 3, where the excitation light source portion 1 includes a light source 1-1 and an excitation light channel matched with the light source 1-1; the fluorescence detection part 3 comprises a detector 3-1 and a fluorescence channel matched with the detector 3-1; the excitation light channel comprises a main excitation light channel 1-2 and an auxiliary excitation light channel 1-3, the fluorescence channel comprises a main fluorescence channel 3-2 and an auxiliary fluorescence channel 3-3, and the main excitation light channel 1-2 and the main fluorescence channel 3-2 are both connected with the sample cell part 2; the secondary excitation light channel 1-3 is communicated with the primary excitation light channel 1-2, and the secondary fluorescence channel 3-3 is communicated with the primary fluorescence channel 3-2, so that the excitation light in the secondary excitation light channel 1-3 is irradiated to the sample cell part 2 through the primary excitation light channel 1-2, the sample in the sample cell part 2 is excited by the excitation light to emit fluorescence, and the fluorescence is irradiated to the detector 3-1 of the corresponding primary fluorescence channel 3-2 or the detector 3-1 of the secondary fluorescence channel 3-3 through the primary fluorescence channel 3-2; one of the detectors 3-1 correspondingly detects fluorescence emitted by excitation light emitted from at least one of the light sources 1-1 exciting the sample in the sample cell portion 2.
In this embodiment, a light source 1-1 is configured to emit excitation light, each light source 1-1 corresponds to an excitation light channel, the excitation light channels in this embodiment include a main excitation light channel 1-2 and an auxiliary excitation light channel 1-3, and the main excitation light channel 1-2 and the auxiliary excitation light channel 1-3 both correspond to respective light sources 1-1, wherein the main excitation light channel 1-2 is connected to a sample cell portion 2, excitation light emitted by the light source 1-1 corresponding to the main excitation light channel 1-2 can directly enter the sample cell portion 2, while excitation light emitted by the light source 1-1 corresponding to the auxiliary excitation light channel 1-3 first enters the main excitation light channel 1-2 and then enters the sample cell portion 2 along the main excitation light channel 1-2, that is, regardless of the excitation light emitted by the main excitation light channel 1-2 or the excitation light emitted by the auxiliary excitation light channel 1-3, angles of all the excitation light to the sample cell portion 2 are the same; correspondingly, the fluorescence channel in this embodiment includes a main fluorescence channel 3-2 and a secondary fluorescence channel 3-3, the main fluorescence channel 3-2 and the secondary fluorescence channel 3-3 are respectively corresponding to the respective detectors 3-1, wherein the main fluorescence channel 3-2 is connected to the sample cell portion 2, the corresponding fluorescent substance in the sample of the sample cell portion 2 is excited by the incident excitation light to emit fluorescence, the detector 3-1 is used for detecting the fluorescence emitted by the excitation light emitted by the corresponding light source 1-1 to excite the fluorescent substance, and during detection, no matter which excitation light emitted by the corresponding light source 1-1 excites the fluorescence emitted by the corresponding fluorescent substance, the fluorescence is firstly emitted into the main fluorescence channel 3-2, and then emitted into the corresponding main fluorescence channel 3-2 or secondary fluorescence channel 3-3 according to the difference of the corresponding fluorescence wavelength, and detection is performed by the corresponding detector 3-1. In this embodiment, one detector 3-1 can correspondingly detect fluorescence corresponding to excitation light emitted by a plurality of different light sources 1-1, that is, the detector 3-1 and the light source 1-1 may be in a one-to-one relationship or a one-to-many relationship, and the number of the detectors 3-1 is less than the number of the light sources 1-1.
The embodiment provides a fluorescence detection device, which comprises an excitation light source part 1, a sample cell part 2 and a fluorescence detection part 3, wherein the excitation light source part 1 comprises a light source 1-1 and an excitation light channel matched with the light source 1-1; the fluorescence detection part 3 comprises a detector 3-1 and a fluorescence channel matched with the detector 3-1; the exciting light channel comprises a main exciting light channel 1-2 and an auxiliary exciting light channel 1-3, the fluorescence channel comprises a main fluorescence channel 3-2 and an auxiliary fluorescence channel 3-3, and the main exciting light channel 1-2 and the main fluorescence channel 3-2 are both connected with the sample cell part 2; the secondary excitation light channel 1-3 is communicated with the primary excitation light channel 1-2, and the secondary fluorescence channel 3-3 is communicated with the primary fluorescence channel 3-2, so that the excitation light in the secondary excitation light channel 1-3 is irradiated to the sample cell part 2 through the primary excitation light channel 1-2, the sample in the sample cell part 2 is excited by the excitation light to emit fluorescence, and the fluorescence is irradiated to the detector 3-1 of the corresponding primary fluorescence channel 3-2 or the detector 3-1 of the secondary fluorescence channel 3-3 through the primary fluorescence channel 3-2; one of the detectors 3-1 correspondingly detects fluorescence emitted by exciting the sample in the sample cell portion 2 by at least one of the light sources 1-1; the time-sharing multiplexing of the detector 3-1 can be realized, namely, when one detector 3-1 can detect multiple fluorescence correspondingly excited by multiple excitation lights, when in use, the light source 1-1 emits the excitation lights at different times to realize that the corresponding fluorescence arrives at the detector 3-1 at different times, and the detector 3-1 can detect the multiple fluorescence. The main exciting light channel 1-2 and the main fluorescence channel 3-2 are multiplexed, and the detector 3-1 is multiplexed, so that the luminous flux is improved, and the cost is saved.
Specifically, dichroic mirrors 1 to 21 are arranged on the main excitation light channels 1 to 2, the dichroic mirrors 1 to 21 are arranged at the connection positions of the main excitation light channels 1 to 2 and the auxiliary excitation light channels 1 to 3, and the excitation light on the auxiliary excitation light channels 1 to 3 is reflected to the main excitation light channels 1 to 2 through the dichroic mirrors 1 to 21. The dichroic mirror 1-21 can transmit excitation light on the main excitation light channel 1-2, and reflect excitation light on the auxiliary excitation light channel 1-3 to the main excitation light channel 1-2, wherein the "excitation light on the main excitation light channel 1-2" includes excitation light emitted by the light source 1-1 corresponding to the main excitation light channel 1-2, and excitation light emitted by the light source 1-1 corresponding to the other auxiliary excitation light channels 1-3 and reflected into the main excitation light channel 1-2. In this embodiment, the auxiliary excitation light channel 1-3 is vertically connected to the main excitation light channel 1-2; the included angle between the dichroic mirror 1-21 and the secondary excitation light channel 1-3 is 1/2 of the included angle between the secondary excitation light channel 1-3 and the main excitation light channel 1-2. The auxiliary excitation light channel 1-3 is vertically connected with the main excitation light channel 1-2, so that the layout of the light source 1-1 and the layout of the dichroic mirror 1-21 are facilitated, when viewed in the clockwise direction, if the auxiliary excitation light channel 1-3 is arranged on the left side of the main excitation light channel 1-2 (when viewed in the excitation light irradiation direction, refer to fig. 1), the included angle between the main excitation light channel 1-2 and the auxiliary excitation light channel 1-3 is 270 degrees, and the included angle between the dichroic mirror 1-21 and the auxiliary excitation light channel 1-3 is 135 degrees; if the auxiliary excitation light channel 1-3 is arranged on the right side of the main excitation light channel 1-2, the included angle between the main excitation light channel 1-2 and the auxiliary excitation light channel 1-3 is 90 degrees, and the included angle between the dichroic mirror 1-21 and the auxiliary excitation light channel 1-3 is 45 degrees. In this embodiment, there are a plurality of secondary excitation light channels 1-3, and the number of dichroic mirrors 1-21 on the primary excitation light channel 1-2 is the same as the number of secondary excitation light channels 1-3. That is, each secondary excitation light channel 1-3 corresponds to one dichroic mirror 1-21 on the primary excitation light channel 1-2, the dichroic mirrors 1-21 are arranged according to the arrangement of the corresponding secondary excitation light channels 1-3, and the secondary excitation light channels 1-3 can be arranged on the left side or the right side of the primary excitation light channel 1-2 according to different specific spaces. In this embodiment, only one of the sub-excitation light channels 1 to 3 may be provided.
Specifically, a first optical filter 3-21 is disposed on the main fluorescent channel 3-2, and the first optical filter 3-21 is disposed at a joint of the main fluorescent channel 3-2 and the auxiliary fluorescent channel 3-3, so that the fluorescence corresponding to the auxiliary fluorescent channel 3-3 is reflected to the auxiliary fluorescent channel 3-3 through the first optical filter 3-21. And the fluorescence corresponding to the main fluorescence channel 3-2 and the fluorescence corresponding to the other auxiliary fluorescence channel 3-3 are transmitted through the first filter 3-21. The specification of the first filter 3-21 can be different types of filters or different specifications of filters according to the specific required transmission or reflection of fluorescence. In this embodiment, the secondary fluorescent channel 3-3 is vertically connected to the primary fluorescent channel 3-2; the included angle between the first optical filter 3-21 and the auxiliary fluorescent channel 3-3 is 1/2 of the included angle between the auxiliary fluorescent channel 3-3 and the main fluorescent channel 3-2. The auxiliary fluorescent channel 3-3 is vertically connected with the main fluorescent channel 3-2, so that the layout of the detector 3-1 and the layout of the first optical filter 3-21 are facilitated, when viewed in a clockwise direction, if the auxiliary fluorescent channel 3-3 is arranged on the left side of the main fluorescent channel 3-2 (when viewed in a fluorescent irradiation direction, refer to fig. 1), the included angle between the main fluorescent channel 3-2 and the auxiliary fluorescent channel 3-3 is 90 degrees, and the included angle between the first optical filter 3-21 and the auxiliary fluorescent channel 3-3 is 45 degrees; if the secondary fluorescent channel 3-3 is arranged on the right side of the primary fluorescent channel 3-2, the included angle between the primary fluorescent channel 3-2 and the secondary fluorescent channel 3-3 is 270 degrees, and the included angle between the first optical filter 3-21 and the secondary fluorescent channel 3-3 is 135 degrees.
Specifically, narrow-band filters 1-22 are arranged on the excitation light channels at positions close to the light source 1-1, and the specifications of the narrow-band filters 1-22 on each excitation light channel are different. The exciting light emitted by the light source 1-1 is firstly filtered by the narrow band filters 1-22, so that the exciting light with a specific wavelength can penetrate through the narrow band filters to realize that the specific fluorescent substance is singly excited to emit fluorescence, the wavelengths of the penetrating exciting light after the exciting light emitted by the light sources 1-1 is filtered by the corresponding narrow band filters 1-22 are different, the multiple fluorescent substances in a sample can be detected in a one-to-one correspondence manner instead of mixed detection, the detection and analysis of the specific fluorescent substance are facilitated, the influence of other substances on the detection result is excluded, and the detection accuracy is improved.
Specifically, a notch filter 3-22 is arranged on the fluorescence channel at a position close to the detector 3-1. The notch filters 3 to 22 attenuate fluorescence of a specific wavelength band and reduce scattering of light.
Specifically, aspheric lenses 1-23 are arranged on the excitation light channel at positions close to the light source 1-1 and the sample cell part 2; and aspheric lenses 1-23 are arranged on the fluorescence channel at positions close to the detector 3-1 and the sample cell part 2. The lens mainly functions to converge light to one point, in the embodiment, the aspheric lenses 1-23 are adopted and used for focusing and collimating the exciting light emitted by the light source 1-1 and then sending the light to the sample cell part 2, and the aspheric lenses 1-23 are arranged at positions close to the sample cell part 2 and can further focus and collimate the light, so that the exciting light can be more irradiated to the sample cell part 2, and the detection precision is improved; the fluorescence emitted from the sample cell part 2 is focused and collimated by the aspheric lenses 1-23 and then sent to the corresponding fluorescence channel, and before reaching the detector 3-1, the fluorescence is further focused and collimated by the aspheric lenses 1-23, so that the fluorescence is more detected by the detector 3-1. The aspheric lenses 1-23 can increase the effective focal length and eliminate spherical aberration to the maximum extent, so that the exciting light can be better focused on the sample cell part 2, and the fluorescence can be better focused on the detector 3-1.
Specifically, the excitation light source portion 1, the sample cell portion 2, and the fluorescence detection portion 3 are integrally formed. For example, the fluorescence detection device can be formed by precision injection molding, the errors caused by processing and assembly can be reduced to the smallest possible extent, the consistency of the spatial positions of the excitation light source part 1, the sample cell part 2 and the fluorescence detection part 3 can be ensured, and the stability of fluorescence detection can be further ensured.
The advantageous effects of the invention are illustrated below in a preferred embodiment:
FIG. 3 shows a schematic component view of a fluorescence detection device according to an embodiment of the present invention; referring to fig. 3, the excitation light source portion 1 includes four light sources 1-1 and four excitation light channels corresponding to the four light sources 1-1, where the four light sources 1-1 are a first light source 1-1a, a second light source 1-1b, a third light source 1-1c, and a fourth light source 1-1d, respectively, the excitation light channel corresponding to the first light source 1-1a is a main excitation light channel 1-2, and the second light source 1-1b, the third light source 1-1c, and the fourth light source 1-1d correspond to four sub-excitation light channels 1-3, respectively. The fluorescence detection part 3 comprises three detectors 3-1 and three fluorescence channels corresponding to the three detectors 3-1, wherein the three detectors 3-1 are respectively a first detector 3-1a, a second detector 3-1b and a third detector 3-1c, the fluorescence channel corresponding to the first detector 3-1a is a main fluorescence channel 3-2, and the fluorescence channel corresponding to the second detector 3-1b and the third detector 3-1c is a secondary fluorescence channel 3-3. Wherein, the first detector 3-1a correspondingly detects the fluorescence emitted by the corresponding fluorescent substance after the excitation light emitted by the third light source 1-1c excites the corresponding fluorescent substance; the second detector 3-1b correspondingly detects the fluorescence emitted by the corresponding fluorescent substance after the excitation light emitted by the second light source 1-1b excites the corresponding fluorescent substance; the third detector 3-1c correspondingly detects the fluorescence emitted by the excitation light emitted from the first light source 1-1a and the fourth light source 1-1d after exciting the corresponding fluorescent substance.
The exciting light emitted by the first light source 1-1a passes through the aspheric lens 1-23, the exciting light emitted by the first light source 1-1a is firstly focused and collimated, and then is irradiated onto the corresponding narrowband filter 1-22, the narrowband filter 1-22 screens the exciting light, only the exciting light with a specific wavelength is allowed to pass through, the exciting light which passes through the narrowband filter 1-22 is directly irradiated to the sample cell part 2, wherein before being irradiated to the sample cell part 2, the exciting light is firstly focused and collimated through the aspheric lens 1-23, and the exciting light irradiated to the sample cell part 2 excites the corresponding fluorescent substance in the sample to emit fluorescence. The fluorescence firstly passes through the notch filters 3-22, and the notch filters 3-22 can avoid the transmission of the fluorescence of some wave bands which do not need to be detected, thereby improving the detection precision; the fluorescence which penetrates through the notch filter 3-22 enters the main fluorescence channel 3-2, a first filter 3-21 is arranged at the position, connected with the auxiliary fluorescence channel 3-3 corresponding to the third detector 3-1c, of the main fluorescence channel 3-2, the fluorescence is reflected to the auxiliary fluorescence channel 3-3 corresponding to the third detector 3-1c through the first filter 3-21 and is irradiated towards the direction of the third detector 3-1c along the auxiliary fluorescence channel 3-3, the notch filter 3-22 and the aspheric lens 1-23 are sequentially arranged at the position close to the third detector 3-1c, the transmittance of the fluorescence is increased through the notch filter 3-22, and then the fluorescence is focused and collimated through the aspheric lens 1-23 and is irradiated onto the third detector 3-1 c. Exciting light emitted by the second light source 1-1b, the third light source 1-1c and the fourth light source 1-1d irradiates the sample cell part 2, excites corresponding fluorescent substances to emit corresponding fluorescent light, and irradiates the corresponding detector 3-1, wherein the exciting light emitted by the second light source 1-1b irradiates the sample cell part 2, and excites the fluorescent light emitted by the corresponding fluorescent substances to be reflected to the detector 3-1b through the first optical filter 3-21; the exciting light emitted by the third light source 1-1c irradiates the sample cell part 2, and the fluorescence emitted by the corresponding fluorescent substance is excited to penetrate through the first optical filter 3-21 and be directly irradiated onto the detector 3-1 a; the excitation light from the fourth light source 1-1d is irradiated to the cuvette portion 2, and the fluorescence from the corresponding fluorescent substance is reflected to the detector 3-1c via the first filter 3-21. The detector 3-1c can correspondingly detect the fluorescence emitted by the excitation light emitted by the first light source 1-1a and the fourth light source 1-1d after exciting the corresponding fluorescent substance, and since the first light source 1-1a and the fourth light source 1-1d are turned on in a time-sharing manner, the time when the corresponding fluorescence finally reaches the detector 3-1c is different, no conflict is generated between the first light source 1-1a and the fourth light source 1-1d, and then the time-sharing multiplexing of the detector 3-1c can be realized. Any one detector 3-1 can correspondingly detect the fluorescence corresponding to the exciting light emitted by a plurality of different light sources 1-1, so that the number of the detectors 3-1 can be reduced, and the difference caused by a plurality of detectors 3-1 can be reduced.
A first optical filter 3-21 arranged at the joint of the primary fluorescence channel 3-2 and the secondary fluorescence channel 3-3 corresponding to the second detector 3-1b is a dichroic mirror; the first optical filter 3-21 arranged at the joint of the main fluorescence channel 3-2 and the auxiliary fluorescence channel 3-3 corresponding to the third detector 3-1c is a broadband pass optical filter.
The foregoing description is only exemplary of the preferred embodiments of the invention and is illustrative of the principles of the technology employed. It will be appreciated by those skilled in the art that the scope of the invention herein disclosed is not limited to the particular combination of features described above, but also encompasses other arrangements formed by any combination of the above features or their equivalents without departing from the spirit of the invention. For example, the above features and (but not limited to) features having similar functions disclosed in the present invention are mutually replaced to form the technical solution.
Claims (9)
1. The fluorescence detection device is characterized by comprising an excitation light source part (1), a sample cell part (2) and a fluorescence detection part (3), wherein the excitation light source part (1) comprises a light source (1-1) and an excitation light channel matched with the light source (1-1); the fluorescence detection part (3) comprises a detector (3-1) and a fluorescence channel matched with the detector (3-1);
the excitation light channel comprises a main excitation light channel (1-2) and an auxiliary excitation light channel (1-3), the fluorescence channel comprises a main fluorescence channel (3-2) and an auxiliary fluorescence channel (3-3), and the main excitation light channel (1-2) and the main fluorescence channel (3-2) are both connected with the sample cell part (2);
the auxiliary exciting light channel (1-3) is communicated with the main exciting light channel (1-2), the auxiliary fluorescent light channel (3-3) is communicated with the main fluorescent light channel (3-2), so that exciting light in the auxiliary exciting light channel (1-3) is irradiated to the sample cell part (2) through the main exciting light channel (1-2), a sample in the sample cell part (2) emits fluorescent light after being excited by the exciting light, and the fluorescent light is irradiated to the detector (3-1) of the corresponding main fluorescent light channel (3-2) or the detector (3-1) of the auxiliary fluorescent light channel (3-3) through the main fluorescent light channel (3-2);
one of the detectors (3-1) correspondingly detects fluorescence emitted by exciting light emitted by at least one of the light sources (1-1) to excite the sample in the sample cell portion (2);
the excitation light source part (1), the sample cell part (2) and the fluorescence detection part (3) are integrally formed.
2. A fluorescence detection device according to claim 1, wherein:
the main excitation light channel (1-2) is provided with a dichroic mirror (1-21), the dichroic mirror (1-21) is arranged at the joint of the main excitation light channel (1-2) and the auxiliary excitation light channel (1-3), and the excitation light on the auxiliary excitation light channel (1-3) is reflected to the main excitation light channel (1-2) through the dichroic mirror (1-21).
3. A fluorescence detection device according to claim 2, wherein:
the auxiliary exciting light channel (1-3) is vertically connected with the main exciting light channel (1-2); the included angle between the dichroic mirror (1-21) and the auxiliary exciting light channel (1-3) is 1/2 of the included angle between the auxiliary exciting light channel (1-3) and the main exciting light channel (1-2).
4. A fluorescence detection device according to claim 3, wherein:
the number of the secondary excitation light channels (1-3) is multiple, and the number of the dichroic mirrors (1-21) on the main excitation light channel (1-2) is consistent with that of the secondary excitation light channels (1-3).
5. A fluorescence detection device according to claim 1, wherein:
the main fluorescence channel (3-2) is provided with a first optical filter (3-21), and the first optical filter (3-21) is arranged at the joint of the main fluorescence channel (3-2) and the auxiliary fluorescence channel (3-3), so that fluorescence corresponding to the auxiliary fluorescence channel (3-3) is reflected to the auxiliary fluorescence channel (3-3) through the first optical filter (3-21).
6. A fluorescence detection device according to claim 5, wherein:
the secondary fluorescent channel (3-3) is vertically connected with the main fluorescent channel (3-2); the included angle between the first optical filter (3-21) and the auxiliary fluorescent channel (3-3) is 1/2 of the included angle between the auxiliary fluorescent channel (3-3) and the main fluorescent channel (3-2).
7. A fluorescence detection device according to claim 1, wherein:
narrow-band filters (1-22) are arranged on the excitation light channels at positions close to the light sources (1-1), and the specifications of the narrow-band filters (1-22) on each excitation light channel are different.
8. A fluorescence detection device according to claim 1, wherein:
and a notch filter (3-22) is arranged on the fluorescent channel and close to the detector (3-1).
9. A fluorescence detection device according to claim 1, wherein:
aspheric lenses (1-23) are arranged at the positions, close to the light source (1-1) and the sample cell part (2), on the exciting light channel;
and aspheric lenses (1-23) are arranged at the positions, close to the detector (3-1) and the sample cell part (2), of the fluorescence channel.
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| KR102126032B1 (en) * | 2013-07-31 | 2020-07-08 | 삼성전자주식회사 | Multi-channel fluorescence detecting module and nucleic acid analysis system having the same |
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| CN111239093A (en) * | 2020-03-13 | 2020-06-05 | 苏州雅睿生物技术有限公司 | Planar miniature multi-channel fluorescence detection optical system |
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