CN104293648B - Gene sequencing light path system - Google Patents

Gene sequencing light path system Download PDF

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Publication number
CN104293648B
CN104293648B CN201410515660.3A CN201410515660A CN104293648B CN 104293648 B CN104293648 B CN 104293648B CN 201410515660 A CN201410515660 A CN 201410515660A CN 104293648 B CN104293648 B CN 104293648B
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light
light beam
spectroscope
camera
mirror
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CN104293648A (en
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施宏艳
周朝明
彭金明
高云峰
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Han s Laser Technology Industry Group Co Ltd
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Han s Laser Technology Industry Group Co Ltd
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    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718Type of compounds synthesised
    • B01J2219/0072Organic compounds
    • B01J2219/00722Nucleotides

Abstract

The present invention provides a kind of gene sequencing light path system.Comprising two excitation sources in the gene sequencing light path system of present embodiment, two light sources are to work simultaneously, through the first light combination mirror, two-beam are carried out conjunction and restraint, and will be irradiated on testing sample after main spectroscopical beam combination is focused on by focusing microscope.Therefore, said gene order-checking light path system can reach to excite the visible ray of multiple specific wavelength simultaneously, improves the order-checking efficiency of said gene order-checking light path system.

Description

Gene sequencing light path system
Technical field
The present invention relates to a kind of light path system, particularly relate to a kind of gene sequencing light path system.
Background technology
In nearest 30 years, to from simple viral to the mankind and the various biologies of other mammals The decoding of the genome (coding forms of all inhereditary features) of body achieve the most progressive.For many years Various instrument is had to be developed to assist decoding or the order-checking of this genome.In view of more complicated life The extremely complicated property of the genome of object (the such as mankind), check order such genome involves time Between and cost be at a relatively high.The order-checking of first man genoid group employs 13 years and 3,500,000,000 dollars, And can incrementally expend about 300,000,000 dollars to repeat in today.But, entering in sequencing technologies and equipment Step makes to be improved accordingly on the speed in performing such order-checking and cost.In in September, 2007 public affairs The completing of second human genome of cloth consumes one-year age and 7,000 ten thousand dollars.
In gene sequencing technology field, pyrosequencing techniques (Pyrosequencing), is by Nyren et al. A kind of novel enzyme cascade sequencing technologies grown up in 1987, its repeatable and accurate performance with Sanger method gene sequencing technology compares favourably, and speed is greatly improved.Pyrosequencing techniques is by 4 Plant the enzyme cascade chemiluminescence reaction in enzymatic same reaction system.The principle of pyrosequencing techniques is: Bioluminescence is relied on to carry out the technology of gene sequencing, under the synergy of multiple enzyme, when base is correct During pairing, there is a synthetic reaction and the coupling of a chemiluminescence reaction, discharge optical signal.Optical signal Captured by high sensitivity image sensor CCD (Charged Coupled Device) in real time, be finally reached survey The purpose of sequence.Traditional light path system utilizing pyrosequencing techniques, by arranging 2 kinds of excitation sources, Alternately being scanned testing sample, alternately testing makes whole gene sequencing inefficient.
Summary of the invention
Based on this, it is necessary to provide the gene sequencing light path system that a kind of efficiency that checks order is higher.
A kind of gene sequencing light path system, luminous for the multiple base exciting testing sample, record described four Planting the luminous signal of base, described gene sequencing light path system includes:
First light source, for launching the first light beam, described first light beam can excite two kinds of base luminescences;
First expands shaping unit, is located at the side of described first light source, and described first expands shaping unit uses In the first light source is carried out shaping collimation;
Secondary light source, for launching the second light beam, described second light beam can excite other two kinds of base luminescences;
Second expands shaping unit, is located at the side of described secondary light source, and described second expands shaping unit uses In secondary light source is carried out shaping collimation;
First light combination mirror, is located at the intersection of described first light beam and described second light beam, described first light beam Reflect away from the surface of described secondary light source at described first light combination mirror, described in described second light beam transmission first Light combination mirror, described first light combination mirror is for closing restraint into beam combination by described first light beam and described second light beam;
Main spectroscope, is located at described first light combination mirror side away from described secondary light source, described main spectroscope For reflecting described beam combination;
Focusing microscope, is located at the spectroscopical side of described master, and testing sample is located at described focusing microscope Focal point, focuses on through described focusing microscope through the beam combination of described main dichroic mirror, described after focusing Beam combination is irradiated on described testing sample, and described testing sample sends the excitation beam of four kinds of wavelength, institute State focusing microscope assemble described four kinds of wavelength excitation beam become excite conjunction bundle, described in excite conjunction Shu Jing described in Main spectroscope transmission.
Wherein in an embodiment, the optical axis of described first light source is parallel with the optical axis of described secondary light source, Described gene sequencing light path system also includes that the first completely reflecting mirror, described first completely reflecting mirror are located at described second Expanding the shaping unit side away from described secondary light source, described second light beam is with described in 45 degree of incident angles First completely reflecting mirror, described second light beam incides described first through described first completely reflecting mirror reflection with 45 degree On light combination mirror.
Wherein in an embodiment, also include laser range finder and the second light combination mirror, described second light combination mirror It is located between described first light combination mirror and described main spectroscope, described laser range finder and described second light combination mirror Being oppositely arranged, the infrared laser that described laser range finder sends is vertical with described beam combination, described combination light Second light combination mirror transmission described in Shu Jing, described infrared laser closes with described beam combination through described second light combination mirror Bundle, described laser range finder is connected with described focusing microscope communication, and described laser range finder is used for measuring institute Stating the distance between focusing microscope and described object under test, described focusing microscope is according to described distance focusing.
Wherein in an embodiment, also include:
First spectroscope, is located at the described main spectroscope side away from described focusing microscope, described first point The optical axis of light microscopic optical axis spectroscopical with described master is located along the same line, and described first spectroscope is used for reflecting The described first wave length light beam closing bundle that excites, the first excitation beam of transmission residue wavelength;
First camera, is located at described first spectroscopical side, and described first wave length light beam incides described On one camera, described first camera is for recording the signal of described first wave length light beam;
Second spectroscope, is located at described first spectroscope away from the spectroscopical side of described master, described second point The optical axis of light microscopic is located along the same line with described first spectroscopical optical axis, and described second spectroscope is for anti- Penetrate the second wave length light beam of described first excitation beam, the second excitation beam of transmission residue wavelength;
Second camera, is located at described second spectroscopical side, and described second wave length light beam incides described On two cameras, described second camera is for recording the signal of described second wave length light beam;
3rd spectroscope, is located at described second spectroscope away from described first spectroscopical side, and the described 3rd Spectroscopical optical axis is located along the same line with described second spectroscopical optical axis, and described 3rd spectroscope is used for Reflect the 3rd wavelength light beam of described second excitation beam, the 3rd excitation beam of transmission residue wavelength;
Third camera, is located at the side of described second completely reflecting mirror, and described 3rd wavelength light beam incides described In third camera, described third camera is for recording the signal of described 3rd wavelength light beam;
Second completely reflecting mirror, is located at described second spectroscope away from described first spectroscopical side, and described The optical axis of two completely reflecting mirrors is located along the same line with described second spectroscopical optical axis, described second total reflection Mirror is for the 4th wavelength light beam of reflection the 3rd excitation beam;
4th camera, is located at the side of described second completely reflecting mirror, and described 4th wavelength light beam incides described On 4th camera, described 4th camera is for recording the signal of described 4th wavelength light beam.
Wherein in an embodiment, also include the first optical filtering convergence unit, described first optical filtering convergence unit Being located between described first spectroscope and described first camera, described first optical filtering convergence unit is used for filtering non- The optical signal of described first wave length light beam, described first wave length light beam through described first optical filtering convergence unit assemble, It is incident in described first camera.
Wherein in an embodiment, also include the second optical filtering convergence unit, described second optical filtering convergence unit Being located between described second spectroscope and described second camera, described second optical filtering convergence unit is used for filtering non- The optical signal of described second wave length light beam, described second wave length light beam through described second optical filtering convergence unit assemble, It is incident in described second camera.
Wherein in an embodiment, also include the 3rd optical filtering convergence unit, described 3rd optical filtering convergence unit Being located between described 3rd spectroscope and described third camera, described 3rd optical filtering convergence unit is used for filtering non- The optical signal of described 3rd wavelength light beam, described 3rd wavelength light beam through described 3rd optical filtering convergence unit assemble, It is incident in described third camera.
Wherein in an embodiment, also include the 4th optical filtering convergence unit, described 4th optical filtering convergence unit Being located between described second completely reflecting mirror and described 4th camera, described 4th optical filtering convergence unit is used for filtering The optical signal of non-described 4th wavelength light beam, described 4th wavelength light beam is through described 4th optical filtering convergence unit meeting Poly-, it is incident in described 4th camera.
Wherein in an embodiment, described first spectroscope, the second spectroscope, the 3rd spectroscope and second Completely reflecting mirror is parallel to each other placement.
Wherein in an embodiment, the distance between described first spectroscope and described first camera, described Distance between second spectroscope and described second camera, between described 3rd spectroscope and described third camera Distance, described second completely reflecting mirror and described 4th camera between distance successively decrease successively.
Comprising two excitation sources in said gene order-checking light path system, two light sources are to work simultaneously, through the Two-beam is carried out closing bundle by one light combination mirror, and will be gathered by focusing microscope through main spectroscopical beam combination It is irradiated on testing sample after Jiao.Therefore, said gene order-checking light path system can reach to excite many simultaneously Plant the visible ray of specific wavelength, improve the order-checking efficiency of said gene order-checking light path system.
Accompanying drawing explanation
Fig. 1 is the index path of the gene sequencing light path system of an embodiment.
Detailed description of the invention
For the ease of understanding the present invention, below with reference to relevant drawings, the present invention is described more fully. Accompanying drawing gives the better embodiment of the present invention.But, the present invention can come in many different forms Realize, however it is not limited to embodiments described herein.On the contrary, provide the purpose of these embodiments to be Make the disclosure is understood more thorough comprehensively.
It should be noted that when element is referred to as " being fixed on " another element, and it can be directly at another On individual element or element placed in the middle can also be there is.When an element is considered as " connection " another yuan Part, it can be directly to another element or may be simultaneously present centering elements.Used herein Term " vertical ", " level ", "left", "right" and similar statement simply to illustrate that mesh , being not offered as is unique embodiment.
Unless otherwise defined, all of technology used herein and scientific terminology and the technology belonging to the present invention The implication that the technical staff in field is generally understood that is identical.The art used the most in the description of the invention Language is intended merely to describe the purpose of specific embodiment, it is not intended that in limiting the present invention.Used herein Term " and/or " include the arbitrary and all of combination of one or more relevant Listed Items.
Referring to Fig. 1, the gene sequencing light path system 100 of present embodiment is for exciting testing sample 10 Four kinds of base luminescences, the luminous signal of four kinds of bases of record.Owing to the base type decided of testing sample 10 makes Use light source category, current technology that a kind of light source cannot be used to excite four kinds of bases to send four kinds of different wave lengths Light, so at least with two kinds of light sources.
Gene sequencing light path system 100 includes the first light source 111, secondary light source 112, first expands shaping list Unit 121, second expands shaping unit the 122, first light combination mirror 131, main spectroscope 140 and focusing microscope 150。
First light source 111 is for launching the first light beam, and the first light beam can excite a part of base luminous.Tool Body ground, the first light beam can excite two kinds of base luminescences.
First expands shaping unit 121 is located at the side of the first light source 111, and first expands shaping unit 121 For the first light source 111 is carried out shaping collimation.
Secondary light source 112 is for launching the second light beam, and the second light beam can excite other number of base luminous. Specifically, the second light beam can excite other two kinds of base luminescences.
Second expands shaping unit 122 is located at the side of secondary light source 112, and second expands shaping unit 122 For secondary light source 112 is carried out shaping collimation.
First expand shaping unit 121 and second expand shaping unit 122 emergent light can be made to become uniformly, Collimated light beam.First expand shaping unit 121 and second expand the concrete light path design of shaping unit 122 with First light source 111 and secondary light source 112 are relevant.When the semiconductor laser selecting power to compare concentration does light Source, needs to add shaping, the mirror group of even light and collimating effect;When the LED selecting relatively high power does light source, Have only to add the mirror group of even light and collimating effect.
First light combination mirror 131 is located at the first light beam and the intersection of the second light beam.First light beam closes bundle first Mirror 131 reflects away from the surface of secondary light source 112, second light beam transmission the first light combination mirror 131, and first closes bundle Mirror 131 restraints into beam combination for the first light beam and the second light beam being closed.
Main spectroscope 140 is located at first light combination mirror 131 side away from secondary light source 112, main spectroscope 140 For combinations of reflections light beam.
The side of main spectroscope 140 is located at by focusing microscope 150, and testing sample 10 is located at focusing microscope 150 Focal point, through main spectroscope 140 reflection beam combination line focus microscope 150 focus on, group after focusing Closing light beam to be irradiated on testing sample 10, testing sample 10 sends the excitation beam of four kinds of wavelength, focuses on aobvious Micro mirror 150 is assembled the excitation beam of four kinds of wavelength and is become to excite conjunction bundle, excites conjunction Shu Jingzhu spectroscope 140 transmission.
Owing to the material of the base of testing sample 10 is different, it is therefore desirable to the first light source 111 and secondary light source The wavelength of 112 is different, and the wavelength of the excitation beam ejected also is different.Specifically, the first light source The wavelength of 111 light beams sent is 532nm.The wavelength of the light beam that secondary light source 112 sends is 660nm.Four The wavelength of the excitation beam planting wavelength is respectively 558nm, 608nm, 688nm and 740nm.
The most in the present embodiment, the optical axis of the first light source 111 is parallel with the optical axis of secondary light source 112.Base Because order-checking light path system 100 also includes that the first completely reflecting mirror 161, the first completely reflecting mirror 161 are located at second and are expanded Shaping unit 122 is all-trans with 45 degree of incident angles first away from the side of secondary light source 112, the second light beam Penetrating mirror 161, the second light beam reflects through the first completely reflecting mirror 161 and incides with 45 degree on the first light combination mirror 131, First light beam and the second light beam close bundle through the first light combination mirror 131.
It is appreciated that the first completely reflecting mirror 161 can omit, the optical axis of the first light source 111 and secondary light source The optical axis of 112 is vertical, and the second light beam needs not move through the first completely reflecting mirror 161 and reflects, and can hang down with the first light beam Straight crossing.
Comprising two excitation sources in said gene order-checking light path system 100, two light sources are to work simultaneously, Carry out closing bundle by two-beam through the first light combination mirror 131, and will pass through through the beam combination of main spectroscope 140 Focusing microscope is irradiated on testing sample after focusing on 150.Therefore, said gene order-checking light path system 100 Can reach to excite the visible ray of multiple specific wavelength simultaneously, improve said gene order-checking light path system 100 Order-checking efficiency.
The gene sequencing light path system 100 of present embodiment also includes laser range finder 170 and the second light combination mirror 132.Laser range finder 170 is connected with focusing microscope 150 communication, and laser range finder 170 is used for measuring poly- Distance between focusing microscope 150 and object under test, focusing microscope 150 is according to distance focusing.Second closes Bundle mirror 132 is located between the first light combination mirror 131 and main spectroscope 140, and laser range finder 170 and second closes Bundle mirror 132 is oppositely arranged.The infrared laser that laser range finder 170 sends is vertical with beam combination, combines light Bundle is through the second light combination mirror 132 transmission.Infrared laser closes bundle through the second light combination mirror 132 reflection with beam combination.
Owing to testing sample 10 is smaller, focusing microscope 150 can realize precisely according to laser range finder 170 Focusing, improves the signal to noise ratio gathering signal, improves the accuracy of detection data.
The gene sequencing light path system 100 of present embodiment also includes first spectroscope the 141, second spectroscope 142, the 3rd spectroscope the 143, second completely reflecting mirror 162, first camera 181, second camera the 182, the 3rd Camera 183 and the 4th camera 184.
First spectroscope 141 is located at the main spectroscope 140 side away from focusing microscope 150, the first light splitting The optical axis of mirror 141 is located along the same line with the optical axis of main spectroscope 140, and the first spectroscope 141 is for anti- Penetrate and excite the first wave length light beam closing bundle, the first excitation beam of transmission residue wavelength.First wave length light beam institute An imaging optical path is become in light path.
First camera 181 is located at the side of the first spectroscope 141, and first wave length light beam incides first camera On 181, first camera 181 is for recording the signal of first wave length light beam.
Second spectroscope 142 is located at first spectroscope 141 side away from main spectroscope 140, the second light splitting The optical axis of mirror 142 and the optical axis of the first spectroscope 141 are located along the same line, and the second spectroscope 142 is used for Reflect the second wave length light beam of the first excitation beam, the second excitation beam of transmission residue wavelength.Second wave length Light beam place light path becomes an imaging optical path.
Second camera 182 is located at the side of the second spectroscope 142, and second wave length light beam incides second camera On 182, second camera 182 is for recording the signal of second wave length light beam.
Second spectroscope 142 side away from the first spectroscope 141 is located at by 3rd spectroscope 143, the 3rd point The optical axis of light microscopic 143 and the optical axis of the second spectroscope 142 are located along the same line, and the 3rd spectroscope 143 is used In reflecting the 3rd wavelength light beam of the second excitation beam, the 3rd excitation beam of transmission residue wavelength.3rd ripple Long light beam place light path becomes an imaging optical path.
Third camera 183 is located at the side of the second completely reflecting mirror 162, and the 3rd wavelength light beam incides third phase On machine 183, third camera 183 is for the signal of record the 3rd wavelength light beam.
It is appreciated that main spectroscope the 140, first spectroscope the 141, second spectroscope 142 and the 3rd spectroscope 143 is the spectroscope being coated with certain filter membrane, and the excitation beam of respective wavelength can be made to reflect, other wavelength Excitation beam transmission, so that the laser beam of respective wavelength filters out, removes spuious light beam.
Second completely reflecting mirror 162 is located at second spectroscope 142 side away from the first spectroscope 141, and second The optical axis of completely reflecting mirror 162 and the optical axis of the second spectroscope 142 are located along the same line, the second completely reflecting mirror 162 for reflecting the 4th wavelength light beam of the 3rd excitation beam.4th wavelength light beam place light path becomes an one-tenth As light path.
The side of the second completely reflecting mirror 162 is located at by 4th camera 184, and the 4th wavelength light beam incides the 4th phase On machine 184, the 4th camera 184 is for the signal of record the 4th wavelength light beam.
The most in the present embodiment, first spectroscope the 141, second spectroscope the 142, the 3rd spectroscope 143 And second completely reflecting mirror 162 be parallel to each other placement.Further, the first spectroscope 141 and first camera 181 it Between distance, distance, the 3rd spectroscope 143 and between the second spectroscope 142 and second camera 182 Distance between three cameras 183, the distance between the second completely reflecting mirror 162 and the 4th camera 184 are passed successively Subtract, make said gene order-checking light path system 100 spatially be evenly distributed so that it is compact conformation, volume is relatively Little.
The most in the present embodiment, said gene order-checking light path system 100 also includes that list is assembled in the first optical filtering Unit's the 191, second optical filtering convergence unit the 192, the 3rd optical filtering convergence unit 193 and the 4th optical filtering convergence unit 194. First optical filtering convergence unit 191 is located between the first spectroscope 141 and first camera 181.Second optical filtering meeting Poly-unit 192 is located between the second spectroscope 142 and second camera 182.3rd optical filtering convergence unit 193 It is located between the 3rd spectroscope 143 and third camera 183.It is complete that 4th optical filtering convergence unit 194 is located at second Between speculum 162 and the 4th camera 184.
First optical filtering convergence unit 191 is for filtering the optical signal of non-first wave length light beam, first wave length light beam Assemble through the first optical filtering convergence unit 191, be incident in first camera 181.
Second optical filtering convergence unit 192 is for filtering the optical signal of non-second wave length light beam, second wave length light beam Assemble through the second optical filtering convergence unit 192, be incident in second camera 182.
3rd optical filtering convergence unit 193 is for filtering the optical signal of non-3rd wavelength light beam, the 3rd wavelength light beam Assemble through the 3rd optical filtering convergence unit 193, be incident in third camera 183.
4th optical filtering convergence unit 194 is for filtering the optical signal of non-4th wavelength light beam, the 4th wavelength light beam Assemble through the 4th optical filtering convergence unit 194, be incident in the 4th camera 184.
It is appreciated that first optical filtering convergence unit the 191, second optical filtering convergence unit the 192, the 3rd filters to assemble Unit 193 and the 4th optical filtering convergence unit 194 can be assembled by convex lens group, by filter group Or a certain wavelength light is filtered by spectroscope group.First optical filtering convergence unit the 191, second optical filtering convergence unit 192, the 3rd optical filtering convergence unit 193 and the 4th optical filtering convergence unit 194 can also be for optical filtering imaging and focusing mirrors.
It is appreciated that first optical filtering convergence unit the 191, second optical filtering convergence unit the 192, the 3rd filters to assemble Unit 193 and the 4th optical filtering convergence unit 194 can omit one or more.
Send two light beams according to the first light source 111 and secondary light source 112, expand shaping list by first respectively Unit 121 and second expands shaping unit 122, makes the standard that the first light beam and the second light beam become uniformly, amplify Collimated optical beam, then pass sequentially through the first total reflective mirror, the first light combination mirror 131 and the second light combination mirror 132, arrive main point Light microscopic 140, via focusing microscope 150, is focused to high-energy-density by the first light beam and the second light beam simultaneously Hot spot, shine directly on testing sample 10, illumination path completes.Testing sample 10 is by intense light irradiation Penetrate, electron transition occurs, sends the light of four kinds of wavelength, the excitation beam of these four wavelength and the first light source 111 Mixing with secondary light source 112 veiling glare such as grade, non-signal light is filtered by line focus microscope 150, main spectroscope 140 Fall, retain four kinds of wavelength signals light.Again through first spectroscope the 141, second spectroscope the 142, the 3rd spectroscope 143 and second total reflective mirror by completely separable for the light of four kinds of wavelength, the light beam Ge Zou mono-road imaging of each wavelength Road, then assemble respectively through first optical filtering convergence unit the 191, second optical filtering convergence unit the 192, the 3rd optical filtering Unit 193 and the 4th optical filtering convergence unit 194, be imaged on the most respectively highly sensitive first camera 181, On second camera 182, third camera 183 and the 4th camera 184, capture a kind of ripple of each imaging optical path Long optical signal.
In said gene order-checking light path system 100, comprise the first light source 111 and secondary light source 112, and First light source 111 and secondary light source 112 can work simultaneously, can excite the visible ray of four kinds of wavelength, signal The imaging optical path gathered is four, the light beam imaging of every a kind of wavelength of confession.Article four, imaging optical path gathers simultaneously The light of four kinds of wavelength, each optical filtering convergence unit only carries out specifying a kind of wavelength, reduce optical element design and Difficulty of processing, reduces equipment cost.When gathering every kind of wavelength channels, first camera 181, second camera 182, all can obtain signal clearly on third camera 183 and the 4th camera 184, improve and finally gather signal Signal to noise ratio.
Embodiment described above only have expressed the several embodiments of the present invention, and it describes more concrete and detailed, But therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that, for this area Those of ordinary skill for, without departing from the inventive concept of the premise, it is also possible to make some deformation and Improving, these broadly fall into protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be with appended Claim is as the criterion.

Claims (10)

1. a gene sequencing light path system, luminous for the multiple base exciting testing sample, record is described The luminous signal of four kinds of bases, it is characterised in that described gene sequencing light path system includes:
First light source, for launching the first light beam, described first light beam can excite a part of base luminous;
First expands shaping unit, is located at the side of described first light source, and described first expands shaping unit uses In the first light source is carried out shaping collimation;
Secondary light source, for launching the second light beam, described second light beam can excite other number of base luminous;
Second expands shaping unit, is located at the side of described secondary light source, and described second expands shaping unit uses In secondary light source is carried out shaping collimation;
First light combination mirror, is located at the intersection of described first light beam and described second light beam, described first light beam Reflecting away from the surface of described secondary light source at described first light combination mirror, described second light beam closes through described first Bundle mirror transmission, described first light combination mirror is for closing restraint into combination light by described first light beam and described second light beam Bundle;
Main spectroscope, is located at described first light combination mirror side away from described secondary light source, described main spectroscope For reflecting described beam combination;
Focusing microscope, is located at the spectroscopical side of described master, and testing sample is located at described focusing microscope Focal point, focuses on through described focusing microscope through the beam combination of described main dichroic mirror, described after focusing Beam combination is irradiated on described testing sample, and described testing sample sends the excitation beam of four kinds of wavelength, institute State focusing microscope assemble described four kinds of wavelength excitation beam become excite conjunction bundle, described in excite conjunction Shu Jing described in Main spectroscope transmission.
Gene sequencing light path system the most according to claim 1, it is characterised in that described first light source Optical axis parallel with the optical axis of described secondary light source, described gene sequencing light path system also includes the first total reflection Mirror, described first completely reflecting mirror is located at described second and expands the shaping unit side away from described secondary light source, Described second light beam is with the first completely reflecting mirror described in 45 degree of incident angles, and described second light beam is through described first Completely reflecting mirror reflection is incided on described first light combination mirror with 45 degree.
Gene sequencing light path system the most according to claim 1, it is characterised in that also include Laser Measuring Distance meter and the second light combination mirror, described second light combination mirror is located between described first light combination mirror and described main spectroscope, Described laser range finder is oppositely arranged with described second light combination mirror, the infrared laser that described laser range finder sends Vertical with described beam combination, described beam combination is through described second light combination mirror transmission, described infrared laser warp Described second light combination mirror and described beam combination close bundle, described laser range finder and described focusing microscope communication Connecting, described laser range finder is used for measuring the distance between described focusing microscope and described object under test, Described focusing microscope is according to described distance focusing.
4. according to the gene sequencing light path system described in claims 1 to 3 any one, it is characterised in that also Including:
First spectroscope, is located at the described main spectroscope side away from described focusing microscope, described first point The optical axis of light microscopic optical axis spectroscopical with described master is located along the same line, and described first spectroscope is used for reflecting The described first wave length light beam closing bundle that excites, the first excitation beam of transmission residue wavelength;
First camera, is located at described first spectroscopical side, and described first wave length light beam incides described On one camera, described first camera is for recording the signal of described first wave length light beam;
Second spectroscope, is located at described first spectroscope away from the spectroscopical side of described master, described second point The optical axis of light microscopic is located along the same line with described first spectroscopical optical axis, and described second spectroscope is for anti- Penetrate the second wave length light beam of described first excitation beam, the second excitation beam of transmission residue wavelength;
Second camera, is located at described second spectroscopical side, and described second wave length light beam incides described On two cameras, described second camera is for recording the signal of described second wave length light beam;
3rd spectroscope, is located at described second spectroscope away from described first spectroscopical side, and the described 3rd Spectroscopical optical axis is located along the same line with described second spectroscopical optical axis, and described 3rd spectroscope is used for Reflect the 3rd wavelength light beam of described second excitation beam, the 3rd excitation beam of transmission residue wavelength;
Third camera, is located at the side of described second completely reflecting mirror, and described 3rd wavelength light beam incides described In third camera, described third camera is for recording the signal of described 3rd wavelength light beam;
Second completely reflecting mirror, is located at described second spectroscope away from described first spectroscopical side, and described The optical axis of two completely reflecting mirrors is located along the same line with described second spectroscopical optical axis, described second total reflection Mirror is for the 4th wavelength light beam of reflection the 3rd excitation beam;
4th camera, is located at the side of described second completely reflecting mirror, and described 4th wavelength light beam incides described On 4th camera, described 4th camera is for recording the signal of described 4th wavelength light beam.
Gene sequencing light path system the most according to claim 4, it is characterised in that also include the first filter Light convergence unit, described first optical filtering convergence unit is located between described first spectroscope and described first camera, Described first optical filtering convergence unit is for filtering the optical signal of non-described first wave length light beam, described first wave length Light beam is assembled through described first optical filtering convergence unit, is incident in described first camera.
Gene sequencing light path system the most according to claim 5, it is characterised in that also include the second filter Light convergence unit, described second optical filtering convergence unit is located between described second spectroscope and described second camera, Described second optical filtering convergence unit is for filtering the optical signal of non-described second wave length light beam, described second wave length Light beam is assembled through described second optical filtering convergence unit, is incident in described second camera.
Gene sequencing light path system the most according to claim 6, it is characterised in that also include the 3rd filter Light convergence unit, described 3rd optical filtering convergence unit is located between described 3rd spectroscope and described third camera, Described 3rd optical filtering convergence unit is for filtering the optical signal of non-described 3rd wavelength light beam, described 3rd wavelength Light beam is assembled through described 3rd optical filtering convergence unit, is incident in described third camera.
Gene sequencing light path system the most according to claim 7, it is characterised in that also include the 4th filter Light convergence unit, described 4th optical filtering convergence unit be located at described second completely reflecting mirror and described 4th camera it Between, described 4th optical filtering convergence unit for filtering the optical signal of non-described 4th wavelength light beam, the described 4th Wavelength light beam is assembled through described 4th optical filtering convergence unit, is incident in described 4th camera.
Gene sequencing light path system the most according to claim 4, it is characterised in that described first light splitting Mirror, the second spectroscope, the 3rd spectroscope and the second completely reflecting mirror are parallel to each other placement.
Gene sequencing light path system the most according to claim 9, it is characterised in that described first point Distance between light microscopic and described first camera, distance between described second spectroscope and described second camera, Distance between described 3rd spectroscope and described third camera, described second completely reflecting mirror and described 4th phase Distance between machine is successively decreased successively.
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