CN1237187C - DNA probe array synthesis, photoswitch array combination and manufacture thereof - Google Patents
DNA probe array synthesis, photoswitch array combination and manufacture thereof Download PDFInfo
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- CN1237187C CN1237187C CN 02103696 CN02103696A CN1237187C CN 1237187 C CN1237187 C CN 1237187C CN 02103696 CN02103696 CN 02103696 CN 02103696 A CN02103696 A CN 02103696A CN 1237187 C CN1237187 C CN 1237187C
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Abstract
The present invention relates to an optical switch array assembly for optical guide synthesis of a DNA probe array and optical excitation detection of a hybridization DNA probe array, which comprises a wafer, an optical switch array, a driving circuit, an addressing circuit, an optical beam guide hole and a DNA array, wherein optical switches work in a Fabry-Perot cavity mode; the cavity is formed by micro-machinery processing and composed of two parellel mirrors separated by an air layer and medium thin films. An optical beam is irradiated in the synthesis position on a glass sheet after the optical switches are selectively started, and thereby, DNA probes are synthesized by optical guide. After the optical switches are selectively started, the optical beam is irradiated on the hybridization DNA probe arranged on the glass sheet to excite the hybridization DNA probe to emit fluorescence, and thereby, the optical detection is carried out by the hybridization DNA probe.
Description
1. invention field
This patent relates generally to dna probe array light guiding array of photoswitch synthetic and optical excitation detection usefulness and makes up, be particularly related to by micromachined and form, can selective actuation, and be used for the array of photoswitch combination that the guiding of dna probe array light is synthetic and optical excitation is surveyed.
2. prior art is described
Follow engineered developing rapidly, the gene diagnosis of disease is day by day popularized, and discloses the Basic Ways that biological phenomena becomes life science especially by genetic analysis.An urgent demand of gene diagnosis and genetic analysis to develop as early as possible exactly simple in structure, tolerance range height, time saving and energy saving, cheap DNA sequencing instrument.The DNA chip that occurs recent years has that volume is little, and sample and reagent consumption are few, and measuring speed is fast, can be by the advantages such as personnel operation without special training.
The dna probe array has adopted the method for light guiding to synthesize.For example, adopt the nucleoside phosphoramidate (nucleosidephosphoramidite) " building block " of 5 ' terminal protection by light guiding synthetic oligonucleotide.5 ' not the group of end protection can be the light mutability, also can be acid-labile.In this building-up process, synthesising part is former to be added on protection, can protection remove by illumination.Only after synthesising part was removed protection, different types of mononucleotide molecule just can be coupled to synthesising part and get on.The synthetic of different sequence dna probes realized by repeatedly circulating of this removal protection and coupling.Three kinds of light guiding synthetic methods commonly used are arranged, and these three kinds of methods are respectively: adopt the variable blocking group of light and use up direct removal blocking group; Adopt 4 of acid-labile, 4 '-dimethoxytrityl (dimethoxytrityl or DMT) blocking group is removed the photoresist material that is used as protective layer with using up; Adopt the DMT blocking group and use up the light mutability polymeric film of removing as protective layer (PAG).
This several method all needs to use many being similar to and makes the used technology of unicircuit.Wherein use to such an extent that the maximum technology of number of times is figure transfer or photoetching technique.This processing requirement designs the photoetching masterplate earlier, makes it comprise a large amount of regularly arranged some districts.When source of parallel light is shone substrate by the photoetching masterplate, can produce similar figure at substrate surface.It is synthetic to have only light-struck some district that DNA just takes place, and the some district that does not have rayed does not then synthesize.
Adopt the synthetic not homotactic dna probe array of this method need carry out repeatedly photoetching.For example, the probe array of making 25 nucleic acid glycosides needs to carry out 100 photoetching at least, because the sequence of each probe all is not quite similar, the photoetching masterplate that each photoetching is used is also all inequality.
The photoetching technique of writing direct is used to improve the existing cost of aforesaid method synthesized dna probe array, problems such as quality and efficient.This writing direct carried out dna probe array synthetic method without the fixed Lithographic template, but generates figure temporarily, therefore needs the motion graphics maker.System with this direct imaging function includes spatial light modulator, and this modulator is controlled by computer, can both provide a predefined figure when making each photoetching, and projecting substrate surface, to carry out the light guiding synthetic.
Known spatial light modulator sub microcomputer tool modulator and microelectromechanicpositioning modulator.It is the microreflection lens array that a kind of micromechanics modulator is arranged, and with the micro metal speculum reflected light is modulated, and makes a branch of light optionally reflex to specific zone.A kind of micromachine modulator is arranged, and what its operation was relied on is not the modulation reflected light, but the modulation transmitted light, for example liquid-crystal display just belongs to this micromachine modulator.
There are some problems in this method of writing direct the photoetching synthesized dna probe.Mainly be that this system must configuration realize light path aligned mechanism, with the specific region on the beam alignment substrate, these mechanisms comprise X-Y translation stage and step motor, and the distance that platform is moved equals the centre compartment of specific region on the substrate.Will spend a lot of labours and time with this clock method mass production dna probe array, so production cost is very high.In addition, along with the scale of probe array constantly increases, production cost climbs up and up especially.
Secondly, the optical detection of hybrid dna probe also requires a cover complicated mechanical devices.This device comprises sample table, light source, photo-detector, scanning device and optical repeater etc.Scanning device is wanted mobile example platform and optical repeater simultaneously.Optical repeater will be followed the tracks of between zero position of surveying and final position.So that each or a group probe can both accurately be located, and can measure the fluorescence that it sends under beam excitation.
3. summary of the invention
Aim of the present invention generally speaking, will solve above-mentioned dna probe array light exactly and guide the problem that exists in synthetic and the hybrid dna probe array optical excitation Detection Techniques.Particularly, to reach following several purpose exactly.
First purpose is developed a kind of array of photoswitch combination exactly, can be used to carry out the synthetic and hybrid dna probe array optical excitation detection of light guiding of dna probe array, and light path is aimed at and the mechanism of probe tracking and do not need to dispose.
Second purpose developed the combination of a kind of array of photoswitch exactly, and its array of photoswitch and driving circuit thereof are integrated on the same substrate, and needs are carried out each group position of dna probe synthetic can both the selective actuation photoswitch, introduce light beam and shone.
The 3rd purpose, develop a kind of array of photoswitch combination exactly, its array of photoswitch and driving circuit thereof are integrated on the same substrate, and needs are carried out the dna probe of each hybridization of optical excitation fluorescence detection can both the selective actuation photoswitch, introduce light beam and shone.
The 4th purpose, develop a kind of array of photoswitch combination exactly, its array of photoswitch and driving circuit thereof are integrated on the same substrate, make all need the position of synthesized dna probe can both start photoswitch, introduce light beam and directly shine, and can therefrom not pass and produce any interference owing to light path to reaction soln.
The 5th purpose, develop a kind of array of photoswitch combination exactly, its array of photoswitch and driving circuit thereof are integrated on the same substrate, make the dna probe of all hybridization can both start photoswitch, introduce light beam and directly shine, and can therefrom not pass and produce any interference owing to light path to adjacent hybridization probe.
The 6th purpose developed a kind of array of photoswitch combination of being made up of array of photoswitch and driving circuit exactly, can produce in batches with sophisticated integrated circuit technique of industrial production and micromachining technology.
The 7th purpose developed a kind of dna probe light that is used for exactly and guided array of photoswitch combination synthetic and that the optical excitation of hybrid dna probe is surveyed, and integrated with pilot circuit, the realization light path is aimed at automatically, and production cost is low, and travelling speed is fast, and operation is simple.
Last purpose, develop a kind of array of photoswitch combination that the guiding of dna probe light is synthetic and the optical excitation of hybrid dna probe is surveyed that is used for exactly, do not need molecular biological experiment condition, do not need senior specialized personnel to participate in person, can be placed in any laboratory, hospital, and clinic are by general technician's operation.
To achieve these goals and other purpose, the array of photoswitch combined feature structure that the present invention proposes comprises the silicon chip as substrate, be in the array of photoswitch within the silicon chip, be installed on the sheet glass on the silicon chip, and the dna probe array that is in glass sheet surface.Be integrated with the driving circuit that starts photoswitch in the silicon chip, also may be integrated with the addressing circuit of choosing photoswitch.Silicon chip back is carved with many deep holes, and each deep hole is aimed at a photoswitch, when photoswitch is opened, can guide a branch of light to project the glass sheet surface of photoswitch top.
Every photoswitch of array of photoswitch combination is all parallel by the two sides, and is made up of the isolated film mirror of gas cloud, also promptly is made up of Fabry-Perot (Fabry-Perot) cavity.According to known theory, the transmission coefficient t of loss-free Fabry-Perot cavity is the reflection R of mirror
1And R
2And the function of airspace h between minute surface:
T=[(1-R
1) (1-R
2)]/{ [1-(R
1R
2)
1/2]
2+ 4 (R
1R
2)
1/2Sin
2(2 π h/ λ) } wherein λ be the work optical wavelength.
Sinusoidal value in the following formula denominator is respectively 0 and at 1 o'clock, and the numerical value of this formula reaches maximum value and minimum value respectively.Therefore, when the h value was λ/4 several times, transmissivity became
T
Max=[(1-R
1) (1-R
2)]/[1-(R
1R
2)
1/2]
2For h=0, λ/2, λ
T
Min=[(1-R
1) (1-R
2)]/[1+ (R
1R
2)
1/2]
2For h=λ/4,3 λ/4
This shows that when the reflection coefficient value of two sides mirror equated, the transmissivity of Fabry-Perot cavity was always 1, and reflection coefficient is always 0.Therefore, obtain maximum contrast, or maximum Fabry-Perot cavity transmissivity and minimum Fabry-Perot cavity reflection coefficient, the reflection R of two sides film mirror
1And R
2To equate as far as possible and get bigger numerical value as far as possible.
Basic theories is told us simultaneously, when the thickness value of mirror is λ/(4n)) odd-multiple the time, the reflection R of mirror
1And R
2Get maximum value, wherein n is the specific refractory power of mirror material.In the case, the R of mirror
1And R
2Be expressed as
R
1，2＝[(n
2-n
0)/(n
2+n
0)]
2
N wherein
0Specific refractory power for the mirror primer.
The film mirror of array of photoswitch combination of the present invention preferably is made of unformed carbonization silicon.Its first replacement scheme is to be made of the silicon nitride that plasma reinforced chemical vapour deposition (PECVD) method forms.Its second replacement scheme is to be made of the silicon nitride that low-pressure chemical vapor deposition (LPCVD) method forms.For UV-irradiation, be that silicon carbide or silicon nitride all are transparent, can not produce the absorption loss of light.
According to bibliographical information, the specific refractory power of unformed carbonization silicon is 2.48 to 2.65, and the specific refractory power of silicon nitride is 2.0 to 2.1.For the film of freely supporting, medium is an air under it, and n is arranged
0=1.Formula with above-mentioned reflection coefficient calculates, and draws the reflection R of unformed carbonization silicon mirror
1,2Be respectively 52% and 56%, and the reflection R of silicon nitride mirror
1,2Be respectively 40% and 36%.
Photoswitch based on the Fabry-Perot cavity is operated by electrostatic force.The top mirror of Fabry-Perot cavity is by four elastic beam supports.When no impressed voltage imposes on cavity, and h is when equaling the odd-multiple of λ/4, and the transmissivity of cavity is got maximum value, and this moment, switch was in closing condition.Do the time spent as impressed voltage, under electrostatic forcing, elastic beam bends, and the top mirror of cavity pulls to the bottom, changes the numerical value of h.In case the numerical value of h equals the even-multiple of λ/4, the transmissivity of cavity is then got minimum value, and this moment, switch entered opening state.When light source vertical irradiation silicon chip back, each deep hole in the silicon chip all can be guided Ray Of Light into the photoswitch corresponding with this deep hole.When photoswitch was in closing condition, the light beam that deep hole is introduced was gone back by vertical reflection.When photoswitch was in opening state, the light beam that deep hole is introduced passed switch, moves on along original Pass Fabric direction.
Impressed voltage imposes on selected photoswitch group by driving circuit and addressing circuit decision.Driving circuit comprises analog switch and gating circuit, and addressing circuit comprises shift-register circuit etc.These two kinds of circuit can be partly or entirely and array of photoswitch be integrated on the same silicon chip.
Light guiding with array of photoswitch combination carrying out dna probe array of the present invention is synthetic, and as adopting DMT blocking group and photoresist process, its processing procedure is as follows.The first, the design dna probe sequence, and write the computer program of per step illumination.Computer program is used to control the distribution of array of photoswitch Push And Release, and to form specific light and shade alternative dot pattern, each corresponding needs in some district carries out dna probe synthetic position.Second; sheet glass Biao Mian Tu in the array of photoswitch combination covers photoresist material; and with combination pack into reaction bench or reaction chamber; so that spray the mononucleotide solution that contains the DMT blocking group, mononucleotide has VITAMIN B4 (A), cytosine(Cyt) (C); guanine (G); four kinds of thymus pyrimidines (T), each only with a kind of, and different number of times is with different mononucleotides.Photoresist material can be positive glue, also can be negative glue, and its effect is to make the zone of removing protection accept illumination or avoid illumination respectively.The 3rd, start photoswitch, open Jupiter, the irradiation synthetic surface through after the regular hour, is closed Jupiter again.The 4th, carry out photoresist developing and corrosion, and with the exposed sheet glass zone of acid treatment, to remove the blocking group on the synthetic oligonucleotide, remove remaining photoresist material then.The 5th, allow the mononucleotide that contains the DMT blocking group be coupled on the oligonucleotide of removing protection.The 7th, the sheet glass Shang Tu in the array of photoswitch combination covers photoresist material once more.Above steps is carried out successively repeatedly, till the dna probe array that synthesizes multiple required DNA series.
The analytical procedure of dna probe array comprises, with all compositions of fluorescent-substance markers DNA sample; Sample is ejected on the sheet glass, carries out specific hybrid with dna probe array on the sheet glass.If certain composition of sample is successful with some probe hybridization on the sheet glass, this composition will be fixed on corresponding probe area.If be opened with the corresponding photoswitch in this zone, then the dna probe of this area hybridization sends fluorescence under ultraviolet excitation.Send the address information in zone according to fluorescence,, and then can determine the DNA sequence of sample with regard to the corresponding probe type of deducibility.
4. accompanying drawing summary
Figure 1A represents that a photoswitch of array of photoswitch combination of the present invention is unlocked, and introduces the upper surface that Ray Of Light shines sheet glass thus, thereby forms a dna probe at this place.
Figure 1B represents that a photoswitch of array of photoswitch combination of the present invention is opened, and introduces the dna probe that Ray Of Light shines a hybridization of glass sheet surface thus, and the dna probe of hybridizing under the excitation of this light is launched fluorescence.
Fig. 2 represents to make up from array of photoswitch of the present invention the part excision skeleton view of a repeating unit that cuts down, this unit comprises a photoswitch, be in a deep hole under the photoswitch, be in a sheet glass on the photoswitch and a dna probe that forms in glass sheet surface.
Fig. 3 is for being combined in the cross sectional representation after manufacturing step 1 is finished from array of photoswitch of the present invention, the upper surface that the figure shows at a silicon chip has formed first silicon dioxide layer, the first unformed carbonization silicon layer, second silicon dioxide layer and the second unformed carbonization silicon layer.
Fig. 4 the figure shows and form electric connection line above silicon chip, isolating bar, and reflectance coating for be combined in the cross sectional representation after manufacturing step 2 is finished from array of photoswitch of the present invention.
Fig. 5 has formed the groove of filling for being combined in the cross sectional representation after manufacturing step 3 is finished from array of photoswitch of the present invention, the figure shows above silicon chip.
Fig. 6 the figure shows the plane configuration that depicts the Fabry-Perot cavity above silicon chip for be combined in the cross sectional representation after manufacturing step 4 is finished from array of photoswitch of the present invention.
Fig. 7 has formed the deep hole of aiming at Fabry-Perot cavity plane configuration for being combined in the cross sectional representation after manufacturing step 5 is finished from array of photoswitch of the present invention, the figure shows below silicon chip.
Fig. 8 the figure shows the photoswitch based on Fabry-Perot cavity operating mode that forms behind second dioxide layer in eroding Fabry-Perot cavity plane configuration district for be combined in the cross sectional representation after manufacturing step 6 is finished from array of photoswitch of the present invention.
Fig. 9 the figure shows the top that a sheet glass places the Fabry-Perot cavity for be combined in the cross sectional representation after manufacturing step 7 is finished from array of photoswitch of the present invention.
Figure 10 the figure shows the upper surface that a dna probe is formed at sheet glass for be combined in the cross sectional representation after manufacturing step 8 is finished from array of photoswitch of the present invention.
5. optimum implementation explanation
Array of photoswitch combination of the present invention is shown in Figure 1A and Figure 1B.The array of photoswitch that Figure 1A represents constitutes the silicon chip 201 that comprises as substrate, be arranged in silicon chip 201 some photoswitches that are in closing condition by 102 signs, be arranged in silicon chip 201 some photoswitches that are in opened condition by 103 signs, place the sheet glass 104 of silicon chip 201 tops, by 112 signs, be in the dna probe of sheet glass 104 upper surfaces with some.Array of photoswitch constitutes and also comprises some electronic switch by 107 signs, by 108 vertical transfer registers that indicate with by 109 horizontal shifting registers that indicate.Each photoswitch is controlled by an electronic switch, and the electronic switch of opening is united selected by vertical transfer register and horizontal shifting register.Some deep holes are carved with at the back side of silicon chip 201, photoswitch of each deep hole aiming.One ultraviolet source vertical irradiation silicon chip, 201 back sides are arranged, and be divided into some light beams that separate by deep hole.When photoswitch was in closing condition, as 102 marks, the light beam in the deep hole was gone back by the photoswitch vertical reflection, as 106 marks.When photoswitch was in opened condition, as 103 marks, the light beam in the deep hole can pass photoswitch, and the direction of broadcasting along original Pass moves on, and projected the surface of sheet glass 104, as 105 marks.Be covered with the DNA resulting solution by the glass sheet surface of light beam irradiates such as Tu, it is synthetic that the light guiding can take place, and forms dna probe, as 112 signs.
The combination of the represented array of photoswitch of Figure 1B, except replace labels 112 with label 113, other label is all identical with label connotation among Figure 1B.Label 113 expression dna probes 112 and the hybrid dna probe that combines back formation with fluorescein-labeled DNA sample specificity.Under uv light induction, the dna probe of hybridization is understood emitting fluorescence, and is detected by photo-detector.If the dna probe type of hybridization is known, also promptly the dna probe of hybridization order is known, can infer according to the address information of the photoswitch of opening this sample DNA type or this sample DNA order.
The periodicity of array of photoswitch combination of the present invention repeats the unit as shown in Figure 2.This unit comprises silicon chip 201, is deposited on the first unformed carbonization silicon layer 202 on the silicon chip 201 successively, the silicon dioxide layer 203 and the second unformed carbonization silicon layer layer 204, and a Fabry-Perot cavity.This Fabry-Perot cavity is by the first unformed carbonization silicon fiml 205, and the air buffer 206 and the second unformed carbonization silicon fiml 207 are formed.The second unformed carbonization silicon fiml 207 is supported by the unformed carbonization silicon beam 208 of four symmetrical distributions.Every unformed carbonization silicon beam 208 links to each other with silicon chip by two spud piles 209.Metal electrode 210 covers peripheral part of whole unformed carbonization silicon beam 208 and unformed carbonization silicon fiml 207.The back side of the first unformed carbonization silicon fiml 205 is covered by the film 211 that prevents luminous reflectance, and aims at the deep hole 213 that opens wide to silicon chip 201 back sides.The zone that the Fabry-Perot cavity occupies is encircled by eight protection against corrosion walls 214.Every the spud pile 209 protection against corrosion wall 214 corresponding with links to each other.The silicon chip back side is coated with optical reflection film 215.One sheet glass 217 places silicon chip 201 tops by parting bead 216.A dna probe 218 is in sheet glass 217 upper surfaces, and aims at deep hole 213 in the bottom.
The first and second unformed carbonization silicon fimls 205 and 207 thickness setting are λ/(4n
SiC) odd-multiple so that the reflection coefficient of these two unformed carbonization silicon fimls is all got maximum value, wherein n
SiCSpecific refractory power for unformed carbonization silicon.The original thickness of air buffer 206 is set at the odd-multiple of λ/4, so that the transmissivity minimalization of Fabry-Perot cavity, or make this cavity be in the total reflection state.
The resistivity of unformed carbonization silicon is up to 10
10To 10
14Therefore ohm-cm by the slab construction of electrode 210 with the silicon chip combination, can become good parallel plate capacitor.When a voltage put on this electrical condenser, the electrostatic forcing of generation was in unformed carbonization silicon beam 208, made it crooked and unformed carbonization silicon fiml 207 is passed downwards, thereby reduced the thickness of air buffer 206.And when the thickness of air buffer 206 was reduced to the even-multiple that equals λ/4, the Fabry-Perot cavity was driven into the total transmissivity state.
Dna probe 218 forms by the light beam irradiates guiding is synthetic, and its shape and size are replicated in deep hole 213 substantially, because illumination beam is introduced by deep hole, also promptly by deep hole, splits from the parallel rays at irradiation silicon chip 201 back sides.
As a kind of replacement scheme, the Fabry-Perot cavity is made of the PECVD silicon nitride film.In the case, replaced by the multilayered structure that PECVD silicon nitride layer-aluminium lamination-the PECVD silicon nitride layer is formed by the multilayered structure that unformed carbonization silicon layer-silicon dioxide layer-unformed carbonization silicon layer is formed.
As another kind of replacement scheme, the Fabry-Perot cavity is made of the lpcvd silicon nitride film.In the case, replaced by the multilayered structure that the lpcvd silicon nitride layer-titanium dioxide osmanthus layer-the lpcvd silicon nitride layer is formed by the multilayered structure that unformed carbonization silicon layer-silicon dioxide layer-unformed carbonization silicon layer is formed.
The preferential photoswitch assemblage characteristic manufacturing process of recommending of the present invention comprises eight steps altogether, and each step is represented by Fig. 3 to Figure 10 respectively.First manufacturing step is ready to the silicon chip 301 as substrate as shown in Figure 3, and deposits first silicon dioxide layer, 302, the first unformed carbonization silicon layer 303, the second silicon dioxide layers 304 and the second unformed carbonization silicon layers 305 successively at this silicon chip surface.Silicon chip 301 can live through the MOS circuit manufacture procedure, and is manufactured with some dissimilar MOS circuit, comprises analog electronic switching circuit, gating circuit, addressing circuit etc.First silicon dioxide layer is deposited by PECVD, and its thickness is less times greater than λ/(4n
SiO2), make that this layer is subjected to slight corrosion in forming the deep hole process after a while after, its thickness just in time equals λ/(4n
SiO2).The deposition parameter that is adopted is 250 watts of RF power, 300 degrees centigrade of temperature, N
2O flow 300 standard cubic centimeters per minute, SiH
4Flow 40 standard cubic centimeters per minute and air pressure 240 millitorrs.With this understanding the sedimentation rate of Chan Shenging be 600 dusts/minute.Also by the PECVD deposition, its thickness equals λ/(4n to the first unformed carbonization silicon layer 303
SiC) odd-multiple.The cvd furnace that is adopted is a double frequency RF process furnace.The deposition parameter that is adopted is air pressure 600 millitorrs, SiH
4Flow 10 standard cubic centimeters per minute, CH
4Flow 250 standard cubic centimeters per minute, Ar flow 300 standard cubic centimeters per minute, 60 watts of 300 degrees centigrade of temperature and RF power.The stress of the unformed carbonization silicon layer 303 that is obtained under these conditions is lower than 50 milli handkerchiefs, and has stronger corrosion resistance.Also by above-mentioned silicon-dioxide pecvd process deposition, sedimentary thickness is the odd-multiple of λ/4 to second silicon dioxide layer 304.Also by above-mentioned unformed carbonization silicon pecvd process deposition, its thickness also equals λ/(4n to the second unformed carbonization silicon layer 305
SiC) odd-multiple, but sedimentary thickness is greater than the thickness of unformed carbonization silicon layer 303, so that this layer has than higher physical strength.
Second manufacturing step forms electric connection line 306 respectively on the two sides of silicon chip 301 as shown in Figure 4, isolating bar 308 and reflectance coating 307.For this reason, the aluminium layer deposition that the deposited by electron beam evaporation technology is thick with 1500 dusts is to the two sides of silicon chip 301.Aluminium lamination to upper surface carries out photoetching corrosion, to form the aluminium lamination figure, aluminium corrosion H
3PO
4: HAC: CH
3COOH=8: 0.5: 0.1: 1 etchant solution.Upper surface at silicon chip 301 forms the litho pattern that another only exposes parting bead 308 then, the aluminium lamination of another layer of deposition 3 micron thickness on the photoresist material figure.Remove the photoresist material under the aluminium lamination,, only preserve the aluminium lamination on the parting bead 308 in company with the aluminium lamination of removing on the photoresist material.With this aluminium lamination thickening technology the aluminium lamination of parting bead 308 is thickeied greater than 3 microns, other aluminum layer thickness everywhere of silicon chip surface then remains unchanged.
The 3rd manufacturing step forms the corrosion stop slot 309 and the spud pile (not marking among the figure) of filling as shown in Figure 5.This step is included in the second unformed carbonization silicon layer 305 and second silicon dioxide layer 304 and etches dead slot.For this reason, on the second unformed carbonization silicon layer 305, form litho pattern earlier, then silicon chip is put into reactive ion etching (RIE) device and carried out dry etching.Etching condition is 60 watts of power, air pressure 0.05 millitorr, CF
4Flow 70 standard cubic centimeters per minute, SF
6Flow 10 standard cubic centimeters per minute, and O
2, flow 10 standard cubic centimeters per minute.With this understanding, the erosion rate of generation be 800 dusts/minute, generation select coefficient to be respectively 1.2 and 0.7 to silicon-dioxide with to the corrosion of photoresist material.Second silicon dioxide layer 304 is by the HF solution corrosion of dilution.The dead slot that carves the most at last tamps again, for this reason, protects the All Ranges except that dead slot earlier with photoresist, deposits unformed silicon layer with the PECVD method then, to cover the dead slot inwall, tamps dead slot with PECVD method deposition of silica layer at last.The molten photoresist material that goes then except that the dead slot of filling, is all removed along with the photoresist material dissolving at the unformed silicon and the silicon dioxide layer of other area deposition.
The 4th manufacturing step depicts the two dimensional structure of Fabry-Perot cavity as shown in Figure 6.After forming photoresist material protection figure, the second unformed carbonization silicon layer 305 is carried out dry etching, the corrosive condition is identical with the technology of the unformed carbonization silicon of above-mentioned corrosion.After this step finished, the groove 309 of filling became corrosion stop wall 309.
The 5th manufacturing step carves deep hole 311 at the back side of silicon chip 301 shown in 7.Before the etching, form 9 to 10 micron thickness, have the litho pattern of corrosion window at the back side of silicon chip 301.Then remove aluminium lamination in the window, put Bo Si (Bosch) dry etching device again into, with the silicon layer in the corrosion window with the aluminium etchant solution.Corrosion is performed until silicon chip 301 corrosion saturating, stops at first silicon dioxide layer 302 of another side at last.The diameter of deep hole 311 is 4 to 30 microns, and representative value is 10 microns.The silicon dioxide layer 302 that expose deep hole 311 bottoms is because of the slight corrosion of process, and thickness reduces to some extent, but can be used as anti-reflective film 312 just.
The 6th manufacturing step falls to corrode second silicon dioxide layer, 304 parts that stop wall 309 is surrounded with the HF solution corrosion that dilutes shown in 8.Form thin 313, four the amorphous carborundum beams 314 of first amorphous carborundum thus, the air buffer 315 and second amorphous carborundum thin 316.Two relative edges of every amorphous carborundum beam 314 are by two spud pile supports, and this spud pile is to form in the manufacturing processed of filling slot 309.First amorphous carborundum thin 313 is of a size of 4 * 4 to 40 * 40 square microns, and representative value is 20 * 20 square microns.The length of amorphous carborundum beam 314 and widely be respectively 4 to 30 microns, representative value is 10 microns, 1 to 5 micron, representative value is 2 microns.
The 7th manufacturing step places sheet glass 317 on the silicon chip 301 shown in 9, separates by parting bead 308 therebetween.The back side of sheet glass 317 can cover anti-reflective film, and its back side periphery also can seal, so that protect array of photoswitch in the building-up process of DNA.Sheet glass 317 is disposable, and array of photoswitch then can be reused, and therefore, the mode of sealing should be removed.
The 8th manufacturing step is shown in 10, at sheet glass 317 surperficial synthesized dna probe arrays 318.The variable blocking group of the synthetic employing light of dna probe array 318 is removed protective layer technology with using up.This processing requirement is at first used up variable blocking group sheet glass 317 surfaces is modified.Introduce beam bundles by opening first photoswitch then, make it shine first and need carry out DNA synthetic position, to produce the reaction hydroxyl.Then first kind 3 of Pen Tu ' end be activated 5 ' terminal hydroxy group be not combined with the deoxynucleotide of the variable group of light, makes in synthesising position generation coupling.After coupling and oxide treatment, again sheet glass 317 is washed, open second batch of photoswitch then and introduce beam bundles, make second batch of palpus of its irradiation carry out the synthetic position.Then ' end is activated 5 ' terminal hydroxy group is not combined with the deoxynucleotide Pen Tu of the variable group of light to synthesising position with second kind 3.This selective light removes protection and the coupling circulation repeats down, till the dna probe array that obtains desired sequence.
As a kind of replacement scheme, the guiding of the light of dna probe array 318 is synthetic adopts acid easy 4,4 ' not hold blocking group and photoresist process.
As another kind of replacement scheme, the synthetic DMT of employing of the light of dna probe array 318 guiding blocking group and the polymer thin membrane process that contains the sour survivor of producing of light (PAG).
Second preferential manufacturing photoswitch combined feature flow process of the present invention adopts by first aluminium lamination, the one PECVD silicon nitride layer, second aluminium lamination, multilayered structure with the 2nd PECVD silicon nitride layer composition, to replace by first silicon dioxide layer, the first unformed carbonization silicon layer, the multilayered structure that second silicon dioxide layer and the second unformed carbonization silicon layer are formed.The difference of two kinds of processing procedures specifically comprises following several:
1. the used parameter of deposition PECVD silicon nitride is: SiH
4Flow 170 standard cubic centimeters per minute, NH
3Flow 30 standard cubic centimeters per minute, 300 degrees centigrade of temperature, air pressure 450 millitorrs, 250 watts of power, generation sedimentation rate 160 dusts/minute.The parameter of RIE corrosion PECVD silicon nitride layer: CHF
4Flow 7.5 standard cubic centimeters per minute, N
2Flow 42.5 standard cubic centimeters per minute, air pressure 37.5 millitorrs, 60 watts of power, generation erosion rate 250 dusts/minute.
2. fill the material silicon-dioxide of dead slot.
3. carving first aluminium lamination that exposes behind the deep hole need erode, and deposits a bed thickness λ/(4n then
SiO2) silicon dioxide layer as anti-reflective film.
The 3rd preferential manufacturing photoswitch combined feature flow process of the present invention adopts by first silicon dioxide layer, the first lpcvd silicon nitride layer, second silicon dioxide layer, multilayered structure with second lpcvd silicon nitride layer composition, to replace by first silicon dioxide layer, the first unformed carbonization silicon layer, the multilayered structure that second silicon dioxide layer and the second unformed carbonization silicon layer are formed.The difference of two kinds of processing procedures specifically comprises following several:
1. when array of photoswitch began to make, the MOS circuit on the silicon chip was not finished metallization step as yet.
2.RIE the used parameter of corrosion lpcvd silicon nitride is: CHF
4Flow 7.5 standard cubic centimeters per minute, N
2Flow 42.5 standard cubic centimeters per minute, air pressure 37.5 millitorrs, 60 watts of power, generation erosion rate 250 dusts/minute.
3. metallization forms electrical wiring, and isolating bar and reflectance coating are used as the final step of array of photoswitch processing procedure to carry out.
The above array of photoswitch based on the Fabry-Perot cavity is an illustration of the present invention, and content of the present invention also not exclusively is confined to this.Above-mentioned illustration is carried out some revise, know well these professional engineering technical personnel and after reading this explanation, be easy to accomplish.Therefore marrow of the present invention is not to be limited by foregoing, but is limited by claim of the present invention.
Claims (10)
1. array of photoswitch combination that is used for dna probe light compositing and hybrid dna probe optical excitation detection, its feature structure comprises:
One silicon chip,
One is in the silicon chip, parallel by the two sides, and is made up of the film mirror of air insulated, and with the array of photoswitch of Fabry-Perot cavity mode work,
One is in the silicon chip, by electronic switch with comprise that the gating circuit of vertical transfer register and horizontal shifting register forms, and the driving circuit that impels photoswitch to move,
One is in the deep hole array in the silicon chip, and each deep hole leads to a photoswitch, and tells a branch of light from the UV-light of irradiation silicon chip, makes it be transmitted to the photoswitch place along deep hole,
One place the sheet glass that there is anti-reflection film at the back side on the silicon chip and
One is on the sheet glass, by light guiding synthetic DNA array.
2. array of photoswitch combination that is used for dna probe light compositing and hybrid dna probe optical excitation detection, its feature structure comprises:
One silicon chip,
One is in the silicon chip, parallel by the two sides, and is made up of the film mirror of air insulated, and with the array of photoswitch of Fabry-Bai Luo cavity mode work,
One is in the silicon chip, by electronic switch with comprise that the gating circuit of vertical transfer register and horizontal shifting register forms, and the driving circuit that impels photoswitch to move,
One is in the deep hole array in the silicon chip, and each deep hole leads to a photoswitch, and tells a branch of light from the UV-light of irradiation silicon chip, makes it be transmitted to the photoswitch place along deep hole,
One place the sheet glass that there is anti-reflection film at the back side on the silicon chip and
One is in the hybrid dna array on the sheet glass.
3. method that is used to make the array of photoswitch combination that claim 1 and 2 described dna probe light compositings and the optical excitation of hybrid dna probe survey, its feature manufacturing step comprises:
Be ready to be manufactured with analog electronic switching circuit, the silicon chip of gating circuit and addressing circuit,
The four-level membrane structure of formation of deposits Fabry-Perot cavity on silicon chip, wherein the first layer is used for deep hole corrosion stop layer, and the second layer and the 4th layer are used for forming speculum, form air buffer behind the 3rd layer of erosion removal,
The technology of deposited by electron beam evaporation aluminium film forms electric connection line on the four-level membrane structure, isolating bar, and light reflective film,
In four-layer structure, be formed by etching groove, and deposit unformed silicon and silica membrane groove is filled forming corrosion stop slot and spud pile,
Four-level membrane to the superiors corrodes, and with the superstructure of forming method Fabry-Perot-type cavity, and makes corrosion stop slot on every side become the corrosion stop wall,
From silicon chip back side corrosion deep hole, make the first layer film of its bottom, straight-through silicon chip front with rich this dry etching device, and make the center of its centrally aligned front etch figure square,
The first layer film is transformed into anti-reflective film,
The second layer film of Fabry-Perot cavity superstructure bottom is removed in selective corrosion, makes it be transformed into air buffer, thus form the two-face mirror that constitutes by the second layer and the 4th boundary layer film and therebetween the Fabry one Perot cavity formed of air buffer and
Above silicon chip, place the sheet glass that backside deposition has anti-reflective film.
4. be used for the method that light that dna probe light compositing and the optical excitation of hybrid dna probe survey is opened array combination according to the described manufacturing of claim 3, its feature manufacturing step also comprise described second and the four-level membrane material be unformed carbonization silicon layer.
5. be used for the method that light that dna probe light compositing and the optical excitation of hybrid dna probe survey is opened array combination according to the described manufacturing of claim 3, its feature manufacturing step also comprise described second and the four-level membrane material be the PECVD silicon nitride layer.
6. be used for the method that light that dna probe light compositing and the optical excitation of hybrid dna probe survey is opened array combination according to the described manufacturing of claim 3, its feature manufacturing step also comprise described second and the four-level membrane material be the lpcvd silicon nitride layer.
7. be used for the method that light that dna probe light compositing and the optical excitation of hybrid dna probe survey is opened array combination according to the described manufacturing of claim 3, its feature manufacturing step comprises that also described three-layer thin-film material is the PECVD silicon dioxide layer.
8. be used for the method that light that dna probe light compositing and the optical excitation of hybrid dna probe survey is opened array combination according to the described manufacturing of claim 3, its feature manufacturing step comprises that also described three-layer thin-film material is the aluminium lamination of electron beam evaporation.
9. be used for the method that light that dna probe light compositing and the optical excitation of hybrid dna probe survey is opened array combination according to the described manufacturing of claim 3, its feature manufacturing step also comprises makes it be transformed into anti-reflection film the first layer silica membrane attenuate.
10. be used for the method that light that dna probe light compositing and the optical excitation of hybrid dna probe survey is opened array combination according to the described manufacturing of claim 3, its feature manufacturing step also comprises deposit the layer of silicon dioxide film again as anti-reflection film after will the first layer aluminium thin film corrosive removing.
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