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Ultra-compact (54 × 58 × 8.5 mm) uye yakafara-aperture (1 × 7 mm) mipfumbamwe-mavara spectrometer yakagadzirwa, "yakakamurwa nepakati" nemhando gumi yezvionioni zvedichroic, izvo zvakashandiswa pakarepo spectral imaging.Chiitiko chechiedza chinoyerera chine chikamu chemuchinjiko chidiki pane aperture saizi yakakamurwa kuita tambo inoenderera 20 nm yakafara uye mapfumbamwe mavara fluxes ane epakati wavelengths e530, 550, 570, 590, 610, 630, 650, 670 uye 690 nm.Mapikicha emapfumbamwe emavara hova anoyerwa panguva imwe chete zvinoyerwa neiyo mufananidzo sensor.Kusiyana neyakajairwa dichroic girazi arrays, iyo yakagadziridzwa dichroic girazi array ine yakasarudzika maviri-zvidimbu zvigadziriso, izvo kwete chete kuwedzera huwandu hwemavara anogona kuyerwa panguva imwe chete, asi zvakare inovandudza chifananidzo chega chega rukova.Iyo yakagadziridzwa mipfumbamwe-mavara spectrometer inoshandiswa mana-capillary electrophoresis.Kuongororwa kwehuwandu hwemadhayi masere panguva imwe chete mucapillary imwe neimwe achishandisa mapfumbamwe-mavara laser-induced fluorescence.Sezvo iyo mipfumbamwe-mavara spectrometer isiri chete-idiki-diki uye isingadhure, asi zvakare ine yakakwira inopenya flux uye yakakwana spectral resolution yeakawanda spectral imaging application, inogona kushandiswa zvakanyanya munzvimbo dzakasiyana siyana.
Hyperspectral uye multispectral imaging yave chikamu chakakosha chekuongorora nyeredzi2, kure kure nekuonekwa kwePasi3,4, kutonga kwemhando yekudya nemvura5,6, kuchengetedza art uye archaeology7, forensics8, surgery9, biomedical analysis uye diagnostics10,11 etc. Munda 1 Unyanzvi hunokosha hunodiwa ,12,13.Nzira dzekuyera huwandu hwechiedza chinoburitswa nenzvimbo yega yega yekubuda mumunda wekuona dzakakamurwa kuita (1) point scanning ("broom")14,15, (2) linear scanning ("panicle")16,17,18 , (3) kureba kunoongorora masaisai19,20,21 uye (4) mifananidzo22,23,24,25.Panyaya yenzira dzese idzi, kugadzirisa kwenzvimbo, spectral resolution uye kugadzirisa kwechinguva kune hukama hwekutengesa9,10,12,26.Pamusoro pezvo, kubuda kwechiedza kune zvakakosha pakunzwa, kureva chiratidzo-kune-ruzha reshiyo mukufungidzira kwe spectral26.Kupenya kwechiedza, ndiko kuti, kugona kwekushandisa chiedza, kwakanyatso kuenzana nereshiyo yechiyero chaicho chakayerwa chechiedza chenzvimbo imwe neimwe inopenya pachikamu chenguva kusvika kuhuwandu hwechiedza cheyero yewavelength yakayerwa.Category (4) inzira yakakodzera apo kusimba kana kuwanda kwechiedza chinoburitswa nenzvimbo yega yega yekuburitsa kuchichinja nekufamba kwenguva kana kana nzvimbo yenzvimbo imwe neimwe yekuburitsa ichichinja nekufamba kwenguva nekuti mwenje wechiedza chinoburitswa nezvose zvinobuda zvinoyerwa panguva imwe chete.24.
Dzakawanda dzenzira dziri pamusoro dzakasanganiswa nehombe, yakaoma uye/kana inodhura spectrometers vachishandisa 18 gratings kana 14, 16, 22, 23 prisms yemakirasi (1), (2) uye (4) kana 20, 21 mafirita disks, mafirita emvura. .Crystalline tunable mafirita (LCTF)25 kana acousto-optic tunable mafirita (AOTF)19 echikamu (3).Kusiyana neizvi, chikamu (4) multi-mirror spectrometers idiki uye isingadhuri nekuda kwekugadzirisa kwayo kuri nyore27,28,29,30.Pamusoro pezvo, ivo vane kupenya kwakakwira nekuti mwenje wakagovaniswa negirazi yega yega dichroic (kureva kuti, mwenje unopfuudzwa uye unotaridzwa wechiedza chechiitiko pagirazi rega rega re dichroic) rinoshandiswa zvizere uye rinoramba richishandiswa.Zvisineyi, huwandu hwemabhendi ewavelength (kureva mavara) hunofanirwa kuyerwa panguva imwe chete hunogumira kuanenge mana.
Spectral imaging yakavakirwa pakuongorora kwefluorescence inowanzoshandiswa pakuongorora kwakawanda mukuongorora kwebiomedical uye diagnostics 10, 13.Mukuwandisa, sezvo maanalytes akawanda (semuenzaniso, DNA chaiyo kana mapuroteni) akanyorwa nedhayi dzakasiyana dzefluorescent, muongorori wega wega aripo pane imwe neimwe nzvimbo yekubuda mundima yekuona inoverengerwa uchishandisa multicomponent ongororo.32 inopwanya iyo yakaonekwa fluorescence spectrum inoburitswa neimwe emission point.Munguva iyi, dhayi dzakasiyana, imwe neimwe ichiburitsa fluorescence yakasiyana, inogona kugadzikana, ndiko kuti, kugarisana munzvimbo nenguva.Parizvino, huwandu hwepamusoro hwedhayi hunogona kunakidzwa neiyo imwe laser beam ndeye masere33.Uyu muganho wepamusoro hautemerwe ne spectral resolution (kureva, nhamba yemavara), asi nehupamhi hwe fluorescence spectrum (≥50 nm) uye huwandu hwedhayi Stokes shift (≤200 nm) paFRET (uchishandisa FRET)10 .Zvisinei, nhamba yemavara inofanira kuva yakakura kudarika kana kuenzana nenhamba yedhayi kuti ibvise spectral kupindirana kwemavara akasanganiswa31,32.Naizvozvo, zvinodikanwa kuwedzera nhamba yemavara akayerwa panguva imwe chete kusvika masere kana kupfuura.
Munguva pfupi yapfuura, iyo Ultra-compact heptachroic spectrometer (ichishandisa akatevedzana heptychroic magirazi uye mufananidzo sensor kuyera mana fluorescent fluxes) yakagadzirwa.Iyo spectrometer ine maviri kusvika matatu maodha ehukuru idiki pane yakajairwa spectrometers uchishandisa gratings kana prisms34,35.Zvisinei, zvakaoma kuisa magirazi edichroic anopfuura manomwe mu spectrometer uye panguva imwe chete kuyera anopfuura mavara manomwe36,37.Nekuwedzera kwenhamba yezvioni zvedichroic, mutsauko mukuru mukureba kwenzira dzemaziso e dichroic light fluxes inowedzera, uye zvinova zvakaoma kuratidza chiedza chese pane imwe ndege inonzwika.Kureba kwenzira yechiedza chechiedza chinoyerera zvakare inowedzera, saka hupamhi hwe spectrometer aperture (kureva hupamhi hwepamusoro hwechiedza chakaongororwa ne spectrometer) hunoderera.
Mukupindura kumatambudziko ari pamusoro apa, ultra-compact nine-color spectrometer ine maviri-layer "dichroic" decachromatic mirror array uye sensor yemufananidzo wepakarepo spectral imaging [chikamu (4)] yakagadzirwa.Kuenzaniswa neyakapfuura ma spectrometers, iyo yakagadziridzwa spectrometer ine mutsauko mudiki muhupamhi hwepamusoro nzira yehurefu uye hudiki hurefu hwehurefu hwenzira.Yakaiswa kune ina-capillary electrophoresis kuona laser-induced mapfumbamwe-ruvara fluorescence uye kuverengera kutama panguva imwe chete kwemadhayi masere mucapillary yega yega.Sezvo iyo yakagadziridzwa spectrometer isiri chete yakanyanya-diki uye isingadhure, asi zvakare ine yakakwira inopenya flux uye yakakwana spectral resolution yeakawanda spectral imaging application, inogona kushandiswa zvakanyanya munzvimbo dzakasiyana siyana.
Yechinyakare mipfumbamwe-mavara spectrometer inoratidzwa mufig.1a.Magadzirirwo ayo anotevera eiyo yapfuura ultra-diki-diki-mavara manomwe-mavara spectrometer 31. Iyo ine mapfumbamwe dichroic magirazi akarongwa akatwasuka pakona ye45 ° kurudyi, uye mufananidzo sensor (S) iri pamusoro mapfumbamwe dichroic magirazi.Chiedza chinopinda kubva pazasi (C0) chinopatsanurwa nemhando yezvioni zvipfumbamwe zvedichroic kuita zvipfumbamwe zvinoyerera zvichikwira (C1, C2, C3, C4, C5, C6, C7, C8 uye C9).Ese pfumbamwe nzizi dzemavara anodyiswa zvakananga kune sensor yemufananidzo uye anoonekwa panguva imwe chete.Muchidzidzo ichi, C1, C2, C3, C4, C5, C6, C7, C8, uye C9 zviri muhurongwa hwewavelength uye zvinomiririrwa nemagenta, violet, bhuruu, cyan, girinhi, yero, orenji, tsvuku-orenji, uye. dzvuku, zvichiteerana.Kunyange zvazvo mavara aya anoshandiswa mugwaro rino, sezvinoratidzwa mumufananidzo 3, nokuti anosiyana nemavara chaiwo anoonekwa neziso remunhu.
Schematic dhayagiramu yezvakajairwa uye mitsva mipfumbamwe-mavara spectrometers.(a) Chitarisiko chemavara mapfumbamwe chine ruzhinji rwezvionioni zvipfumbamwe zvedichroic.(b) New-color spectrometer ine maviri-layer dichroic mirror array.Chiitiko mwenje flux C0 yakakamurwa kuita mapfumbamwe ane mavara emwenje fluxes C1-C9 uye akaonekwa nemufananidzo sensor S.
Iyo yakagadziridzwa mitsva mipfumbamwe-mavara spectrometer ine maviri-layer dichroic mirror grating uye mufananidzo sensor, sezvakaratidzwa muFig. 1b.Muchikamu chepasi, magirazi mashanu edichroic akarereka 45 ° kurudyi, anoenderana kurudyi kubva pakati peiyo dhizaini yedecamers.Padanho repamusoro, magirazi mashanu ekuwedzera edichroic akarereka 45 ° kuruboshwe uye anowanikwa kubva pakati kuenda kuruboshwe.Chionioni chekuruboshwe chechikamu chepasi uye chekurudyi dichroic girazi chepamusoro chinopindirana.Chiitiko mwenje flux (C0) yakakamurwa kubva pazasi kuita ina inobuda chromatic fluxes (C1-C4) neshanu dichroic magirazi kurudyi uye mashanu anobuda chromatic fluxes (C5-C4) neshanu dichroic magirazi kuruboshwe C9).Kufanana neakajairwa mapfumbamwe-mavara spectrometers, ese mapfumbamwe mavara hova anopinzwa zvakananga mumufananidzo sensor (S) uye anoonekwa panguva imwe chete.Kuenzanisa Figures 1a uye 1b, munhu anogona kuona kuti munyaya itsva mapfumbamwe-ruvara spectrometer, zvose musiyano mukuru uye kureba optical nzira kureba mipfumbamwe ruvara fluxes vari hafu.
Kuvakwa kwakadzama kweiyo ultra-diki-diki-maviri-dichroic mirror array 29 mm (upamhi) × 31 mm (yakadzika) × 6 mm (kureba) inoratidzwa muMufananidzo 2. The decimal dichroic mirror array ine zvioni zvishanu zvedichroic kurudyi. (M1-M5) uye mashanu dichroic magirazi kuruboshwe (M6-M9 uye imwe M5), imwe neimwe dichroic girazi inogadziriswa mupamusoro aluminium bracket.Ese magirazi edichroic anodzika kuti atsivise kufambisa kwakafanana nekuda kwekudzokororwa kwekuyerera kuburikidza nemagirazi.Pazasi peM1, bhendi-pass sefa (BP) inogadziriswa.M1 uye BP zviyero 10mm (rutivi rurefu) x 1.9mm (rutivi rupfupi) x 0.5mm (ukobvu).Zviyero zvezvioni zvedichroic zvakasara 15 mm × 1.9 mm × 0.5 mm.Iyo matrix pitch pakati peM1 neM2 i1.7 mm, nepo matrix pitch yemamwe magirazi edichroic ari 1.6 mm.Pamusoro pemuonde.2c inosanganisa chiitiko mwenje flux C0 uye mapfumbamwe ane mavara emwenje anoyerera C1-C9, akapatsanurwa ne de-chamber matrix yemagirazi.
Kuvaka maviri-layer dichroic mirror matrix.(a) Maonero ekuona uye (b) maonero e-cross-sectional maviri-layer dichroic mirror array (dimensions 29 mm x 31 mm x 6 mm).Iyo ine magirazi mashanu e-dichroic (M1-M5) ari muchikamu chepasi, magirazi mashanu e-dichroic (M6-M9 uye imwe M5) iri muchikamu chepamusoro, uye bandpass sefa (BP) iri pazasi M1.(c) Muchinjikwa-chikamu maonero munzira yakatwasuka, ine C0 uye C1-C9 inopindirana.
Upamhi hwechivharo mugwara rakachinjika, rinoratidzwa nehupamhi C0 mumufananidzo 2, c, 1 mm, uye munzira yakatarisana nendege yeFig. 2, c, yakapihwa nekugadzirwa kwealuminium bracket, - 7 mm.Ndiko kuti, itsva mipfumbamwe-mavara spectrometer ine yakakura aperture saizi 1 mm × 7 mm.Nzira yemaziso yeC4 ndiyo yakareba kupfuura yose pakati peC1-C9, uye nzira yeC4 yemaziso mukati me dichroic mirror array, nekuda kwepamusoro-soro-duku (29 mm × 31 mm × 6 mm), ndeye 12 mm.Panguva imwecheteyo, iyo optical nzira urefu hweC5 ndiyo ipfupi pakati peC1-C9, uye nzira yekuona yeC5 ndeye 5.7mm.Naizvozvo, iyo yakanyanya musiyano muOptical nzira kureba ndeye 6.3 mm.Iyo iri pamusoro peiyo nzira yekutarisa kureba inogadziriswa yeoptical nzira kureba kweiyo optical transmission yeM1-M9 uye BP (kubva kuquartz).
Iyo spectral zvivakwa zveM1−М9 uye VR zvakaverengerwa kuti fluxes С1, С2, С3, С4, С5, С6, С7, С8 uye С9 ari muwavelength renji 520-540, 540-560, 560-8580, 560-8580, 560-8580, -600 , 600-620, 620-640, 640-660, 660-680, uye 680-700 nm, maererano.
Mufananidzo weiyo yakagadzirwa matrix yemagirazi e decachromatic inoratidzwa muFig. 3a.M1-M9 neBP zvakanamirwa kune 45-degree kutsetseka uye yakatwasuka ndege yerutsigiro rwealuminium, zvichiteerana, nepo M1 neBP zvakavanzwa kumashure kwechimiro.
Kugadzirwa kwemhando dzakasiyana dzemagirazi e decan uye kuratidzwa kwayo.(a) Ruzhinji rwemagirazi akagadzirwa edecachromatic.(b) A 1 mm × 7 mm pfumbamwe-ruvara rwakapatsanurwa mufananidzo wakaiswa pabepa rakaiswa pamberi pemhando yezvionioni zve decachromatic uye nekumashure nechiedza chichena.(c) Mutsara wemagirazi edecochromatic akavhenekerwa nechiedza chichena kubva kumashure.(d) Rukova rune-ruvara rwakatsemura rupfumbamwe runobva mugirazi re decane array, inocherechedzwa nekuisa acrylic canister izere nehutsi pamberi pechionioni che decane pa c uye kusviba mukamuri.
Mucherechedzo wekutapurirana wakayerwa weM1-M9 C0 paakona yezviitiko zve45 ° uye kuyerwa kwekutapurirana kweBP C0 paakona yechiitiko che0 ° inoratidzwa muFigs.4a.Iyo yekufambisa spectra yeC1-C9 inoenderana neC0 inoratidzwa muFig.4b.Aya ma spectra akaverengerwa kubva kune spectra muFig.4a maererano nenzira ye optical C1-C9 mumufananidzo 4a.1b uye 2c.Somuenzaniso, TS(C4) = TS (BP) × [1 − TS (M1)] × TS (M2) × TS (M3) × TS (M4) × [1 − TS (M5)], TS(C9) = TS (BP) × TS (M1) × [1 − TS (M6)] × TS (M7) × TS (M8) × TS (M9) × [1 − TS (M5)], apo TS(X) uye [ 1 − TS(X)] ndiyo inotapurirana uye inoratidza spectra yeX, zvichiteerana.Sezvinoratidzwa mumufananidzo 4b, bandwidth (bandwidth ≥50%) yeC1, C2, C3, C4, C5, C6, C7, C8 uye C9 ndeye 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 uye 682-699 nm.Migumisiro iyi inopindirana nemhando dzakagadzirwa.Mukuwedzera, kushandiswa kwekushanda kweC0 chiedza kwakakwirira, kureva, avhareji yepamusoro yeC1-C9 chiedza transmittance ndeye 92%.
Transmission spectra ye dichroic mirror uye kupatsanurwa kwemavara mapfumbamwe.(a) Chiyero chekufambisa kweM1-M9 pa 45 ° chiitiko uye BP pa 0 ° chiitiko.(b) Transmission spectra yeC1-C9 inoenderana neC0 yakaverengwa kubva (a).
Pamusoro pemuonde.3c, ruzhinji rwemagirazi edichroic anowanikwa vertically, kuitira kuti rutivi rwayo rworudyi muFig. 3a irwo rutivi rwepamusoro uye bhanhire chena ye collimated LED (C0) inodzokera shure.Ruzhinji rwemagirazi e decachromatic anoratidzwa muMufananidzo 3a akaiswa mu 54 mm (kureba) × 58 mm (kudzika) × 8.5 mm (ukobvu) adapta.Pamusoro pemuonde.3d, kuwedzera kune iyo nyika inoratidzwa mufig.3c, acrylic tank yakazadzwa nehutsi yakaiswa pamberi pemagirazi edecochromatic, nemwenje wemukamuri wakadzima.Nekuda kweizvozvo, pfumbamwe hova dze dichroic dzinoonekwa mutangi, dzichibva kune akatevedzana e decatroic magirazi.Rukova rwega rwega rwakapatsanuka rune rectangular muchinjiko chikamu chine zviyero zve 1 × 7 mm, iyo inoenderana neaperture size yetsva mipfumbamwe-mavara spectrometer.Mumufananidzo 3b, bepa rinoiswa pamberi pezvionioni zvedichroic muFigure 3c, uye mufananidzo we 1 x 7 mm wemipfumbamwe dichroic hova inoratidzirwa papepa inocherechedzwa kubva munzira yekufamba kwebepa.hova.Mapfumbamwe emavara ekupatsanura hova mufig.3b uye d iC4, C3, C2, C1, C5, C6, C7, C8 neC9 kubva kumusoro kusvika pasi, iyo inogona kuonekwawo munhamba 1 uye 2. 1b uye 2c.Iwo anocherechedzwa mumavara anoenderana newavelengths adzo.Pamusana pekuderera kwechiedza chechiedza che LED (ona Supplementary Fig. S3) uye kunzwisiswa kwekamera yemuvara inoshandiswa kutora C9 (682-699 nm) muFig. Zvimwe zvinoparadzanisa zvinoyerera hazvina simba.Saizvozvowo, C9 yainyatsooneka kune ziso rakashama.Zvichakadaro, C2 (rukova rwechipiri kubva kumusoro) rinotaridzika rakasvibirira muMufananidzo 3, asi rinotarisa zvakanyanya kutsvuka kune ziso rakashama.
Kuchinja kubva kuMufananidzo 3c kusvika kune d kunoratidzwa muSupplementary Video 1. Pakarepo mushure mokunge chiedza chichena chinobva ku LED chichipfuura ne decachromatic mirror array, inoparadzana panguva imwe chete kuva mipfumbamwe mavara enzizi.Pakupedzisira, utsi huri mubhodhoro hwakapera zvishoma nezvishoma kubva kumusoro kusvika pasi, zvokuti hupfu hupfumbamwe hwemavara hwakanyangarika kubva kumusoro kusvika pasi.Kusiyana neizvi, muSupplementary Vhidhiyo 2, apo kureba kwechiitiko chechiedza chekuyerera pamhando yezvionioni zve decachromatic zvakashandurwa kubva kureba kuenda kupfupi mukurongeka kwe690, 671, 650, 632, 610, 589, 568, 550 uye 532 nm. ., Chete hova dzakapatsanurwa dzinoenderana dzenzizi pfumbamwe dzakapatsanurwa muhurongwa hweC9, C8, C7, C6, C5, C4, C3, C2, uye C1 dzinoratidzwa.Iyo acrylic reservoir inotsiviwa ne quartz dziva, uye maflakes ega ega shunted kuyerera anogona kucherechedzwa zvakajeka kubva kunotsvedza kumusoro.Uye zvakare, iyo sub-vhidhiyo 3 inogadziriswa zvekuti iyo wavelength shanduko chikamu cheiyo sub-vhidhiyo 2 inodzokororwa.Uku ndiko kutaura kwakanyanyisa kwemaitiro edecochromatic array yemagirazi.
Mhedzisiro iri pamusoro inoratidza kuti yakagadzirwa decachromatic mirror array kana iyo mipfumbamwe-ruvara spectrometer inoshanda sezvakarongwa.Iyo mipfumbamwe-mavara spectrometer inoumbwa nekukwirisa ruzhinji rwe decachromatic magirazi ane adapta akananga pamufananidzo sensor board.
Luminous flux ine wavelength range kubva ku400 kusvika 750 nm, inobudiswa neine radiation points φ50 μm, iri pa 1 mm intervals munzira inotarisana nendege yeFig. mana lenzi φ1 mm ane kureba kwe1.4 mm uye pitch ye1 mm.Hova ina collimated (ina C0) inoitika paDP yeiyo mipfumbamwe-mavara spectrometer, yakaparadzana pa 1 mm nguva.Rudzi rwemagirazi edichroic anokamura rukova rumwe norumwe (C0) kuita nzizi pfumbamwe dzemavara (C1-C9).Mahova makumi matatu nenhanhatu anozobuda (maseti mana eC1-C9) anobva aiswa jekiseni akananga muCMOS (S) mufananidzo sensor yakabatana zvakananga kune akatevedzana edichroic magirazi.Somugumisiro, sezvakaratidzwa muFig. 5a, nekuda kwediki madiki optical path difference uye pfupi pfupi yepamusoro optical nzira, mifananidzo yevose 36 hova yakaonekwa panguva imwe chete uye zvakajeka nehukuru hwakafanana.Zvinoenderana neiyo yakadzika spectra (ona Supplementary Figure S4), iwo mufananidzo kusimba kwemapoka mana C1, C2 uye C3 yakaderera.Mifananidzo makumi matatu nenhanhatu yaive 0.57 ± 0.05 mm muhukuru (zvinoreva ± SD).Saka, kukwidziridzwa kwemufananidzo kwakawedzera 11.4.Kupatsanurana kwakatwasuka pakati pemifananidzo kunosvika 1 mm (spacing yakafanana neyerenzi yelenzi) uye yakachinjika kupatsanuka inoita maavhareji 1.6 mm (yakafanana mirairi yegirazi redhichroic).Nekuti saizi yemufananidzo idiki pane chinhambwe chiri pakati pemifananidzo, mufananidzo wega wega unogona kuyerwa wakazvimirira (une low crosstalk).Zvichakadaro, mifananidzo yemakumi maviri nesere nzizi yakanyorwa neyakajairwa manomwe-mavara spectrometer yakashandiswa muchidzidzo chedu chekare inoratidzwa muFig. 5 B. Mutsara wezvioni zvinomwe zve dichroic zvakagadzirwa nekubvisa maviri ekurudyi dichroic magirazi kubva kune array mapfumbamwe dichroic. magirazi muMufananidzo 1a.Haisi mifananidzo yese yakapinza, saizi yemufananidzo inowedzera kubva kuC1 kuenda kuC7.Mifananidzo makumi maviri nesere iri 0.70 ± 0.19 mm muhukuru.Nokudaro, zvakaoma kuchengetedza mufananidzo wepamusoro mumifananidzo yose.The coefficient of variation (CV) yemufananidzo saizi 28 muFigure 5b yaive 28%, ukuwo CV yemufananidzo saizi 36 paFigure 5a yakadzikira kusvika 9%.Migumisiro iri pamusoro inoratidza kuti itsva mipfumbamwe-mavara spectrometer haingowedzeri nhamba yemavara akayerwa panguva imwe chete kubva pamanomwe kusvika kumapfumbamwe, asi inewo mufananidzo wepamusoro wemuvara wega wega.
Kuenzanisa kwemhando yemufananidzo wakapatsanurwa wakaumbwa neakajairika uye matsva ma spectrometers.(a) Mapoka mana emifananidzo mipfumbamwe yakapatsanurwa (C1-C9) inogadzirwa neiyo mipfumbamwe-mavara spectrometer.(b) Maseti mana emifananidzo yakapatsanurwa ine mavara manomwe (C1-C7) akaumbwa neyakajairwa-mavara manomwe-mavara spectrometer.Fluxes (C0) ine wavelengths kubva ku400 kusvika 750 nm kubva kune ina emission point inosanganiswa uye chiitiko pane yega spectrometer, zvichiteerana.
Mamiriro emhando ye9-color spectrometer akaongororwa nekuedza uye mhedzisiro yekuongorora inoratidzwa muMufananidzo 6. Cherechedza kuti Mufananidzo 6a unoratidza mhedzisiro yakafanana neFigure 5a, kureva pawavelengths ye4 C0 400-750 nm, mifananidzo yese ye36 inoonekwa. (4 mapoka C1-C9).Kusiyana neizvi, sezvinoratidzwa mumufananidzo 6b-j, kana C0 imwe neimwe iine wavelength chaiyo ye530, 550, 570, 590, 610, 630, 650, 670, kana 690 nm, pane inenge ina chete mifananidzo inoenderana (ina. mapoka akaonekwa C1, C2, C3, C4, C5, C6, C7, C8 kana C9).Zvisinei, mamwe emifananidzo ari pedyo nemifananidzo mina inoenderana inonyatsoonekwa nokuti C1-C9 transmission spectra inoratidzwa muFig. 4b inopindirana zvishoma uye C0 imwe neimwe ine 10 nm band pane imwe wavelength sezvakatsanangurwa nenzira.Iyi mhedzisiro inoenderana neC1-C9 transmission spectra inoratidzwa muFig.4b uye mavhidhiyo ekuwedzera 2 uye 3. Mune mamwe mazwi, mapfumbamwe mavara spectrometer anoshanda sezvinotarisirwa zvichienderana nemigumisiro inoratidzwa mufig.4b.Naizvozvo, zvinogumiswa kuti mufananidzo wakanyanya kugovera C1-C9 ndiyo inotaridzika yeC0 yega yega.
Spectral maitiro ezvipfumbamwe-mavara spectrometer.Iyo mipfumbamwe-mavara-mavara spectrometer inogadzira mana seti yemipfumbamwe-mavara akapatsanurwa mifananidzo (C1-C9) kana chiitiko mwenje (ina C0) ine wavelength ye (a) 400-750 nm (sezvinoratidzwa muFigure 5a), (b) 530 nm.nm, (c) 550 nm, (d) 570 nm, (e) 590 nm, (f) 610 nm, (g) 630 nm, (h) 650 nm, (i) 670 nm, (j) 690 nm, zvichiteerana.
Iyo yakagadziridzwa mipfumbamwe-mavara spectrometer yakashandiswa mana-capillary electrophoresis (kuti uwane ruzivo, ona Supplementary Materials)31,34,35.Iwo mana-capillary matrix ane mana capillaries (kunze dhayamita 360 μm uye mukati dhayamita 50 μm) iri pa 1 mm intervals panzvimbo ye laser irradiation.Samples ine zvidimbu zveDNA zvakanyorwa ne8 dhayi, zvinoti FL-6C (dhayi 1), JOE-6C (dhayi 2), dR6G (dhayi 3), TMR-6C (dhayi 4), CXR-6C (dhayi 5), TOM- 6C (dhayi 6), LIZ (dhayi 7), uye WEN (dhayi 8) mukukwira kurongeka kwefluorescent wavelength, yakapatsanurwa mune yega yega capillaries ina (inozonzi Cap1, Cap2, Cap3, uye Cap4).Laser-induced fluorescence kubva kuCap1-Cap4 yakasanganiswa nerudungwe rwemalenzi mana uye panguva imwe chete yakarekodhwa ine mipfumbamwe-mavara spectrometer.Kusimba kwemasimba emapfumbamwe-mavara (C1-C9) fluorescence panguva ye electrophoresis, kureva, mapfumbamwe-mavara electrophoregram ye capillary imwe neimwe, inoratidzwa muFig. 7a.Iyo yakaenzana nepfumbamwe-mavara electrophoregram inowanikwa muCap1-Cap4.Sezvinoratidzwa neCap1 miseve muMufananidzo 7a, nhongonya sere pane imwe neimwe-yemavara-mavara electrophoregram inoratidza imwe fluorescence emission kubva Dye1-Dye8, zvichiteerana.
Panguva imwe chete kuverengwa kwedhayi sere uchishandisa mipfumbamwe-mavara mana-capillary electrophoresis spectrometer.(a) Mapfumbamwe-ruvara (C1-C9) electrophoregram yega yega capillary.Iwo manhongonya masere anoratidzwa nemiseve Cap1 anoratidza ega ega fluorescence emissions emadhayi masere (Dye1-Dye8).Mavara emiseve anoenderana nemavara (b) uye (c).(b) Fluorescence spectra yemadhayi masere (Dye1-Dye8) pa capillary.c Electropherograms yedhayi sere (Dye1-Dye8) pacapillary.Iyo yepamusoro yeDye7-yakanyorwa DNA zvimedu inoratidzwa nemiseve, uye yavo Cap4 base kureba inoratidzwa.
Iyo yakanyanya kugoverwa kweC1-C9 panhongonya masere inoratidzwa muFig.7b, zvichiteerana.Nokuti zvose C1-C9 uye Dye1-Dye8 zviri muwavelength kurongeka, zvikamu zvisere muFig. 7b zvinoratidza fluorescence spectra yeDye1-Dye8 sequentially kubva kuruboshwe kuenda kurudyi.Muchidzidzo ichi, Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, uye Dye8 dzinoonekwa mune magenta, violet, bhuruu, cyan, girinhi, yero, orenji, uye tsvuku zvichiteerana.Cherechedza kuti mavara emiseve ari mumufananidzo 7a anoenderana nemavara edhayi mumufananidzo 7b.Iyo C1-C9 fluorescence intensity kune yega yega spectrum muFigure 7b yakajairwa kuitira kuti mari yavo ienzane.Masere akaenzana efluorescence spectra akawanikwa kubva kuCap1-Cap4.Mumwe anogona kuona zvakajeka kupindirana kwe fluorescence pakati pedhayi 1-dhayi 8.
Sezvinoratidzwa mumufananidzo 7c, kune imwe neimwe capillary, mapfumbamwe-mavara e electrophoregram muFigure 7a yakashandurwa kuita sere-dhayi electropherogram ne-multi-component analysis inoenderana nesere fluorescence spectra muFigure 7b (ona Supplementary Materials for details).Sezvo iyo spectral kupindirana kwefluorescence muFigure 7a isina kuratidzwa muMufananidzo 7c, Dye1-Dye8 inogona kuzivikanwa uye kuverengerwa imwe neimwe panguva imwe neimwe nguva, kunyangwe huwandu hwakasiyana hweDye1-Dye8 fluoresce panguva imwe chete.Izvi hazvigone kuitwa neyakajairwa mavara manomwe-mavara31, asi anogona kuwanikwa neyakagadziridzwa mipfumbamwe-mavara ekuona.Sezvinoratidzwa nemiseve Cap1 muFig. 7c, chete fluorescent emission singlets Dye3 (bhuruu), Dye8 (tsvuku), Dye5 (girini), Dye4 (cyan), Dye2 (yepepuru), Dye1 (magenta), uye Dye6 (Yellow). ) anocherechedzwa munhevedzano inotarisirwa yokutevedzana kwenguva.Nekubuda kwefluorescent kwedhayi 7 (orenji), mukuwedzera kune imwe chete yepamusoro inotaridzwa nemuseve weorenji, mamwe akati wandei mapeaks akaonekwa.Mhedzisiro iyi imhaka yekuti masamples aive nesaizi zviyero, Dye7 yakanyorwa zvidimbu zveDNA zvine hurefu hwakasiyana.Sezvinoratidzwa muMufananidzo 7c, kuCap4 kureba kwehwaro uku i20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 uye 220 mabhesi kureba.
Izvo zvakakosha zvezvipfumbamwe-mavara spectrometer, yakagadziridzwa uchishandisa matrix maviri-layer dichroic magirazi, idiki saizi uye nyore dhizaini.Sezvo huwandu hwemagirazi e decachromatic mukati meiyo adapta inoratidzwa mufig.3c yakasungirirwa zvakananga pamufananidzo we sensor board (ona Fig. S1 uye S2), iyo mipfumbamwe-mavara spectrometer ine zviyero zvakafanana neadapter, kureva 54 × 58 × 8.5 mm.(ukobvu).Iyi ultra-diki saizi maodha maviri kusvika matatu ehukuru madiki pane akajairwa spectrometers anoshandisa gratings kana prisms.Pamusoro pezvo, sezvo iyo mipfumbamwe-mavara spectrometer yakagadziridzwa zvekuti mwenje unorova pamusoro peiyo sensor yemufananidzo perpendicularly, nzvimbo inogona kugoverwa nyore kune mapfumbamwe-mavara spectrometer mumasisitimu akadai semamicroscopes, flow cytometers, kana analyzer.Capillary grating electrophoresis analyzer kune yakatokura miniaturization yehurongwa.Panguva imwecheteyo, saizi yegumi dichroic magirazi uye bandpass mafirita anoshandiswa mune mapfumbamwe-mavara spectrometer ndeye 10 × 1.9 × 0.5 mm kana 15 × 1.9 × 0.5 mm.Nekudaro, anopfuura zana akadai madiki dichroic magirazi uye bandpass mafirita, zvichiteerana, anogona kutemwa kubva kune dichroic girazi uye 60 mm2 bandpass sefa, zvichiteerana.Naizvozvo, ruzhinji rwemagirazi e decachromatic anogona kugadzirwa nemutengo wakaderera.
Chimwe chimiro chezvipfumbamwe-mavara spectrometer ndiyo yakanakisa spectral maitiro.Kunyanya, inobvumira kuwanikwa kwemifananidzo ye spectral ye snapshots, ndiko kuti, panguva imwe chete yekutora mifananidzo ine ruzivo rwe spectral.Pamufananidzo wega wega, chiratidziro chinoenderera chakawanikwa newavelength kubva pa520 kusvika 700 nm uye resolution ye20 nm.Nemamwe manzwi, kusimba kwemavara mapfumbamwe kunoonekwa pamufananidzo wega wega, kureva mapfumbamwe 20 nm mabhendi anopatsanura zvakaenzana kureba kwewavelength kubva pa520 kusvika 700 nm.Nekushandura spectral maitiro eiyo dichroic girazi uye bandpass sefa, iyo wavelength huwandu hwemapfumbamwe mabhendi uye hupamhi hwebhendi rega rega inogona kugadziriswa.Mapfumbamwe emavara ekuona anogona kushandiswa kwete chete kune fluorescence kuyerwa ne spectral imaging (sezvinotsanangurwa mumushumo uyu), asiwo kune mamwe akawanda akajairika maapplication uchishandisa spectral imaging.Kunyange zvazvo hyperspectral imaging inogona kuona mazana emavara, zvakaonekwa kuti kunyange nekuderera kukuru kwenhamba yemavara anoonekwa, zvinhu zvakawanda mumunda wekuona zvinogona kuonekwa nekururama kwakakwana kwezvikumbiro zvakawanda38,39,40.Nekuti kugadzirisa kwenzvimbo, kutaridzika kwekutarisa, uye kugadzirisa kwechinguva kune tradeoff mukufungidzira kwekuona, kuderedza huwandu hwemavara kunogona kuvandudza kugadziriswa kwenzvimbo uye kugadzirisa kwechinguva.Inogona zvakare kushandisa ma spectrometer akareruka senge akagadzirwa muchidzidzo ichi uyezve kuderedza huwandu hwemakomputa.
Muchidzidzo ichi, madhayi masere akaverengerwa panguva imwe chete nekupatsanurwa kwe spectral yavo inoputika fluorescence spectra zvichibva pakuonekwa kwemavara mapfumbamwe.Anosvika madhayi mapfumbamwe anogona kuverengerwa panguva imwe chete, achigarisana munguva nenzvimbo.Mukana wakakosha weiyo mipfumbamwe-mavara spectrometer ndeye yakakwira inopenya flux uye yakakura aperture (1 × 7 mm).Iyo decane mirror array ine yakanyanya kutapurirana inosvika 92% yechiedza kubva pakavhurika mune imwe neimwe yemapfumbamwe ewavelength.Kubudirira kwekushandisa chiitiko mwenje muwavelength kubva pa520 kusvika 700 nm ingangoita 100%.Muhupamhi hwakadai hwewavelengths, hapana diffraction grating inogona kupa yakanyanya kunaka kwekushandisa.Kunyange kana kupatsanurwa kwegetsi kwegetsi kuchipfuura 90% pane imwe wavelength, sezvo musiyano uripo pakati peiyo wavelength uye imwe wavelength unowedzera, kusiyanisa kushanda kune imwe wavelength kunoderera41.Iyo aperture wide perpendicular to the direction of the plane in Fig. 2c inogona kuwedzerwa kubva pa7 mm kusvika pahupamhi hwechifananidzo chemufananidzo, zvakadai sechiitiko chechimiro chemufananidzo chinoshandiswa muchidzidzo ichi, nekugadzirisa zvishoma decamer array.
Iyo mipfumbamwe-mavara spectrometer inogona kushandiswa kwete chete kune capillary electrophoresis, sezvakaratidzwa muchidzidzo ichi, asiwo kune zvimwe zvinangwa zvakasiyana.Semuenzaniso, sezvakaratidzwa mumufananidzo uri pasi apa, spectrometer ine mavara mapfumbamwe inogona kuiswa kune fluorescence microscope.Iyo ndege yemuenzaniso inoratidzwa pane sensor yemifananidzo yepfumbamwe-ruvara spectrometer kuburikidza ne10x chinangwa.Iyo yekuona kureba pakati pechinangwa lens uye sensor yemufananidzo ndeye 200 mm, nepo nhambwe yekuona pakati pechiitiko cheiyo mapfumbamwe-ruvara spectrometer uye mufananidzo sensor inongova 12 mm.Nokudaro, chifananidzo chacho chakachekwa kusvika pakukura kweaperture (1 × 7 mm) mundege yezviitiko uye yakakamurwa kuva mipfumbamwe mifananidzo yemavara.Ndiko kuti, mufananidzo we spectral we pfumbamwe-mavara snapshot inogona kutorwa pane 0.1 × 0.7 mm nzvimbo mumuenzaniso ndege.Mukuwedzera, zvinokwanisika kuwana mapfumbamwe-mavara spectral mufananidzo wenzvimbo yakakura pane imwe ndege yemuenzaniso nekuongorora sampuli maererano nechinangwa mugwara rakadzika muFig. 2c.
Iyo decachromatic mirror array components, zvinoti M1-M9 uye BP, dzakagadzirwa neAsahi Spectra Co., Ltd. vachishandisa nzira dzakajairwa dzekunaya kwemvura.Multilayer dielectric zvinhu zvakaiswa mumwe nemumwe pamapureti gumi equartz 60 × 60 mm muhukuru uye 0.5 mm gobvu, achizadzisa zvinotevera zvinodiwa: M1: IA = 45°, R ≥ 90% pa520–590 nm, Tave ≥ 90% pa610– 610 nm.700 nm, M2: IA = 45 °, R ≥ 90% pa520-530 nm, Tave ≥ 90% pa550-600 nm, M3: IA = 45 °, R ≥ 90% pa540-550 nm, Tave 0 % pa570-600 nm, M4: IA = 45 °, R ≥ 90% pa560-570 nm, Tave ≥ 90% pa590-600 nm, M5: IA = 45 °, R ≥ 98% pa580-600 nm , R ≥ 98% pa 680-700 nm, M6: IA = 45 °, Tave ≥ 90% pa 600-610 nm, R ≥ 90% pa 630-700 nm, M7: IA = 45 °, R ≥ 90% 620-630 nm, Taw ≥ 90% pa 650-700 nm, M8: IA = 45 °, R ≥ 90% pa 640-650 nm, Taw ≥ 90% pa 670-700 nm, M9: IA = 45 ° ≥ 90% pa 650-670 nm, Tave ≥ 90% pa 690-700 nm, BP: IA = 0 °, T ≤ 0.01% pa 505 nm, Tave ≥ 95% pa 530-690 nm pa 530 nm T0≥ pa -690 nm uye T ≤ 1% pa725-750 nm, apo IA, T, Tave, uye R ndiyo angle yezviitiko, transmittance, avhareji transmittance, uye unpolarized chiedza kuratidza.
Chiedza chichena (C0) chine urefu hwemasaisai e400–750 nm inoburitswa nemwenje we LED sosi (AS 3000, AS ONE CORPORATION) yakasanganiswa uye yakaitika yakatwasuka paDP yemhando yezvionioni zvedichroic.Iyo chena mwenje spectrum ye LED inoratidzwa muSupplementary Figure S3.Isa acrylic tank (zviyero 150 × 150 × 30 mm) zvakananga pamberi peiyo decamera mirror array, yakatarisana nePSU.Hutsi hwakagadzirwa pakanyudzwa aizi yakaoma mumvura yakabva yadirwa muacrylic tank kuti itarise hova pfumbamwe dzeC1-C9 dzakatsemurwa dzichibva murutanho rwemagirazi edecachromatic.
Neimwe nzira, iyo collimated white light (C0) inopfuudzwa nepasefa isati yapinda muDP.Masefa aya aive mafirita asina kwaakarerekera ane density ye0.6.Wobva washandisa sefa ine mota (FW212C, FW212C, Thorlabs).Pakupedzisira, dzorera ND sefa.Iwo bandwidths emapfumbamwe ebhendi mafirita anoenderana neC9, C8, C7, C6, C5, C4, C3, C2 uye C1, zvichiteerana.Sero re quartz rine zviyero zvemukati zve 40 (kureba kwemaziso) x 42.5 (kureba) x 10 mm (upamhi) yakaiswa pamberi pezvioni zve decochromatic, zvakatarisana neBP.Hutsi hunozopihwa nemuchubhu kupinda musero requartz kuchengetedza hutsi hwehutsi musero requartz kuti utarise hova pfumbamwe dzeC1-C9 dzakapatsanurwa dzinobuda kubva mugirazi re decachromatic array.
Vhidhiyo yeiyo mipfumbamwe-mavara akapatsanurwa mwenje rukova runobva kune akatevedzana e decanic magirazi akatorwa mu-time-lapse mode pa iPhone XS.Bata mifananidzo yechiitiko pa1 fps uye gadzira iyo mifananidzo kugadzira vhidhiyo pa30 fps (yevhidhiyo sarudzo 1) kana 24 fps (yemavhidhiyo esarudzo 2 uye 3).
Isa 50 µm gobvu ndiro yesimbi isina tsvina (ine maburi mana 50 µm dhayamita pa 1 mm intervals) padiffusion plate.Chiedza chine wavelength ye400-750 nm chinovhenekerwa pandiro ye diffuser, inowanikwa nekupfuura mwenje kubva pamwenje wehalogen kuburikidza nepfupi yekufambisa sefa ine cutoff wavelength ye700 nm.Chiedza chechiedza chinoratidzwa muSupplementary Figure S4.Neimwe nzira, mwenje unopfuurawo neimwe yegumi nm bandpass mafirita akatarisana ne530, 550, 570, 590, 610, 630, 650, 670 uye 690 nm uye inorova diffuser ndiro.Nekuda kweizvozvo, mapoinzi mana emwaranzi ane dhayamita ye φ50 μm uye akasiyana wavelengths akaumbwa pandiro yesimbi isina tsvina yakatarisana nediffuser plate.
A mana-capillary array ane mana lenzi akaiswa pane mapfumbamwe-mavara spectrometer sezvakaratidzwa muFigure 1 uye 2. C1 uye C2.Iwo mana capillaries uye mana lenzi aive akafanana neapfuura zvidzidzo31,34.Laser beam ine wavelength ye505 nm uye simba re 15 mW inovhenekerwa panguva imwe chete uye zvakaenzana kubva padivi kuenda kunzvimbo dzekubuda kwema capillaries mana.Iyo fluorescence inoburitswa nenzvimbo yega yega inobuda inobatanidzwa nelenzi inoenderana uye yakapatsanurwa kuita mipfumbamwe yemavara hova nemhando yezvionioni zve decachromatic.Mhedzisiro 36 hova dzakazoiswa zvakananga muCMOS mufananidzo sensor (C11440-52U, Hamamatsu Photonics K · K.), uye mifananidzo yavo yakarekodhwa panguva imwe chete.
ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Applied Biosystems), 4 µl GeneScan™ 600 LIZ™ dhayi yakasanganiswa yecapillary yega yega nekusanganisa 1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), 1 µl musanganiswa saizi standard.v2.0 (Thermo Fisher Scientific) uye 14 µl yemvura.PowerPlex® 6C Matrix Standard ine zvimedu zveDNA zvitanhatu zvakanyorwa nedhayi nhanhatu: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, uye WEN, maererano nehupamhi hwemasaisai.Hwaro hurefu hwezvimedu zveDNA izvi hahuziviswe, asi kutevedzana kwehwaro hwezvimedu zveDNA zvakanyorwa neWEN, CXR-6C, TMR-6C, JOE-6C, FL-6C uye TOM-6C inozivikanwa.Musanganiswa uri muABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit ine DNA fragment yakanyorwa nedhayi yedR6G.Hurefu hwemabhesi ezvimedu zveDNA hazvina kuburitswa.GeneScan™ 600 LIZ™ Dye Size Standard v2.0 inosanganisira makumi matatu nematanhatu LIZ-yakanyorwa zvidimbu zveDNA.Hwaro hurefu hwezvikamu zveDNA izvi 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 40, 301, 30 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 uye 600 base.Iwo masampuli akaiswa denatured pa 94 ° C kwemaminetsi matatu, ndokuzonyoroveswa pachando kwemaminitsi mashanu.Samples dzakabayiwa mucapillary yega yega pa26 V/cm kwe9 s uye yakapatsanurwa mune yega yega capillary yakazadzwa nePOP-7™ polymer solution (Thermo Fisher Scientific) ine hurefu hunoshanda hwe36 cm uye voltage ye181 V/cm uye an kona ye60 °.KUBVA.
Yese data yakawanikwa kana kuongororwa mukati mechidzidzo ichi inosanganisirwa mune ino yakabudiswa chinyorwa uye neruzivo rwayo rwekuwedzera.Mamwe data anoenderana nechidzidzo ichi anowanikwa kubva kune vakasiyana vanyori kana paine chikumbiro chine musoro.
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