zitsulo zosapanga dzimbiri 321 8 * 1.2 chubu chophimbidwa chosinthira kutentha

Machubu a Capillary

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Chowunikira chowoneka bwino kwambiri (54 × 58 × 8.5 mm) ndi kabowo kakang'ono (1 × 7 mm) chamitundu isanu ndi inayi chinapangidwa, "chogawikana pawiri" ndi mndandanda wa magalasi khumi a dichroic, omwe amagwiritsidwa ntchito pompopompo kujambula.The chochitika kuwala flux ndi mtanda gawo ang'onoang'ono kuposa kabowo kukula amagawidwa mosalekeza Mzere 20 nm lonse ndi naini mtundu fluxes ndi wavelengths chapakati 530, 550, 570, 590, 610, 630, 650, 670 ndi 690 nm.Zithunzi za mitsinje isanu ndi inayi zimayesedwa nthawi imodzi ndi sensor yazithunzi.Mosiyana ndi magalasi ochiritsira a dichroic, gulu lopangidwa ndi galasi la dichroic lili ndi kasinthidwe kaŵirikaŵiri, zomwe sizimangowonjezera kuchuluka kwa mitundu yomwe ingayesedwe panthawi imodzi, komanso kumapangitsanso kusintha kwazithunzi pamtundu uliwonse wamtundu.Ma spectrometer opangidwa ndi mitundu isanu ndi inayi amagwiritsidwa ntchito pa ma electrophoresis a capillary anayi.Kusanthula kwakanthawi kofanana kwa mitundu isanu ndi itatu yomwe imasuntha nthawi imodzi mu capillary iliyonse pogwiritsa ntchito fluorescence yamitundu isanu ndi inayi.Popeza spectrometer yamitundu isanu ndi inayi sikuti ndi yaying'ono komanso yotsika mtengo, komanso imakhala ndi kuwala kowoneka bwino komanso mawonekedwe owoneka bwino pamawonekedwe ambiri owonera, imatha kugwiritsidwa ntchito kwambiri m'magawo osiyanasiyana.
Kujambula kwa hyperspectral ndi multispectral kwakhala gawo lofunika kwambiri la zakuthambo2, kuyang'ana kutali kwa Earth observation3,4, kuyang'anira khalidwe la chakudya ndi madzi5,6, kusungirako zojambulajambula ndi zakale7, forensics8, surgery9, biomedical analysis and diagnostics10,11 etc. Munda 1 Ukadaulo wofunikira kwambiri , 12, 13.Njira zoyezera kuchuluka kwa kuwala komwe kumatulutsa ndi gawo lililonse lazomwe zimawonekera zimagawidwa mu (1) kusanthula mfundo ("tsache")14,15, (2) kusanthula kwa mzere ("panicle")16,17,18 , (3) kutalika kwa mafunde19,20,21 ndi (4) zithunzi22,23,24,25.Pankhani ya njira zonsezi, kusintha kwa malo, mawonekedwe a spectral ndi kusintha kwakanthawi kumakhala ndi mgwirizano wamalonda9,10,12,26.Kuphatikiza apo, kutulutsa kowala kumakhudza kwambiri kukhudzika, mwachitsanzo, chiŵerengero cha ma signal-to-noise mu spectral imaging26.Kuwala kowala, ndiko kuti, mphamvu yogwiritsira ntchito kuwala, imagwirizana mwachindunji ndi chiŵerengero cha kuchuluka kwenikweni kwa kuwala kwa malo aliwonse owala pa nthawi ya unit mpaka kuchuluka kwa kuwala kwa msinkhu woyezedwa wa wavelength.Gulu (4) ndi njira yoyenera pamene mphamvu kapena sipekitiramu ya kuwala kotulutsidwa ndi mfundo iliyonse yotulutsa imasintha ndi nthawi kapena pamene malo a malo aliwonse otulutsa asintha ndi nthawi chifukwa kuwala kotulutsidwa ndi mfundo zonse zotulutsa kumayesedwa nthawi imodzi.24.
Njira zambiri zomwe zili pamwambazi zimaphatikizidwa ndi ma spectrometer akulu, ovuta komanso / kapena okwera mtengo pogwiritsa ntchito ma grating 18 kapena 14, 16, 22, 23 ma prisms a makalasi (1), (2) ndi (4) kapena 20, 21 zosefera zamadzimadzi. .Zosefera za Crystalline tunable (LCTF)25 kapena zosefera za acousto-optic tunable (AOTF)19 za gulu (3).Mosiyana ndi zimenezi, gulu (4) ma spectrometer a magalasi ambiri ndi ochepa komanso otsika mtengo chifukwa cha kuphweka kwawo27,28,29,30.Kuonjezera apo, ali ndi kuwala kowala kwambiri chifukwa kuwala komwe kumagawidwa ndi galasi lililonse la dichroic (ndiko kuti, kuwala kowonetsera ndi kuwonetseredwa kwa kuwala kwa zochitika pa galasi lililonse la dichroic) kumagwiritsidwa ntchito mokwanira komanso mosalekeza.Komabe, kuchuluka kwa mafunde a kutalika kwa mafunde (ie mitundu) yomwe imayenera kuyezedwa nthawi imodzi ndi pafupifupi anayi.
Kujambula kwapadera kozikidwa pa kufufuza kwa fluorescence kumagwiritsidwa ntchito kawirikawiri pofufuza ma multiplex mu kufufuza kwa biomedical ndi diagnostics 10, 13.Mu multiplexing, popeza ma analyte angapo (mwachitsanzo, DNA yeniyeni kapena mapuloteni) amalembedwa ndi utoto wosiyanasiyana wa fulorosenti, analyte aliwonse omwe amapezeka pamtundu uliwonse wa kawonedwe amayesedwa pogwiritsa ntchito kusanthula kwamagulu ambiri.32 imaphwanya mawonekedwe a fluorescence omwe amaperekedwa ndi malo aliwonse otulutsa.Panthawi imeneyi, utoto wosiyanasiyana, uliwonse umatulutsa fulorosenti yosiyana, ukhoza kusungunuka, ndiko kuti, kukhala pamodzi mumlengalenga ndi nthawi.Pakadali pano, kuchuluka kwa utoto komwe kumatha kusangalatsidwa ndi mtengo umodzi wa laser ndi eyiti33.Malire apamwambawa samatsimikiziridwa ndi mawonekedwe a spectral (ie, kuchuluka kwa mitundu), koma ndi m'lifupi mwa sipekitiramu ya fluorescence (≥50 nm) ndi kuchuluka kwa utoto wa Stokes shift (≤200 nm) pa FRET (pogwiritsa ntchito FRET)10 .Komabe, kuchuluka kwa mitundu kuyenera kukhala kokulirapo kapena kofanana ndi kuchuluka kwa utoto kuti athetse kuphatikizika kwamitundu yosakanikirana31,32.Choncho, m'pofunika kuwonjezera chiwerengero cha mitundu yoyezera nthawi imodzi kufika eyiti kapena kuposerapo.
Posachedwapa, ultra-compact heptachroic spectrometer (yogwiritsa ntchito mitundu yosiyanasiyana ya magalasi a heptychroic ndi sensa ya zithunzi poyeza ma fluorescent fluxes) yapangidwa.Ma spectrometer ndi madongosolo awiri kapena atatu a kukula kocheperako kuposa ma spectrometer wamba pogwiritsa ntchito ma grating kapena ma prisms34,35.Komabe, nkovuta kuyika magalasi oposa asanu ndi awiri a dichroic mu spectrometer ndipo nthawi imodzi amayesa mitundu yoposa isanu ndi iwiri36,37.Ndi kuwonjezeka kwa chiwerengero cha magalasi a dichroic, kusiyana kwakukulu kwa kutalika kwa njira za kuwala kwa dichroic light fluxes kumawonjezeka, ndipo zimakhala zovuta kusonyeza kuwala konse pa ndege imodzi yomvera.Kutalika kwa njira yayitali kwambiri ya kuwala kwa kuwala kumawonjezekanso, motero m'lifupi mwake kabowo ka spectrometer (ie kukula kwake kwa kuwala koyesedwa ndi spectrometer) kumachepa.
Poyankha mavuto omwe ali pamwambawa, chowonera chamitundu isanu ndi inayi chokhala ndi magalasi osanjikiza awiri a "dichroic" decachromatic ndi sensa ya zithunzi zowonera pompopompo [gulu (4)].Poyerekeza ndi ma spectrometer am'mbuyomu, ma spectrometer opangidwa ali ndi kusiyana kocheperako panjira yotalikirapo komanso njira yaying'ono yocheperako.Agwiritsidwa ntchito pa ma electrophoresis a capillary anayi kuti azindikire fluorescence yamitundu isanu ndi inayi yopangidwa ndi laser komanso kuwerengera kusuntha kwamitundu isanu ndi itatu mu capillary iliyonse.Popeza kuti spectrometer yopangidwa sikhala yocheperako komanso yotsika mtengo, komanso imakhala ndi kuwala kowala kwambiri komanso mawonekedwe owoneka bwino pamawonekedwe ambiri owonera, imatha kugwiritsidwa ntchito kwambiri m'magawo osiyanasiyana.
Chowonera chamitundu isanu ndi inayi chikuwonetsedwa mkuyu.1 a.Mapangidwe ake amatsatira a ultra-ang'ono ang'onoang'ono amitundu isanu ndi iwiri spectrometer 31. Amakhala ndi magalasi asanu ndi anayi a dichroic okonzedwa mozungulira pamtunda wa 45 ° kumanja, ndipo chithunzithunzi cha chithunzi (S) chili pamwamba pa magalasi asanu ndi anayi a dichroic.Kuwala komwe kumalowa kuchokera pansi (C0) kumagawidwa ndi magalasi asanu ndi anayi a dichroic kukhala maulendo asanu ndi anayi akuyenda mmwamba (C1, C2, C3, C4, C5, C6, C7, C8 ndi C9).Mitsinje yonse isanu ndi inayi imadyetsedwa mwachindunji ku sensa ya zithunzi ndipo imapezeka nthawi imodzi.Mu phunziro ili, C1, C2, C3, C4, C5, C6, C7, C8, ndi C9 ali mu dongosolo la kutalika kwa mafunde ndipo amaimiridwa ndi magenta, violet, blue, cyan, green, yellow, lalanje, red-orange, ndi red, motero.Ngakhale kuti mitundu imeneyi imagwiritsidwa ntchito m’chikalatachi, monga mmene chithunzi 3 chasonyezera, chifukwa imasiyana ndi mitundu yeniyeni imene maso a munthu amaona.
Zojambula zamakina zama spectrometer odziwika bwino komanso atsopano amitundu isanu ndi inayi.(a) Magalasi owoneka bwino amitundu isanu ndi inayi okhala ndi mizere isanu ndi inayi.(b) Chowonera chatsopano chamitundu isanu ndi inayi chokhala ndi magalasi awiri osanjikizana a dichroic.Chochitika chowunikira chowunikira C0 chimagawidwa m'mitundu isanu ndi inayi yowunikira C1-C9 ndikuzindikiridwa ndi sensa ya chithunzi S.
Chojambula chatsopano chamitundu isanu ndi inayi chimakhala ndi galasi la dichroic lamitundu iwiri ndi sensa ya chithunzi, monga momwe tawonetsera mkuyu 1b.M'munsi mwake, magalasi asanu a dichroic amapendekeka 45 ° kumanja, olumikizidwa kumanja kuchokera pakati pa gulu la decamers.Pamwamba, magalasi asanu owonjezera a dichroic amapendekeka 45 ° kumanzere ndipo amakhala kuchokera pakati kupita kumanzere.Kalilore wa kumanzere wa dichroic wosanjikiza wa m'munsi ndi galasi lamanja la dichroic lapamwamba limadutsana.Kuwala kochitika (C0) kumagawidwa kuchokera pansi kukhala ma chromatic fluxes anayi (C1-C4) ndi magalasi asanu a dichroic kumanja ndi ma chromatic fluxes asanu (C5-C4) ndi magalasi asanu a dichroic kumanzere C9).Monga ma spectrometer wamba amitundu isanu ndi inayi, mitsinje yonse isanu ndi inayi imabayidwa mwachindunji mu sensa ya zithunzi (S) ndikuzindikiridwa nthawi imodzi.Poyerekeza Zithunzi 1a ndi 1b, munthu akhoza kuona kuti pamtundu wa spectrometer yatsopano yamitundu isanu ndi inayi, kusiyana kwakukulu komanso kutalika kwa njira yotalikirapo ya mitundu isanu ndi inayi yamitundu yosiyanasiyana ndi theka.
Kumanga mwatsatanetsatane kwa galasi laling'ono laling'ono laling'ono la dichroic 29 mm (m'lifupi) × 31 mm (kuya) × 6 mm (kutalika) likuwonetsedwa mu Chithunzi 2. Gulu la galasi la dichroic lili ndi magalasi asanu a dichroic kumanja. (M1-M5) ndi magalasi asanu a dichroic kumanzere (M6-M9 ndi M5 wina), galasi lililonse la dichroic limayikidwa muzitsulo za aluminiyamu zapamwamba.Magalasi onse a dichroic amasunthidwa kuti alipirire kusamutsidwa kofananira chifukwa cha kusinthika kwakuyenda kudzera pagalasi.Pansi pa M1, fyuluta ya band-pass (BP) imakhazikika.M1 ndi BP miyeso ndi 10mm (mbali yaitali) x 1.9mm (mbali yaifupi) x 0.5mm (kukhuthala).Miyeso ya magalasi otsala a dichroic ndi 15 mm × 1.9 mm × 0.5 mm.Kutalika kwa matrix pakati pa M1 ndi M2 ndi 1.7 mm, pamene matrix a magalasi ena a dichroic ndi 1.6 mm.Pa mkuyu.2c imaphatikiza kuwala kochitika C0 ndi kuwala kowala kwamitundu isanu ndi inayi C1-C9, olekanitsidwa ndi magalasi a de-chamber.
Kupanga matrix amitundu iwiri ya dichroic.(a) Kawonedwe kawonekedwe ndi (b) mawonekedwe ophatikizika a magalasi awiri osanjikiza a dichroic array (miyeso 29 mm x 31 mm x 6 mm).Zimapangidwa ndi magalasi asanu a dichroic (M1-M5) omwe ali pansi pamunsi, magalasi asanu a dichroic (M6-M9 ndi M5 wina) omwe ali pamtunda wapamwamba, ndi fyuluta ya bandpass (BP) yomwe ili pansi pa M1.(c) Mawonedwe amtundu wodutsa molunjika, ndi C0 ndi C1-C9 zikuphatikizana.
M'lifupi kabowo mu njira yopingasa, anasonyeza ndi m'lifupi C0 mu mkuyu 2, c, ndi 1 mm, ndi mbali perpendicular ndege ya mkuyu. -7 mm.Ndiko kuti, spectrometer yatsopano yamitundu isanu ndi inayi ili ndi kukula kwakukulu kwa 1 mm × 7 mm.Njira yowunikira ya C4 ndiyo yayitali kwambiri pakati pa C1-C9, ndipo njira yowonera ya C4 mkati mwa galasi la dichroic, chifukwa cha kukula kwake kochepa kwambiri (29 mm × 31 mm × 6 mm), ndi 12 mm.Pa nthawi yomweyo, kuwala njira kutalika C5 ndi lalifupi pakati pa C1-C9, ndi kuwala njira kutalika C5 ndi 5.7mm.Choncho, kusiyana kwakukulu mu kutalika kwa njira ya kuwala ndi 6.3 mm.Kutalika kwa njira ya pamwambayi kumakonzedweratu chifukwa cha kutalika kwa njira ya kuwala kwa kuwala kwa M1-M9 ndi BP (kuchokera ku quartz).
Mawonekedwe owoneka bwino a М1−М9 ndi VR amawerengedwa kotero kuti mafunde С1, С2, С3, С4, С5, С6, С7, С8 ndi С9 ali pamtunda wa 520-540, 540-560, 560-8580, 560-8580, 560-8580, 580-580 -600 , 600-620, 620-640, 640-660, 660-680, ndi 680-700 nm, motero.
Chithunzi cha matrix opangidwa ndi magalasi a decachromatic akuwonetsedwa mkuyu 3a.M1-M9 ndi BP zimamangiriridwa kumalo otsetsereka a 45 ° ndi ndege yopingasa ya chithandizo cha aluminiyumu, motero, pamene M1 ndi BP zimabisika kumbuyo kwa chithunzicho.
Kupanga magalasi angapo a decan ndi mawonetsero ake.(a) Magalasi angapo opangidwa ndi decachromatic.(b) Chithunzi chogawanika cha 1 mm × 7 mm chamitundu isanu ndi inayi chojambulidwa papepala loyikidwa kutsogolo kwa magalasi osasunthika komanso owala kumbuyo ndi kuwala koyera.(c) Magalasi angapo a decochromatic owala ndi kuwala koyera kuchokera kumbuyo.(d) Mitsinje yogawanika yamitundu isanu ndi inayi yochokera pagalasi la decane, imawonedwa poyika chitini cha acrylic chodzaza utsi kutsogolo kwa galasi la decane pa c ndikupangitsa chipindacho mdima.
Mawonekedwe opatsirana oyezedwa a M1-M9 C0 pa ngodya ya 45 ° ndi kufalikira kwa BP C0 pakona ya zochitika za 0 ° akuwonetsedwa mu Fig.4 a.Mawonekedwe opatsirana a C1-C9 okhudzana ndi C0 akuwonetsedwa muzithunzi.4b .Mawonekedwe awa adawerengedwa kuchokera ku spectra mu Mkuyu.4a mogwirizana ndi njira ya kuwala C1-C9 mu Mkuyu 4a.1b ndi 2c.Mwachitsanzo, 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)], pamene TS(X) ndi [ 1 − TS(X)] ndi mawonekedwe a X, motsatana.Monga momwe tawonetsera mu Chithunzi 4b, ma bandwidth (bandwidth ≥50%) a C1, C2, C3, C4, C5, C6, C7, C8 ndi C9 ndi 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 ndi 682-699 nm.Zotsatirazi zimagwirizana ndi magulu otukuka.Kuphatikiza apo, kugwiritsa ntchito bwino kwa kuwala kwa C0 ndikokwera, ndiye kuti, pafupifupi pafupifupi C1-C9 kuwala kwapakatikati ndi 92%.
Mawonekedwe opatsirana a galasi la dichroic ndi kugawanika kwa mitundu isanu ndi inayi.(a) Kuyesedwa kwa kufalikira kwa M1-M9 pa zochitika za 45 ° ndi BP pa 0 °.(b) Mawonekedwe opatsira a C1–C9 okhudzana ndi C0 owerengedwa kuchokera ku (a).
Pa mkuyu.3c, magalasi amtundu wa dichroic amapezeka molunjika, kotero kuti mbali yake yakumanja mu Chithunzi 3a ndi mbali ya pamwamba ndipo mtengo woyera wa collimated LED (C0) umabwereranso.Mitundu yambiri ya magalasi a decachromatic yomwe ikuwonetsedwa mu Chithunzi 3a imayikidwa mu 54 mm (kutalika) × 58 mm (kuya) × 8.5 mm (kukhuthala) adaputala.Pa mkuyu.3d, kuwonjezera pa dziko lomwe likuwonetsedwa mkuyu.3c, thanki ya acrylic yodzaza utsi inayikidwa patsogolo pa magalasi angapo a decochromatic, ndi magetsi omwe ali m'chipindamo azimitsidwa.Zotsatira zake, mitsinje isanu ndi inayi ya dichroic ikuwoneka mu thanki, yochokera ku magalasi angapo a decachromatic.Mtsinje uliwonse wogawanika uli ndi gawo lamtanda lamakona anayi okhala ndi miyeso ya 1 × 7 mm, yomwe imagwirizana ndi kukula kwa kabowo ka spectrometer yatsopano yamitundu isanu ndi inayi.Pachithunzi 3b, pepala layikidwa kutsogolo kwa magalasi amtundu wa dichroic mu Chithunzi 3c, ndipo chithunzi cha 1 x 7 mm cha mitsinje isanu ndi inayi ya dichroic yomwe ikuwonetsedwa papepala ikuwoneka kuchokera kumbali ya kayendedwe ka pepala.mitsinje.Mitsinje isanu ndi inayi yolekanitsa mitundu mu mkuyu.3b ndi d ndi C4, C3, C2, C1, C5, C6, C7, C8 ndi C9 kuchokera pamwamba mpaka pansi, zomwe zimawonekanso muzithunzi 1 ndi 2. 1b ndi 2c.Amawonedwa mumitundu yogwirizana ndi kutalika kwake.Chifukwa cha kuwala kochepa koyera kwa LED (onani Supplementary Fig. S3) ndi kutengeka kwa kamera yamtundu yomwe imagwiritsidwa ntchito kujambula C9 (682-699 nm) mumkuyu. Zina zogawanika zimayenda ndi zofooka.Momwemonso, C9 inkawoneka bwino m'maso.Panthawiyi, C2 (mtsinje wachiwiri kuchokera pamwamba) umawoneka wobiriwira mu Chithunzi 3, koma umawoneka wachikasu kwambiri m'maso.
Kusintha kuchokera ku Chithunzi 3c kupita ku d kukuwonetsedwa mu Supplementary Video 1. Nthawi yomweyo kuwala koyera kuchokera ku LED kudutsa mu galasi la decachromatic, kumagawanika panthawi imodzi mu mitsinje isanu ndi inayi.Pamapeto pake, utsi wa mtsukowo unatha pang'onopang'ono kuchokera pamwamba mpaka pansi, kotero kuti ma ufa asanu ndi anayi amtunduwo adasowa kuchokera pamwamba mpaka pansi.Mosiyana ndi izi, mu Supplementary Video 2, pamene kutalika kwa mawonekedwe a kuwala kwa magalasi a decachromatic kunasinthidwa kuchoka patali kupita kufupi mwa dongosolo la 690, 671, 650, 632, 610, 589, 568, 550 ndi 532 nm. ., Mitsinje yokhayo yogawanika ya mitsinje isanu ndi inayi yogawanika mu dongosolo la C9, C8, C7, C6, C5, C4, C3, C2, ndi C1 ikuwonetsedwa.Chitsime cha acrylic chimasinthidwa ndi dziwe la quartz, ndipo ma flakes a shunted flow iliyonse amatha kuwonedwa momveka bwino kuchokera kumayendedwe otsetsereka.Kuphatikiza apo, vidiyo yaying'ono 3 imasinthidwa kotero kuti gawo la kusintha kwa mawonekedwe a kanema kakang'ono 2 libwerezedwanso.Uwu ndiye mawonekedwe omveka bwino a mawonekedwe a magalasi a decochromatic.
Zotsatira zomwe zili pamwambazi zikuwonetsa kuti magalasi opangidwa ndi decachromatic kapena spectrometer yatsopano yamitundu isanu ndi inayi imagwira ntchito monga momwe amafunira.Chowonera chatsopano chamitundu isanu ndi inayi chimapangidwa ndikuyika magalasi angapo a decachromatic okhala ndi ma adapter pa board sensor.
Kuwala kowala kokhala ndi kutalika kwa kutalika kwa 400 mpaka 750 nm, komwe kumapangidwa ndi mfundo zinayi za radiation φ50 μm, yomwe ili pamtunda wa 1 mm kumbali ya ndege ya mkuyu. magalasi anayi φ1 mm ndi kutalika kwa 1.4 mm ndi phula la 1 mm.Mitsinje inayi yosakanikirana (inayi C0) ikuchitika pa DP ya spectrometer yatsopano yamitundu isanu ndi inayi, yotalikirana ndi 1 mm.Magalasi angapo a dichroic amagawa mtsinje uliwonse (C0) kukhala mitsinje isanu ndi inayi (C1-C9).Mitsinje 36 (ma seti anayi a C1-C9) kenako amabayidwa mwachindunji mu sensa ya zithunzi za CMOS (S) yolumikizidwa mwachindunji ndi magalasi angapo a dichroic.Chotsatira chake, monga momwe tawonetsera mkuyu 5a, chifukwa cha kusiyana kochepa kwambiri kwa kuwala kwa njira ndi njira yaying'ono yowonongeka, zithunzi za mitsinje yonse ya 36 zinadziwika panthawi imodzi komanso momveka bwino ndi kukula kwake.Malinga ndi mawonekedwe akumunsi (onani Supplementary Figure S4), mphamvu yazithunzi zamagulu anayi C1, C2 ndi C3 ndizochepa.Zithunzi makumi atatu ndi zisanu ndi chimodzi zinali 0.57 ± 0.05 mm kukula (kutanthauza ± SD).Chifukwa chake, kukula kwachithunziko kunali pafupifupi 11.4.Kutalikirana koyima pakati pa zithunzi kumakhala pafupifupi 1 mm (mipata yofanana ndi gulu la lens) ndipo mipata yopingasa imakhala pafupifupi 1.6 mm (mipata yofanana ndi mindandanda yagalasi ya dichroic).Chifukwa kukula kwa chithunzicho ndi kochepa kwambiri kusiyana ndi mtunda wapakati pa zithunzi, chithunzi chilichonse chikhoza kuyesedwa paokha (ndi low crosstalk).Pakalipano, zithunzi za mitsinje makumi awiri ndi zisanu ndi zitatu zolembedwa ndi mawonekedwe ochiritsira amitundu isanu ndi iwiri omwe amagwiritsidwa ntchito mu phunziro lathu lapitalo akuwonetsedwa mkuyu. magalasi mu Chithunzi 1a.Sizithunzi zonse zakuthwa, kukula kwa chithunzi kumawonjezeka kuchokera ku C1 mpaka C7.Zithunzi makumi awiri mphambu zisanu ndi zitatu ndi 0.70 ± 0.19 mm kukula kwake.Choncho, n'zovuta kusunga kusamvana kwakukulu muzithunzi zonse.The coefficient of variable (CV) ya kukula kwa chithunzi 28 mu Chithunzi 5b inali 28%, pamene CV ya kukula kwa chithunzi 36 mu Chithunzi 5a inatsikira ku 9%.Zotsatira zomwe zili pamwambazi zikuwonetsa kuti mawonekedwe atsopano amitundu isanu ndi inayi sikuti amangowonjezera kuchuluka kwa mitundu yoyezera nthawi imodzi kuchokera pa zisanu ndi ziwiri mpaka zisanu ndi zinayi, komanso amakhala ndi chithunzi chapamwamba pamtundu uliwonse.
Kuyerekeza kwa mtundu wa chithunzi chogawanika chopangidwa ndi ma spectrometer ochiritsira komanso atsopano.(a) Magulu anayi azithunzi zolekanitsidwa zamitundu isanu ndi inayi (C1-C9) opangidwa ndi spectrometer yatsopano yamitundu isanu ndi inayi.(b) Maseti anayi a zithunzi zolekanitsidwa zamitundu isanu ndi iwiri (C1-C7) zopangidwa ndi mawonekedwe owoneka bwino amitundu isanu ndi iwiri.Ma Fluxes (C0) okhala ndi kutalika kwa mafunde kuchokera ku 400 mpaka 750 nm kuchokera kumalo anayi otulutsa amaphatikizidwa ndi zochitika pa spectrometer iliyonse, motsatana.
Mawonekedwe amtundu wa spectrometer wamitundu isanu ndi inayi adayesedwa moyesera ndipo zotsatira zowunikira zikuwonetsedwa mu Chithunzi 6. Onani kuti Chithunzi 6a chikuwonetsa zotsatira zomwezo monga Chithunzi 5a, mwachitsanzo pa mafunde a 4 C0 400-750 nm, zithunzi zonse za 36 zimadziwika. (Magulu 4 C1-C9).M'malo mwake, monga momwe tawonetsera mkuyu. magulu adazindikira C1, C2, C3, C4, C5, C6, C7, C8 kapena C9).Komabe, zithunzi zina zomwe zili pafupi ndi zithunzi zinayi zofananira zimadziwika mofooka kwambiri chifukwa C1-C9 transmission spectra yomwe ikuwonetsedwa mu Fig. 4b imadutsana pang'ono ndipo C0 iliyonse ili ndi gulu la 10 nm pamtunda wapadera monga momwe tafotokozera m'njira.Zotsatira izi zimagwirizana ndi mawonekedwe opatsirana a C1-C9 omwe akuwonetsedwa pazithunzi.4b ndi mavidiyo owonjezera 2 ndi 3. Mwa kuyankhula kwina, spectrometer yamitundu isanu ndi inayi imagwira ntchito monga momwe ikuyembekezeredwa kutengera zotsatira zomwe zasonyezedwa mkuyu.4b .Chifukwa chake, zimaganiziridwa kuti kugawa kwamphamvu kwa chithunzi C1-C9 ndiye mawonekedwe a C0 iliyonse.
Makhalidwe owoneka bwino a spectrometer yamitundu isanu ndi inayi.Sipekitiromita yatsopano yamitundu isanu ndi inayi imapanga ma seti anayi a zithunzi zolekanitsidwa zamitundu isanu ndi inayi (C1-C9) pomwe kuwala kwa chochitika (4 C0) kumakhala ndi kutalika kwa (a) 400-750 nm (monga momwe tawonetsera pa Chithunzi 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, motsatana.
Chojambula chojambula chamitundu isanu ndi inayi chinagwiritsidwa ntchito pa electrophoresis ya capillary inayi (kuti mudziwe zambiri, onani Zowonjezera Zowonjezera)31,34,35.Matrix a capillary anayi amakhala ndi ma capillaries anayi (m'mimba mwake 360 ​​μm ndi m'mimba mwake 50 μm) omwe amakhala pamtunda wa 1 mm pamalo owala a laser.Zitsanzo zomwe zili ndi zidutswa za DNA zolembedwa ndi utoto 8, zomwe ndi FL-6C (dye 1), JOE-6C (dye 2), dR6G (dye 3), TMR-6C (dye 4), CXR-6C (dye 5), TOM- 6C (dye 6), LIZ (dye 7), ndi WEN (dye 8) mu dongosolo lokwera la fluorescent wavelength, olekanitsidwa mumtundu uliwonse wa ma capillaries anayi (pambuyo pake amatchedwa Cap1, Cap2, Cap3, ndi Cap4).Laser-induced fluorescence yochokera ku Cap1-Cap4 idalumikizidwa ndi ma lens angapo ndipo nthawi imodzi idajambulidwa ndi spectrometer yamitundu isanu ndi inayi.Kukula kwamphamvu kwamitundu isanu ndi inayi (C1-C9) fluorescence pa electrophoresis, ndiko kuti, electrophoregram yamitundu isanu ndi inayi ya capillary iliyonse, ikuwonetsedwa mkuyu 7a.Electrophoregram yofanana yamitundu isanu ndi inayi imapezeka mu Cap1-Cap4.Monga momwe mivi ya Cap1 ikuwonetsera pa Chithunzi 7a, nsonga zisanu ndi zitatu pa electrophoregram iliyonse yamitundu isanu ndi inayi ikuwonetsa kutuluka kwa fulorosenti kuchokera ku Dye1-Dye8, motsatana.
Kuchulukitsidwa kofanana kwa utoto asanu ndi atatu pogwiritsa ntchito spectrometer yamitundu isanu ndi inayi ya capillary electrophoresis.(a) Electrophoregram yamitundu isanu ndi inayi (C1-C9) ya capillary iliyonse.Zisonga zisanu ndi zitatu zosonyezedwa ndi mivi Cap1 zikuwonetsa mpweya wa fluorescence wamitundu isanu ndi itatu (Dye1-Dye8).Mitundu ya mivi imagwirizana ndi mitundu (b) ndi (c).(b) Fluorescence spectra ya utoto eyiti (Dye1-Dye8) pa capillary.c Electropherograms ya utoto eyiti (Dye1-Dye8) pa capillary.Pamwamba pa zidutswa za DNA zotchedwa Dye7 zimasonyezedwa ndi mivi, ndipo kutalika kwake kwa Cap4 kumasonyezedwa.
Kugawidwa kwamphamvu kwa C1-C9 pamapiri asanu ndi atatu akuwonetsedwa muzithunzi.7b, pa.Chifukwa onse a C1-C9 ndi Dye1-Dye8 ali mu dongosolo la kutalika kwa mawonekedwe, magawo asanu ndi atatu mu Chithunzi 7b amasonyeza mawonekedwe a fluorescence a Dye1-Dye8 motsatizana kuchokera kumanzere kupita kumanja.M’kafukufukuyu, Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, ndi Dye8 amawoneka mwa magenta, violet, blue, cyan, green, yellow, orange, and red, motsatana.Zindikirani kuti mitundu ya mivi mumkuyu 7a imagwirizana ndi mitundu ya utoto mu Mkuyu 7b.Mphamvu ya C1-C9 fluorescence pa sipekitiramu iliyonse mu Chithunzi 7b idasinthidwa kuti chiŵerengero chawo chikhale chimodzi.Mawonekedwe asanu ndi atatu ofanana ndi fluorescence adapezedwa kuchokera ku Cap1-Cap4.Munthu amatha kuwona kuphatikizika kowoneka bwino kwa fluorescence pakati pa utoto wa 1-dye 8.
Monga momwe tawonetsera pa Chithunzi 7c, pamtundu uliwonse wa capillary, electrophoregram yamitundu isanu ndi inayi mu Chithunzi 7a inasinthidwa kukhala electropherogram yamitundu isanu ndi itatu ndi kusanthula kwamagulu ambiri pogwiritsa ntchito mawonekedwe asanu ndi atatu a fluorescence mu Chithunzi 7b (onani Zowonjezera Zowonjezera kuti mudziwe zambiri).Popeza kuphatikizika kwa mawonekedwe a fulorosenti mu Chithunzi 7a sikuwonetsedwa mu Chithunzi 7c, Dye1-Dye8 imatha kudziwika ndikuwerengedwa payekhapayekha nthawi iliyonse, ngakhale mitundu yosiyanasiyana ya Dye1-Dye8 fulorosi nthawi imodzi.Izi sizingachitike ndi kuzindikira kwamitundu isanu ndi iwiri31, koma zitha kutheka ndi kuzindikira kwamitundu isanu ndi inayi.Monga momwe mivi Cap1 mumkuyu 7c, fulorosenti emission singlets Dye3 (buluu), Dye8 (wofiira), Dye5 (wobiriwira), Dye4 (cyan), Dye2 (wofiirira), Dye1 (magenta), ndi Dye6 (Yellow). ) amaonedwa m’ndondomeko yoyembekezeredwa.Pakutulutsa fulorosenti ya utoto 7 (lalanje), kuwonjezera pa nsonga imodzi yowonetsedwa ndi muvi wa lalanje, nsonga zina zingapo zingapo zidawonedwa.Chotsatirachi ndi chifukwa chakuti zitsanzozo zinali ndi miyezo ya kukula, Dye7 yolembedwa zidutswa za DNA zokhala ndi utali wosiyana.Monga momwe tawonetsera pa Chithunzi 7c, kwa Cap4 kutalika kwa maziko awa ndi 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 ndi 220 kutalika kwa maziko.
Zinthu zazikulu za spectrometer yamitundu isanu ndi inayi, yopangidwa pogwiritsa ntchito matrix a magalasi amitundu iwiri ya dichroic, ndi yaying'ono komanso kapangidwe kosavuta.Popeza magalasi a decachromatic mkati mwa adaputala akuwonetsedwa mkuyu.3c yokwera mwachindunji pa bolodi la sensa ya chithunzi (onani Mkuyu. S1 ndi S2), spectrometer yamitundu isanu ndi inayi imakhala ndi miyeso yofanana ndi adaputala, mwachitsanzo 54 × 58 × 8.5 mm.(kukula).Kukula kwakung'ono kwambiriku ndi maoda awiri kapena atatu a kukula kwake kocheperako kuposa ma spectrometer wamba omwe amagwiritsa ntchito ma grating kapena ma prisms.Kuphatikiza apo, popeza spectrometer yamitundu isanu ndi inayi imapangidwa kotero kuti kuwala kumagunda pamwamba pa sensa ya chithunzi perpendicularly, danga litha kuperekedwa mosavuta kwa ma spectrometer amitundu isanu ndi inayi pamakina monga ma microscopes, ma flow cytometer, kapena analyzer.Capillary grating electrophoresis analyzer ya miniaturization yayikulu kwambiri ya dongosolo.Panthawi imodzimodziyo, kukula kwa magalasi khumi a dichroic ndi zosefera za bandpass zomwe zimagwiritsidwa ntchito mu spectrometer yamitundu isanu ndi inayi ndi 10 × 1.9 × 0.5 mm kapena 15 × 1.9 × 0.5 mm.Chifukwa chake, magalasi ang'onoang'ono opitilira 100 a dichroic ndi zosefera za bandpass, motsatana, zitha kudulidwa kuchokera pagalasi la dichroic ndi fyuluta ya 60 mm2 bandpass, motsatana.Choncho, magalasi angapo a decachromatic amatha kupangidwa pamtengo wotsika.
Chinthu china cha spectrometer yamitundu isanu ndi inayi ndi mawonekedwe ake abwino kwambiri.Makamaka, zimalola kupeza zithunzi zowoneka bwino zazithunzi, ndiko kuti, kupeza nthawi yomweyo zithunzi zokhala ndi chidziwitso chowoneka bwino.Pa chithunzi chilichonse, mawonekedwe osalekeza adapezedwa ndi kutalika kwa kutalika kwa 520 mpaka 700 nm komanso kusamvana kwa 20 nm.Mwa kuyankhula kwina, mphamvu zisanu ndi zinayi za kuwala zimadziwika pachithunzi chilichonse, mwachitsanzo, magulu asanu ndi anayi a 20 nm omwe amagawanitsa kutalika kwa mawonekedwe kuchokera ku 520 mpaka 700 nm.Posintha mawonekedwe a mawonekedwe a galasi la dichroic ndi fyuluta ya bandpass, kutalika kwa mawonekedwe a magulu asanu ndi anayi ndi m'lifupi mwa gulu lirilonse likhoza kusinthidwa.Kuzindikira mitundu isanu ndi inayi sikungagwiritsidwe ntchito poyeza miyeso ya fluorescence yokhala ndi zithunzi zowoneka bwino (monga momwe tafotokozera mu lipoti ili), komanso pazinthu zina zambiri zomwe zimagwiritsidwa ntchito pogwiritsa ntchito kujambula kowoneka bwino.Ngakhale kuti zithunzithunzi za hyperspectral zimatha kuzindikira mazana amitundu, zapezeka kuti ngakhale kuchepa kwakukulu kwa chiwerengero cha mitundu yodziwika bwino, zinthu zambiri zomwe zili m'mawonekedwe amatha kudziwika ndi kulondola kokwanira kwa mapulogalamu ambiri38,39,40.Chifukwa kusasunthika kwa malo, mawonekedwe owoneka bwino, komanso kusintha kwakanthawi kumakhala ndi kusinthana kwamalingaliro owoneka bwino, kuchepetsa kuchuluka kwa mitundu kumatha kusintha kusintha kwa malo ndikusintha kwakanthawi.Itha kugwiritsanso ntchito ma spectrometer osavuta ngati omwe apangidwa mu kafukufukuyu ndikuchepetsanso kuchuluka kwa kuwerengera.
Mu kafukufukuyu, mitundu isanu ndi itatu idawerengedwa nthawi imodzi ndikulekanitsa kowoneka bwino kwa mawonekedwe awo amtundu wa fluorescence kutengera kupezeka kwa mitundu isanu ndi inayi.Kufikira utoto zisanu ndi zinayi ukhoza kuwerengedwa nthawi imodzi, kukhalapo mu nthawi ndi malo.Ubwino wapadera wa spectrometer yamitundu isanu ndi inayi ndi kuwala kwake kowala komanso kabowo kakang'ono (1 × 7 mm).Gulu lagalasi la decane limakhala ndi kufalikira kwakukulu kwa 92% ya kuwala kuchokera pachibowo chilichonse mwa migawo isanu ndi inayi ya mafunde.Kuthekera kogwiritsa ntchito kuwala kochitika mumtundu wa wavelength kuchokera pa 520 mpaka 700 nm ndi pafupifupi 100%.Pamitundu yosiyanasiyana ya mafunde, palibe ma diffraction grating omwe angapereke mphamvu yogwiritsira ntchito.Ngakhale mphamvu ya diffraction ya grating ya diffraction iposa 90% pa utali wina wa wavelength, kusiyana pakati pa utali wa wavelength ndi utali winawake ukuwonjezeka, mphamvu ya diffraction pa utali wina imachepa41.The kabowo m'lifupi perpendicular malangizo ndege mu Mkuyu 2c akhoza anawonjezera kuchokera 7 mm kwa m'lifupi kachipangizo chithunzi, monga mu nkhani ya sensa chithunzi ntchito mu phunziroli, mwa kusintha pang'ono decamer array.
Ma spectrometer amitundu isanu ndi inayi angagwiritsidwe ntchito osati capillary electrophoresis, monga momwe tawonetsera mu phunziroli, komanso pazifukwa zina zosiyanasiyana.Mwachitsanzo, monga momwe tawonetsera m'chithunzichi, spectrometer yamitundu isanu ndi inayi ingagwiritsidwe ntchito pa microscope ya fluorescence.Ndege yachitsanzo ikuwonetsedwa pa sensa ya chithunzi cha spectrometer yamitundu isanu ndi inayi kudzera mu cholinga cha 10x.Mtunda wa kuwala pakati pa cholinga cha lens ndi sensa ya chithunzi ndi 200 mm, pamene mtunda wa kuwala pakati pa zochitika za spectrometer yamitundu isanu ndi inayi ndi sensa ya chithunzi ndi 12 mm yokha.Choncho, chithunzicho chinadulidwa pafupifupi kukula kwa kabowo (1 × 7 mm) mu ndege ya zochitika ndikugawidwa muzithunzi zisanu ndi zinayi.Ndiko kuti, chithunzi chowonekera cha chithunzi chamitundu isanu ndi inayi chikhoza kutengedwa pa 0.1 × 0.7 mm dera mu ndege yachitsanzo.Kuonjezera apo, n'zotheka kupeza chithunzi chamitundu isanu ndi inayi cha malo okulirapo pa ndege yachitsanzo poyang'ana chitsanzo chokhudzana ndi cholinga chotsatira chopingasa mu Mkuyu 2c.
Magalasi a decachromatic array, omwe ndi M1-M9 ndi BP, adapangidwa ndi Asahi Spectra Co., Ltd. pogwiritsa ntchito njira zodziwika bwino zamvula.Multilayer dielectric zipangizo zinagwiritsidwa ntchito payekha pa mbale khumi quartz 60 × 60 mm kukula ndi 0.5 mm wandiweyani, kukwaniritsa zofunika izi: M1: IA = 45 °, R ≥ 90% pa 520-590 nm, Tave ≥ 90% pa 610– 610 nm.700 nm, M2: IA = 45 °, R ≥ 90% pa 520-530 nm, Tave ≥ 90% pa 550-600 nm, M3: IA = 45 °, R ≥ 90% pa 540-550 nm, Tave 00 % pa 570-600 nm, M4: IA = 45 °, R ≥ 90% pa 560-570 nm, Tave ≥ 90% pa 590-600 nm, M5: IA = 45 °, R ≥ 98% pa 580-600 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 T0≥ 530 nm T0≥ 530 nm pa -690 nm ndi T ≤ 1% pa 725-750 nm, kumene IA, T, Tave, ndi R ndi ngodya ya zochitika, transmittance, transmittance avareji, ndi unpolarized kuwala kuwala.
Kuwala koyera (C0) kokhala ndi kutalika kwa mafunde a 400–750 nm opangidwa ndi gwero la kuwala kwa LED (AS 3000, AS ONE CORPORATION) kudawonjezedwa ndipo zidachitika molunjika pa DP ya magalasi osiyanasiyana osiyanasiyana.Kuwala koyera kwa ma LED kukuwonetsedwa mu Supplementary Figure S3.Ikani tanki ya acrylic (miyeso 150 × 150 × 30 mm) kutsogolo kwa galasi la decamera, moyang'anizana ndi PSU.Utsi wotuluka pamene ayezi wowuma umizidwa m'madzi kenako adatsanuliridwa mu thanki ya acrylic kuti muwone mitsinje yamitundu isanu ndi inayi ya C1-C9 yomwe imachokera ku magalasi angapo a decachromatic.
Kapenanso, kuwala koyera (C0) kumadutsa mu fyuluta musanalowe mu DP.Zosefera poyamba zinali zosefera osalowerera ndale zokhala ndi kachulukidwe kakang'ono ka 0.6.Kenako gwiritsani ntchito fyuluta yamoto (FW212C, FW212C, Thorlabs).Pomaliza, yatsaninso fyuluta ya ND.Ma bandwidths a zosefera zisanu ndi zinayi za bandpass zimagwirizana ndi C9, C8, C7, C6, C5, C4, C3, C2 ndi C1, motsatana.Selo la quartz lokhala ndi miyeso yamkati ya 40 (kutalika kwa kuwala) x 42.5 (kutalika) x 10 mm (m'lifupi) inayikidwa patsogolo pa magalasi a decochromatic, moyang'anizana ndi BP.Utsiwo umadyetsedwa kudzera mu chubu kulowa mu cell ya quartz kuti utsi ukhale wochuluka mu selo la quartz kuti muwone mitsinje yamitundu isanu ndi inayi ya C1-C9 yomwe imachokera ku galasi la decachromatic.
Kanema wa mtsinje wamitundu isanu ndi inayi wogawanika kuchokera ku magalasi angapo a decanic adajambulidwa mumayendedwe anthawi yayitali pa iPhone XS.Jambulani zithunzi za zochitika pa 1 fps ndikuphatikiza zithunzizo kuti mupange kanema pa 30 fps (pa kanema wosasankha 1) kapena 24 fps (pamavidiyo osasankha 2 ndi 3).
Ikani mbale yachitsulo yosapanga dzimbiri 50 µm (yokhala ndi mabowo anayi a 50 µm m'mimba mwake pa 1 mm intervals) pa mbale ya diffusion.Kuwala kokhala ndi kutalika kwa 400-750 nm kumawunikiridwa pa mbale ya diffuser, yomwe imapezedwa podutsa kuwala kuchokera ku nyali ya halogen kudzera pa fyuluta yaying'ono yokhala ndi kutalika kwa 700 nm.Kuwala kowala kukuwonetsedwa mu Supplementary Figure S4.Kapenanso, kuwala kumadutsanso imodzi mwazosefera za 10 nm bandpass zomwe zili pa 530, 550, 570, 590, 610, 630, 650, 670 ndi 690 nm ndikugunda mbale ya diffuser.Zotsatira zake, ma radiation anayi okhala ndi mainchesi a φ50 μm ndi mafunde osiyanasiyana adapangidwa pa mbale yachitsulo chosapanga dzimbiri moyang'anizana ndi mbale ya diffuser.
Gulu la ma capillary anayi okhala ndi ma lens anayi amayikidwa pa spectrometer yamitundu isanu ndi inayi monga momwe zikusonyezedwera mu Zithunzi 1 ndi 2. C1 ndi C2.Ma capillaries anayi ndi ma lens anayi anali ofanana ndi maphunziro apitalo31,34.Mtengo wa laser wokhala ndi kutalika kwa 505 nm ndi mphamvu ya 15 mW umawunikiridwa nthawi imodzi komanso mofanana kuchokera kumbali kupita kumalo otulutsa ma capillaries anayi.Fluorescence yotulutsidwa ndi malo aliwonse otulutsa imalumikizidwa ndi lens yofananira ndikugawanika kukhala mitsinje isanu ndi inayi ndi magalasi angapo a decachromatic.Mitsinje ya 36 yotsatiridwayo idalowetsedwa mwachindunji mu sensa ya chithunzi cha CMOS (C11440-52U, Hamamatsu Photonics K · K.), ndipo zithunzi zawo zidajambulidwa nthawi imodzi.
ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Applied Biosystems), 4 µl GeneScan™ 600 LIZ™ utoto unasakanizidwa pa capillary iliyonse posakaniza 1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), 1 µl mix size standard.v2.0 (Thermo Fisher Scientific) ndi 14 µl madzi.PowerPlex® 6C Matrix Standard ili ndi zidutswa zisanu ndi chimodzi za DNA zolembedwa ndi utoto 6: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, ndi WEN, potengera kutalika kwa kutalika kwake.Kutalika kwa maziko a zidutswa za DNA izi sizikuwululidwa, koma kutalika kwa maziko a zidutswa za DNA zolembedwa ndi WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C ndi TOM-6C zimadziwika.Zosakaniza zomwe zili mu ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit zili ndi chidutswa cha DNA cholembedwa utoto wa dR6G.Kutalika kwa maziko a zidutswa za DNA sikudziwikanso.GeneScan™ 600 LIZ™ Dye Size Standard v2.0 imaphatikizapo zidutswa 36 za DNA zolembedwa ndi LIZ.Kutalika kwa zigawo za DNA izi ndi 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 40, 30, 30, 30 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 ndi 600 maziko.Zitsanzozo zidasinthidwa pa 94 ​​° C kwa mphindi zitatu, kenako zitakhazikika pa ayezi kwa mphindi zisanu.Zitsanzo zidalowetsedwa mu capillary iliyonse pa 26 V/cm kwa 9 s ndikusiyanitsidwa mu capillary iliyonse yodzazidwa ndi POP-7™ polima solution (Thermo Fisher Scientific) yokhala ndi kutalika kwa 36 cm ndi voliyumu ya 181 V/cm ndi mbali ya 60 °.KUCHOKERA.
Deta yonse yomwe yapezedwa kapena kufufuzidwa panthawi ya phunziroli ikuphatikizidwa m'nkhani yofalitsidwayi ndi zina zowonjezera.Zina zomwe zikugwirizana ndi kafukufukuyu zikupezeka kuchokera kwa olemba omwe afunsidwa ngati afunsidwa.
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