Stainless hlau 321 8 * 1.2 coiled raj rau tshav kub exchanger

Capillary Tubes

Ua tsaug rau koj tuaj xyuas Nature.com.Koj siv lub browser version nrog kev txhawb nqa CSS tsawg.Rau qhov kev paub zoo tshaj plaws, peb xav kom koj siv qhov browser tshiab (lossis lov tes taw Compatibility Hom hauv Internet Explorer).Tsis tas li ntawd, txhawm rau xyuas kom muaj kev txhawb nqa tsis tu ncua, peb qhia lub vev xaib tsis muaj qauv thiab JavaScript.
Qhia ib lub carousel ntawm peb slides ib zaug.Siv cov nyees khawm dhau los thiab Tom ntej kom txav mus los ntawm peb qhov swb ib zaug, lossis siv cov khawm slider thaum kawg kom txav mus los ntawm peb qhov swb ib zaug.
Ib qho ultra-compact (54 × 58 × 8.5 mm) thiab dav-aperture (1 × 7 mm) cuaj xim spectrometer tau tsim, "sib cais hauv ob" los ntawm ib qho ntawm kaum daim iav dichroic, uas tau siv rau cov duab spectral instantaneous.Qhov teeb meem lub teeb flux nrog ib ntu me dua qhov aperture loj yog muab faib ua ib qho txuas txuas ntxiv 20 nm dav thiab cuaj xim flux nrog nruab nrab wavelengths ntawm 530, 550, 570, 590, 610, 630, 650, 670 thiab 690 nm.Cov duab ntawm cuaj xim kwj yog ib txhij ntsuas tau zoo los ntawm cov duab sensor.Tsis zoo li cov qauv dichroic daim iav arrays, cov tsim dichroic daim iav array muaj ib tug tshwj xeeb ob-thooj configuration, uas tsis tsuas yog nce tus naj npawb ntawm cov xim uas yuav ntsuas ib txhij, tab sis kuj txhim kho cov duab daws teeb meem rau txhua xim kwj.Tus tsim cuaj-xim spectrometer yog siv rau plaub-capillary electrophoresis.Simultaneous quantitative tsom xam ntawm yim dyes migrating ib txhij nyob rau hauv txhua capillary siv cuaj-xim laser-induced fluorescence.Txij li thaum lub cuaj-xim spectrometer tsis yog tsuas yog ultra-me me thiab pheej yig, tab sis kuj muaj siab luminous flux thiab txaus spectral daws teeb meem rau feem ntau spectral imaging daim ntaub ntawv, nws yuav siv tau nyob rau hauv ntau yam teb.
Hyperspectral thiab multispectral imaging tau dhau los ua ib feem tseem ceeb ntawm astronomy2, chaw taws teeb rau kev soj ntsuam lub ntiaj teb 3,4, kev tswj cov khoom noj thiab dej zoo5,6, kev txuag kos duab thiab archaeology7, forensics8, phais9, biomedical analysis thiab diagnostics10,11 thiab lwm yam. Field 1 Ib qho indispensable technology , 12, 13.Cov txheej txheem rau kev ntsuas lub spectrum ntawm lub teeb emissions los ntawm txhua lub ntsiab lus ntawm emission nyob rau hauv qhov kev pom yog muab faib ua (1) point scanning ("broom") 14,15, (2) linear scanning ("panicle") 16,17,18 , (3) ntev scans waves19,20,21 thiab (4) images22,23,24,25.Nyob rau hauv cov ntaub ntawv ntawm tag nrho cov txheej txheem no, spatial daws teeb meem, spectral daws teeb meem thiab lub cev kev daws teeb meem muaj kev sib pauv-tawm kev sib raug zoo9,10,12,26.Tsis tas li ntawd, lub teeb tso zis muaj qhov cuam tshuam tseem ceeb ntawm qhov rhiab heev, piv txwv li lub teeb liab-rau-lub suab sib piv hauv spectral imaging26.Lub luminous flux, uas yog, qhov ua tau zoo ntawm kev siv lub teeb, yog ncaj qha proportional rau qhov sib piv ntawm qhov ntsuas qhov tseeb ntawm lub teeb ntawm txhua qhov luminous point ntawm ib chav tsev lub sij hawm mus rau tag nrho cov teeb ntawm lub ntsuas wavelength.Qeb (4) yog ib txoj hauv kev tsim nyog thaum qhov kev siv los yog spectrum ntawm lub teeb emission los ntawm txhua qhov emitting point hloov nrog lub sij hawm los yog thaum lub sij hawm txoj hauj lwm ntawm txhua emitting point hloov nrog lub sij hawm vim hais tias lub spectrum ntawm lub teeb emitted los ntawm tag nrho cov emitting ntsiab lus yog ntsuas ib txhij.24.
Feem ntau ntawm cov txheej txheem saum toj no yog ua ke nrog loj, nyuaj thiab / lossis kim spectrometers siv 18 gratings lossis 14, 16, 22, 23 prisms rau cov chav kawm (1), (2) thiab (4) lossis 20, 21 lim disks, kua lim .Crystalline tunable lim (LCTF)25 lossis acousto-optic tunable lim (AOTF)19 ntawm qeb (3).Hauv qhov sib piv, qeb (4) ntau daim iav spectrometers me me thiab pheej yig vim lawv qhov yooj yim configuration27,28,29,30.Tsis tas li ntawd, lawv muaj lub siab luminous flux vim hais tias lub teeb sib koom los ntawm txhua daim iav dichroic (uas yog, lub teeb xa tawm thiab cuam tshuam ntawm qhov teeb meem teeb pom kev zoo ntawm txhua daim iav dichroic) yog siv tas li thiab tsis tu ncua.Txawm li cas los xij, tus naj npawb ntawm wavelength bands (xws li xim) uas yuav tsum tau ntsuas ib txhij yog txwv rau plaub.
Spectral imaging raws li kev kuaj pom fluorescence feem ntau yog siv rau kev tsom xam multiplex hauv biomedical detection thiab diagnostics 10, 13 .Hauv multiplexing, txij li ntau qhov kev txheeb xyuas (piv txwv li, DNA tshwj xeeb lossis cov proteins) tau sau nrog cov fluorescent dyes sib txawv, txhua qhov kev ntsuas tam sim no ntawm txhua qhov emission point nyob rau hauv qhov kev pom yog kom muaj nuj nqis siv ntau qhov kev tshuaj ntsuam.32 rhuav tshem cov fluorescence spectrum uas pom los ntawm txhua qhov emission point.Thaum lub sijhawm txheej txheem no, cov xim sib txawv, txhua qhov sib txawv ntawm cov fluorescence tuaj yeem ua rau colocalize, uas yog, sib koom ua ke hauv qhov chaw thiab lub sijhawm.Tam sim no, qhov siab tshaj plaws ntawm cov dyes uas tuaj yeem txaus siab los ntawm ib lub laser beam yog yim33.Qhov kev txwv sab saud no tsis yog txiav txim siab los ntawm qhov kev daws teeb meem spectral (piv txwv li, pes tsawg xim), tab sis los ntawm qhov dav ntawm fluorescence spectrum (≥50 nm) thiab tus nqi dye Stokes hloov (≤200 nm) ntawm FRET (siv FRET) 10 .Txawm li cas los xij, tus naj npawb ntawm cov xim yuav tsum ntau dua lossis sib npaug ntawm cov dyes kom tshem tawm cov spectral sib tshooj ntawm cov dyes31,32.Yog li ntawd, nws yog ib qho tsim nyog yuav tsum tau nce cov xim ib txhij ntsuas rau yim lossis ntau dua.
Tsis ntev los no, ib qho ultra-compact heptachroic spectrometer (siv ib qho array ntawm heptychroic iav thiab ib qho duab sensor los ntsuas plaub fluorescent fluxes) tau tsim.Lub spectrometer yog ob mus rau peb qhov kev txiav txim ntawm qhov loj me dua li cov qauv spectrometers siv gratings lossis prisms34,35.Txawm li cas los xij, nws nyuaj rau tso ntau dua xya lub iav dichroic hauv ib lub spectrometer thiab ib txhij ntsuas ntau dua xya xim36,37.Nrog rau qhov nce ntawm cov iav dichroic, qhov siab tshaj plaws qhov sib txawv ntawm qhov ntev ntawm txoj kev kho qhov muag ntawm dichroic lub teeb fluxes nce, thiab nws nyuaj rau tso saib tag nrho lub teeb fluxes ntawm ib lub dav hlau sensory.Qhov ntev tshaj plaws optical txoj kev ntev ntawm lub teeb flux kuj nce, yog li qhov dav ntawm lub spectrometer aperture (piv txwv li qhov siab tshaj plaws ntawm lub teeb tsom los ntawm spectrometer) txo qis.
Hauv kev teb rau cov teeb meem saum toj no, lub ultra-compact cuaj-xim spectrometer nrog ob txheej "dichroic" decachromatic daim iav array thiab ib qho duab sensor rau instantaneous spectral imaging [qeb (4)] tau tsim.Piv nrog rau yav dhau los spectrometers, tus tsim spectrometer muaj qhov sib txawv me me nyob rau hauv qhov siab tshaj plaws kho qhov muag txoj kev ntev thiab ib tug me me qhov siab tshaj plaws optical txoj kev ntev.Nws tau raug thov rau plaub-capillary electrophoris los ntes cov laser-indicescence thiab kom ntsuas lub fluorescence thiab kom paub tseeb kev tsiv teb chaws ntawm yim tus tuag hauv txhua lub capillary.Txij li thaum tsim spectrometer tsis yog tsuas yog ultra-me me thiab pheej yig, tab sis kuj muaj lub siab luminous flux thiab txaus spectral daws teeb meem rau feem ntau spectral imaging daim ntaub ntawv, nws yuav siv tau nyob rau hauv ntau yam teb.
Cov tsoos cuaj-xim spectrometer yog qhia hauv daim duab.1 ib a.Nws tsim raws li cov yav dhau los ultra-me me xya-xim spectrometer 31. Nws muaj cuaj dichroic tsom iav teem kab rov tav ntawm lub kaum sab xis ntawm 45 ° mus rau sab xis, thiab cov duab sensor (S) nyob saum cuaj dichroic iav.Lub teeb nkag los hauv qab (C0) muab faib los ntawm ib qho array ntawm cuaj dichroic tsom iav rau cuaj lub teeb ntws mus (C1, C2, C3, C4, C5, C6, C7, C8 thiab C9).Tag nrho cuaj xim kwj yog pub ncaj qha rau cov duab sensor thiab raug kuaj pom ib txhij.Hauv txoj kev tshawb no, C1, C2, C3, C4, C5, C6, C7, C8, thiab C9 yog nyob rau hauv kev txiav txim ntawm wavelength thiab yog sawv cev los ntawm magenta, violet, xiav, cyan, ntsuab, daj, txiv kab ntxwv, liab-txiv kab ntxwv, thiab liab, feem.Txawm hais tias cov xim no siv rau hauv daim ntawv no, raws li pom hauv daim duab 3, vim tias lawv txawv ntawm cov xim tiag tiag pom los ntawm tib neeg lub qhov muag.
Schematic daim duab ntawm cov pa thiab tshiab cuaj-xim spectrometers.(a) Cov pa cua cuaj-xim spectrometer nrog ib qho ntawm cuaj daim iav dichroic.(b) Tshiab cuaj-xim spectrometer nrog ob txheej dichroic daim iav array.Qhov xwm txheej lub teeb flux C0 tau muab faib ua cuaj xim lub teeb fluxes C1-C9 thiab kuaj pom los ntawm cov duab sensor S.
Tus tsim tshiab cuaj-xim spectrometer muaj ob txheej dichroic daim iav grating thiab ib qho duab sensor, raws li qhia hauv daim duab 1b.Hauv qib qis, tsib daim iav dichroic yog tilted 45 ° rau sab xis, ua raws rau sab xis los ntawm qhov nruab nrab ntawm cov array ntawm decamers.Nyob rau theem sab saum toj, tsib daim iav dichroic ntxiv yog tilted 45 ° rau sab laug thiab nyob ntawm qhov chaw mus rau sab laug.Daim iav sab laug tshaj plaws dichroic ntawm txheej qis thiab sab xis ntawm daim iav dichroic ntawm txheej sab sauv sib tshooj.Qhov teeb meem lub teeb flux (C0) tau muab faib los ntawm hauv qab no rau hauv plaub lub chromatic fluxes (C1-C4) los ntawm tsib lub iav dichroic ntawm sab xis thiab tsib lub chromatic fluxes (C5-C4) los ntawm tsib lub iav dichroic ntawm sab laug C9).Zoo li cov qauv cuaj-xim spectrometers, tag nrho cuaj xim ntws ncaj qha mus rau hauv cov duab sensor (S) thiab kuaj pom ib txhij.Sib piv cov duab 1a thiab 1b, ib tus tuaj yeem pom tias nyob rau hauv cov ntaub ntawv ntawm cuaj-xim spectrometer tshiab, ob qho tib si qhov sib txawv siab tshaj plaws thiab qhov ntev tshaj plaws kho qhov muag qhov ntev ntawm cuaj xim fluxes yog halved.
Cov ncauj lus kom ntxaws kev tsim kho ntawm ib qho ultra-me me ob-txheej dichroic daim iav array 29 mm (dav) × 31 mm (qhov tob) × 6 mm (qhov siab) yog qhia nyob rau hauv daim duab 2. Lub decimal dichroic daim iav array muaj tsib dichroic iav ntawm sab xis. (M1-M5) thiab tsib daim iav dichroic nyob rau sab laug (M6-M9 thiab lwm tus M5), txhua daim iav dichroic yog tsau nyob rau sab saum toj aluminium bracket.Tag nrho cov iav dichroic yog staggered kom them nyiaj rau qhov sib npaug ntawm qhov sib npaug vim qhov cuam tshuam ntawm qhov ntws los ntawm cov iav.Hauv qab M1, lub lim band-pass (BP) yog tsau.M1 thiab BP qhov ntev yog 10mm (ntev sab) x 1.9mm (sab luv) x 0.5mm (thickness).Qhov ntev ntawm qhov seem dichroic iav yog 15 hli × 1.9 hli × 0.5 hli.Lub matrix suab ntawm M1 thiab M2 yog 1.7 hli, thaum lub suab matrix ntawm lwm cov iav dichroic yog 1.6 hli.Ntawm daim duab.2c sib txuas cov teeb meem teeb pom kev zoo C0 thiab cuaj xim lub teeb fluxes C1-C9, sib cais los ntawm de-chamber matrix ntawm iav.
Kev tsim kho ntawm ob txheej dichroic daim iav matrix.(a) Kev pom kev pom thiab (b) ib qho kev saib hla ntawm ob txheej dichroic daim iav array (qhov loj me 29 hli x 31 hli x 6 hli).Nws muaj tsib lub iav dichroic (M1-M5) nyob rau hauv qis txheej, tsib lub iav dichroic (M6-M9 thiab lwm tus M5) nyob rau hauv lub sab sauv txheej, thiab ib tug bandpass lim (BP) nyob rau hauv qab M1.(c) Hla-sectional saib nyob rau hauv ntsug kev taw qhia, nrog C0 thiab C1-C9 sib tshooj.
Qhov dav ntawm lub aperture nyob rau hauv kab rov tav kev taw qhia, qhia los ntawm qhov dav C0 nyob rau hauv daim duab 2, c, yog 1 hli, thiab nyob rau hauv cov kev taw qhia perpendicular mus rau lub dav hlau daim duab 2, c, muab los ntawm tus tsim ntawm aluminium bracket, - 7 mm.Ntawd yog, cuaj-xim spectrometer tshiab muaj qhov loj aperture ntawm 1 hli × 7 hli.Txoj kev kho qhov muag ntawm C4 yog qhov ntev tshaj plaws ntawm C1-C9, thiab txoj kev kho qhov muag ntawm C4 hauv daim iav dichroic array, vim yog qhov loj me me (29 hli × 31 hli × 6 hli), yog 12 hli.Nyob rau tib lub sijhawm, txoj kev kho qhov muag ntev ntawm C5 yog qhov luv tshaj ntawm C1-C9, thiab txoj kev kho qhov muag ntev ntawm C5 yog 5.7 hli.Yog li ntawd, qhov siab tshaj plaws qhov sib txawv ntawm txoj kev kho qhov muag ntev yog 6.3 hli.Txoj kev kho qhov muag saum toj no raug kho rau qhov ntev ntawm txoj kev kho qhov muag rau kev sib kis ntawm qhov muag ntawm M1-M9 thiab BP (los ntawm quartz).
Lub spectral zog ntawm М1-М9 thiab VR yog xam kom cov fluxes С1, С2, С3, С4, С5, С6, С7, С8 thiab С9 nyob rau hauv lub wavelength ntau 520–540, 540–560, 560–580, 580 -600, 600–620, 620–640, 640–660, 660–680, thiab 680–700 nm, feem.
Daim duab ntawm cov khoom tsim matrix ntawm decachromatic iav yog qhia nyob rau hauv daim duab 3a.M1-M9 thiab BP yog glued mus rau 45 ° txoj kab nqes thiab kab rov tav dav hlau ntawm aluminium txhawb, feem, thaum M1 thiab BP muab zais rau sab nraum qab ntawm daim duab.
Kev tsim khoom ntawm ib qho array ntawm decan iav thiab nws cov qauv qhia.(a) Ib qho array ntawm fabricated decachromatic iav.(b) Ib daim duab 1 hli × 7 hli cuaj xim sib cais rau hauv ib daim ntawv tso rau pem hauv ntej ntawm ib qho array ntawm decachromatic iav thiab backlit nrog lub teeb dawb.(c) Ib qho ntawm cov iav decochromatic illuminated nrog lub teeb dawb ntawm qab.(d) Cuaj-xim splitting kwj emanating los ntawm decane daim iav array, pom los ntawm muab ib tug pa luam yeeb acrylic canister nyob rau hauv pem hauv ntej ntawm lub decane daim iav array ntawm c thiab darkening chav.
Kev ntsuas kis tau tus mob ntawm M1-M9 C0 ntawm lub kaum sab xis ntawm qhov xwm txheej ntawm 45 ° thiab ntsuas kev sib kis ntawm BP C0 ntawm lub kaum sab xis ntawm qhov xwm txheej ntawm 0 ° tau qhia hauv daim duab.4 ib a.Qhov kis tau tus mob ntawm C1-C9 txheeb ze rau C0 tau pom hauv daim duab.4b ib.Cov spectra no tau suav los ntawm spectra hauv Fig.4a raws li txoj kev kho qhov muag C1-C9 hauv daim duab 4a.1 ib 2c.Piv txwv li, 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)], where TS(X) and [ 1 − TS(X)] is the transmission and reflection spectra of X, ntsig.Raws li pom hauv daim duab 4b, bandwidths (bandwidth ≥50%) ntawm C1, C2, C3, C4, C5, C6, C7, C8 thiab C9 yog 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 thiab 682-699 nm.Cov txiaj ntsig no zoo ib yam nrog cov kab tsim kho.Tsis tas li ntawd, qhov kev siv ntawm C0 lub teeb yog siab, uas yog, qhov nruab nrab siab tshaj plaws C1-C9 lub teeb xa tawm yog 92%.
Kev kis tus kab mob ntawm daim iav dichroic thiab cais cuaj-xim flux.(a) Kev ntsuas kev sib kis ntawm M1-M9 ntawm 45 ° tshwm sim thiab BP ntawm 0 ° tshwm sim.(b) Transmission spectra of C1–C9 txheeb ze rau C0 xam los ntawm (a).
Ntawm daim duab.3c, lub array ntawm dichroic tsom iav yog nyob rau hauv vertically, yog li ntawd nws sab xis nyob rau hauv daim duab 3a yog sab saum toj thiab cov nqaj dawb ntawm lub collimated LED (C0) yog backlit.Lub array ntawm decachromatic iav pom nyob rau hauv daim duab 3a yog mounted nyob rau hauv ib tug 54 mm (qhov siab) × 58 mm (qhov tob) × 8.5 mm (thickness) adapter.Ntawm daim duab.3d, ntxiv rau lub xeev qhia hauv daim duab.3c, lub tank acrylic uas muaj pa luam yeeb tau muab tso rau pem hauv ntej ntawm ib qho array ntawm decochromatic tsom iav, nrog rau cov teeb hauv chav kaw.Yog li ntawd, cuaj dichroic ntws yog pom nyob rau hauv lub tank, emanating los ntawm ib tug array ntawm decachromatic iav.Txhua qhov kev sib cais muaj qhov sib npaug ntawm qhov sib npaug nrog qhov ntev ntawm 1 × 7 mm, uas sib haum rau qhov loj me ntawm cov cuaj xim spectrometer tshiab.Nyob rau hauv daim duab 3b, ib daim ntawv yog muab tso rau pem hauv ntej ntawm lub array ntawm dichroic tsom iav nyob rau hauv daim duab 3c, thiab ib tug 1 x 7 mm duab ntawm cuaj dichroic kwj projected rau daim ntawv yog pom los ntawm cov kev taw qhia ntawm daim ntawv txav.ntws.Lub cuaj xim sib cais ntws hauv daim duab.3b thiab d yog C4, C3, C2, C1, C5, C6, C7, C8 thiab C9 los ntawm sab saum toj mus rau hauv qab, uas tuaj yeem pom hauv cov duab 1 thiab 2. 1b thiab 2c.Lawv raug pom nyob rau hauv cov xim sib xws rau lawv cov wavelengths.Vim lub teeb dawb dawb siv ntawm LED (saib Ntxiv Fig. S3) thiab rhiab heev ntawm lub koob yees duab xim siv los ntes C9 (682–699 nm) hauv daim duab.Ib yam li ntawd, C9 tau pom faintly pom ntawm qhov muag liab qab.Lub caij no, C2 (kwj thib ob los ntawm sab saum toj) zoo nkaus li ntsuab hauv daim duab 3, tab sis zoo li daj ntau rau qhov muag liab qab.
Txoj kev hloov ntawm daim duab 3c mus rau d yog qhia hauv Cov Video Ntxiv 1. Tam sim ntawd tom qab lub teeb dawb ntawm LED hla dhau ntawm daim iav decachromatic, nws faib ib txhij mus rau cuaj xim.Thaum kawg, cov pa luam yeeb hauv lub vat maj mam dissiped los ntawm sab saum toj mus rau hauv qab, kom cuaj xim hmoov kuj ploj ntawm sab saum toj mus rau hauv qab.Nyob rau hauv sib piv, nyob rau hauv Ntxiv Video 2, thaum lub wavelength ntawm lub teeb flux tshwm sim ntawm lub array ntawm decachromatic tsom iav tau hloov los ntawm ntev mus rau luv nyob rau hauv qhov kev txiav txim ntawm 690, 671, 650, 632, 610, 589, 568, 550 thiab 532 nm ., Tsuas yog cov kab sib cais ntawm cuaj qhov sib cais hauv qhov kev txiav txim ntawm C9, C8, C7, C6, C5, C4, C3, C2, thiab C1.Acrylic reservoir yog hloov los ntawm lub pas dej ua ke quartz, thiab cov flakes ntawm txhua qhov shunted ntws yuav pom meej meej los ntawm txoj kev nqes hav.Tsis tas li ntawd, sub-video 3 yog edited xws li cov wavelength hloov ib feem ntawm cov sub-video 2 yog replayed.Qhov no yog qhov qhia tau zoo tshaj plaws ntawm cov yam ntxwv ntawm decochromatic array ntawm iav.
Cov txiaj ntsig saum toj no qhia tau hais tias cov khoom tsim decachromatic daim iav array lossis cuaj-xim spectrometer tshiab ua haujlwm raws li qhov xav tau.Lub cuaj-xim spectrometer tshiab yog tsim los ntawm mounting ib array ntawm decachromatic tsom iav nrog adapters ncaj qha mus rau cov duab sensor board.
Luminous flux nrog lub wavelength ntau ntawm 400 mus rau 750 nm, emitted los ntawm plaub lub ntsiab lus hluav taws xob φ50 μm, nyob ntawm 1 hli intervals nyob rau hauv cov kev taw qhia perpendicular mus rau lub dav hlau daim duab 2c, raws li kev tshawb fawb 31, 34. Cov plaub-lens array muaj plaub lo ntsiab muag φ1 hli nrog qhov ntev ntawm 1.4 hli thiab lub suab ntawm 1 hli.Plaub qhov sib tsoo (plaub C0) yog qhov xwm txheej ntawm DP ntawm cuaj-xim spectrometer tshiab, sib nrug ntawm 1 hli ntu.Ib qho array ntawm dichroic iav faib txhua kwj (C0) mus rau cuaj xim kwj (C1-C9).Qhov tshwm sim 36 kwj (plaub txheej ntawm C1-C9) yog tom qab ntawd txhaj ncaj qha rau hauv CMOS (S) duab sensor ncaj qha txuas mus rau ib qho ntawm cov iav dichroic.Yog li ntawd, raws li pom nyob rau hauv daim duab 5a, vim qhov me me qhov siab tshaj plaws kho qhov muag txoj kev sib txawv thiab txoj kev luv tshaj plaws kho qhov muag, cov duab ntawm tag nrho 36 kwj dej tau kuaj pom ib txhij thiab meej nrog qhov loj me.Raws li tus downstream spectra (saib daim duab ntxiv S4), cov duab siv ntawm plaub pawg C1, C2 thiab C3 kuj tsawg.Peb caug-rau cov duab yog 0.57 ± 0.05 hli hauv qhov loj me (txhais tau tias ± SD).Yog li, cov duab magnification nruab nrab yog 11.4.Qhov sib nrug ntsug ntawm cov duab nruab nrab yog 1 hli (tib qhov sib nrug raws li lub lens array) thiab kab rov tav qhov nruab nrab yog 1.6 hli (tib qhov sib nrug raws li daim iav dichroic array).Vim tias daim duab loj me dua li qhov kev ncua deb ntawm cov duab, txhua daim duab tuaj yeem ntsuas nws tus kheej (nrog rau crosstalk tsawg).Lub caij no, cov duab ntawm nees nkaum yim kwj sau tseg los ntawm cov pa xya-xim spectrometer siv hauv peb txoj kev tshawb fawb yav dhau los tau qhia hauv daim duab 5 B. Cov array ntawm xya lub iav dichroic tau tsim los ntawm kev tshem tawm ob daim iav dichroic ntawm lub array ntawm cuaj dichroic. iav hauv daim duab 1a.Tsis yog tag nrho cov duab yog ntse, cov duab loj nce ntawm C1 txog C7.Nees nkaum yim cov duab yog 0.70 ± 0.19 hli.Yog li, nws yog ib qho nyuaj rau tswj kom muaj kev daws teeb meem siab hauv txhua cov duab.Tus coefficient ntawm kev hloov pauv (CV) rau daim duab loj 28 hauv daim duab 5b yog 28%, thaum CV rau daim duab loj 36 hauv daim duab 5a poob rau 9%.Cov txiaj ntsig saum toj no qhia tau tias tus tshiab cuaj xim spectrometer tsis tsuas yog nce cov xim ib txhij ntsuas ntawm xya mus rau cuaj, tab sis kuj muaj cov duab siab daws teeb meem rau txhua xim.
Kev sib piv ntawm qhov zoo ntawm cov duab sib cais tsim los ntawm cov pa thiab cov spectrometers tshiab.(a) Plaub pawg ntawm cuaj xim sib cais cov duab (C1-C9) tsim los ntawm cuaj-xim spectrometer tshiab.(b) Plaub txheej ntawm xya-xim sib cais dluab (C1-C7) tsim nrog ib tug pa xya-xim spectrometer.Fluxes (C0) nrog wavelengths los ntawm 400 mus rau 750 nm los ntawm plaub lub ntsiab lus emission yog collimated thiab teeb meem ntawm txhua spectrometer, raws li.
Cov yam ntxwv spectral ntawm cuaj-xim spectrometer raug soj ntsuam sim thiab cov txiaj ntsig kev ntsuas tau pom hauv daim duab 6. Nco ntsoov tias daim duab 6a qhia tau zoo ib yam li daim duab 5a, piv txwv li ntawm wavelengths ntawm 4 C0 400-750 nm, tag nrho 36 cov duab raug kuaj pom. (4 pawg C1–C9).Ntawm qhov tsis sib xws, raws li qhia hauv daim duab 6b-j, thaum txhua C0 muaj qhov tshwj xeeb wavelength ntawm 530, 550, 570, 590, 610, 630, 650, 670, lossis 690 nm, yuav luag tsuas yog plaub daim duab sib xws (plaub pab pawg kuaj pom C1, C2, C3, C4, C5, C6, C7, C8 lossis C9).Txawm li cas los xij, qee cov duab uas nyob ib sab ntawm plaub cov duab sib thooj yog qhov tsis muaj zog heev vim tias C1-C9 kis tau tus mob spectra qhia hauv daim duab 4b sib tshooj me ntsis thiab txhua C0 muaj 10 nm band ntawm ib lub wavelength tshwj xeeb raws li tau piav qhia hauv txoj kev.Cov txiaj ntsig no zoo ib yam nrog C1-C9 kis tau tus mob spectra qhia hauv daim duab.4b thiab cov yeeb yaj kiab ntxiv 2 thiab 3. Hauv lwm lo lus, cuaj xim spectrometer ua haujlwm raws li qhov xav tau raws li cov txiaj ntsig tau pom hauv daim duab.4b ib.Yog li ntawd, nws tau txiav txim siab tias cov duab siv faib C1-C9 yog qhov spectrum ntawm txhua C0.
Spectral yam ntxwv ntawm cuaj-xim spectrometer.Lub cuaj-xim spectrometer tshiab tsim plaub pawg ntawm cuaj xim sib cais cov duab (C1-C9) thaum lub teeb pom kev tshwm sim (plaub C0) muaj lub wavelength ntawm (a) 400-750 nm (raws li qhia hauv daim duab 5a), (b) 530nm ua.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, raws.
Tus tsim cuaj-xim spectrometer tau siv rau plaub-capillary electrophoresis (kom paub meej, saib Cov Khoom Siv Ntxiv) 31,34,35.Plaub-capillary matrix muaj plaub capillaries (sab nraud txoj kab uas hla 360 μm thiab puab txoj kab uas hla 50 μm) nyob rau ntawm 1 hli intervals ntawm lub laser irradiation site.Cov qauv uas muaj DNA cov ntawv sau nrog 8 dyes, xws li 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), thiab WEN (dye 8) nyob rau hauv ascending order of fluorescent wavelength, cais nyob rau hauv txhua ntawm plaub capillaries (tom qab no hu ua Cap1, Cap2, Cap3, thiab Cap4).Laser-induced fluorescence los ntawm Cap1-Cap4 yog collimated nrog ib tug array ntawm plaub lo ntsiab muag thiab ib txhij kaw nrog ib tug cuaj-xim spectrometer.Qhov kev siv zog ntawm cuaj-xim (C1-C9) fluorescence thaum lub sij hawm electrophoresis, uas yog, cuaj-xim electrophoregram ntawm txhua lub capillary, yog qhia hauv daim duab 7a.Ib qho sib npaug cuaj-xim electrophoregram tau txais hauv Cap1-Cap4.Raws li qhia los ntawm Cap1 xub nyob rau hauv daim duab 7a, yim lub ncov ntawm txhua cuaj-xim electrophoregram qhia ib qho fluorescence emission los ntawm Dye1-Dye8, feem.
Simultaneous quantification ntawm yim dyes siv cuaj-xim plaub-capillary electrophoresis spectrometer.(a) Cuaj-xim (C1-C9) electrophoregram ntawm txhua capillary.Lub yim lub ncov qhia los ntawm cov xub Cap1 qhia tus kheej fluorescence emissions ntawm yim dyes (Dye1-Dye8).Cov xim ntawm cov xub sib raug rau cov xim (b) thiab (c).(b) Fluorescence spectra ntawm yim dyes (Dye1-Dye8) ib capillary.c Electropherograms ntawm yim dyes (Dye1-Dye8) ib capillary.Lub ncov ntawm Dye7-labeled DNA fragments yog qhia los ntawm xub, thiab lawv Cap4 lub hauv paus ntev yog qhia.
Qhov sib faib hnyav ntawm C1-C9 ntawm yim lub ncov yog qhia hauv Fig.7b ,ua.Vim tias ob qho tib si C1-C9 thiab Dye1-Dye8 nyob rau hauv qhov kev txiav txim wavelength, yim qhov kev faib tawm hauv daim duab 7b qhia cov fluorescence spectra ntawm Dye1-Dye8 raws li ntawm sab laug mus rau sab xis.Hauv txoj kev tshawb no, Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, thiab Dye8 tshwm nyob rau hauv magenta, violet, xiav, cyan, ntsuab, daj, txiv kab ntxwv, thiab liab, feem.Nco ntsoov tias cov xim ntawm cov xub hauv daim duab 7a sib haum rau cov xim xim hauv daim duab 7b.C1-C9 fluorescence intensities rau txhua spectrum nyob rau hauv daim duab 7b tau normalized kom lawv sums sib npaug ib.Yim qhov sib npaug fluorescence spectra tau txais los ntawm Cap1-Cap4.Ib tus tuaj yeem pom meej qhov sib tshooj ntawm fluorescence ntawm dye 1-dye 8.
Raws li pom nyob rau hauv daim duab 7c, rau txhua lub capillary, cuaj-xim electrophoregram nyob rau hauv daim duab 7a tau hloov mus rau ib yim-dye electropherogram los ntawm ntau yam kev tsom xam raws li yim fluorescence spectra nyob rau hauv daim duab 7b (saib Cov Khoom Ntxiv rau cov ntsiab lus).Txij li thaum lub spectral sib tshooj ntawm fluorescence nyob rau hauv daim duab 7a tsis tshwm nyob rau hauv daim duab 7c, Dye1-Dye8 tuaj yeem txheeb xyuas thiab ntsuas tus kheej ntawm txhua lub sijhawm, txawm tias ntau npaum li cas ntawm Dye1-Dye8 fluoresce tib lub sijhawm.Qhov no tsis tuaj yeem ua tiav nrog kev txheeb xyuas xya-xim 31, tab sis tuaj yeem ua tiav nrog kev tsim muaj cuaj xim.Raws li pom los ntawm cov xub Cap1 hauv daim duab 7c, tsuas yog cov fluorescent emission singlets Dye3 (xiav), Dye8 (liab), Dye5 (ntsuab), Dye4 (cyan), Dye2 (liab), Dye1 (magenta), thiab Dye6 (daj). ) raug pom nyob rau hauv qhov kev txiav txim siab chronological.Rau cov fluorescent emission ntawm dye 7 (txiv kab ntxwv), ntxiv rau ib lub ncov uas qhia los ntawm lub txiv kab ntxwv xub, ob peb lwm cov peaks tau pom.Qhov tshwm sim no yog vim qhov tseeb tias cov qauv muaj cov qauv loj, Dye7 sau npe DNA tawg nrog qhov sib txawv ntawm qhov ntev.Raws li pom nyob rau hauv daim duab 7c, rau Cap4 lub hauv paus ntev yog 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 thiab 220 puag lengths.
Cov yam ntxwv tseem ceeb ntawm cuaj-xim spectrometer, tsim los siv lub matrix ntawm ob-txheej dichroic iav, yog me me thiab tsim yooj yim.Txij li cov array ntawm decachromatic iav nyob rau hauv lub adapter qhia nyob rau hauv daim duab.3c mounted ncaj qha rau ntawm daim duab sensor board (saib daim duab S1 thiab S2), cuaj-xim spectrometer muaj tib qhov ntev ntawm lub adapter, piv txwv li 54 × 58 × 8.5 hli.(thickness).Qhov loj me me no yog ob mus rau peb qhov kev txiav txim ntawm qhov loj me dua li cov qauv spectrometers uas siv gratings lossis prisms.Tsis tas li ntawd, txij li cuaj-xim spectrometer tau teeb tsa xws li lub teeb tawm tsam qhov chaw ntawm cov duab sensor perpendicularly, qhov chaw tuaj yeem yooj yim faib rau cuaj-xim spectrometer hauv cov tshuab xws li microscopes, flow cytometers, lossis analyzers.Capillary grating electrophoresis analyzer rau ntau dua miniaturization ntawm lub cev.Nyob rau tib lub sijhawm, qhov loj ntawm kaum dichroic tsom iav thiab bandpass lim siv nyob rau hauv cuaj-xim spectrometer tsuas yog 10 × 1.9 × 0.5 hli lossis 15 × 1.9 × 0.5 hli.Yog li, ntau tshaj 100 xws li me me dichroic tsom iav thiab bandpass lim, feem, tuaj yeem txiav los ntawm daim iav dichroic thiab 60 mm2 bandpass lim, feem.Yog li ntawd, ib qho array ntawm decachromatic iav tuaj yeem tsim tau ntawm tus nqi qis.
Lwm feature ntawm cuaj-xim spectrometer yog nws cov yam ntxwv zoo spectral.Tshwj xeeb, nws tso cai rau kev nrhiav tau ntawm spectral dluab ntawm snapshots, uas yog, ib txhij nrhiav tau ntawm cov duab nrog cov ntaub ntawv spectral.Rau txhua daim duab, ib qho spectrum txuas ntxiv tau txais nrog lub wavelength ntawm 520 txog 700 nm thiab kev daws teeb meem ntawm 20 nm.Hauv lwm lo lus, cuaj xim siv lub teeb raug kuaj pom rau txhua daim duab, piv txwv li cuaj 20 nm bands sib npaug sib faib cov wavelength ntau ntawm 520 txog 700 nm.Los ntawm kev hloov cov yam ntxwv spectral ntawm daim iav dichroic thiab lub bandpass lim, lub wavelength ntau ntawm cuaj bands thiab qhov dav ntawm txhua lub band tuaj yeem hloov kho.Cuaj xim kuaj pom tuaj yeem siv tsis tau tsuas yog rau kev ntsuas fluorescence nrog spectral imaging (raws li tau piav qhia hauv daim ntawv qhia no), tab sis kuj rau ntau lwm yam kev siv siv spectral imaging.Txawm hais tias hyperspectral imaging tuaj yeem ntes ntau pua xim, nws tau pom tias txawm tias muaj qhov txo qis ntawm cov xim pom tau, ntau yam khoom hauv qhov kev pom tuaj yeem txheeb xyuas qhov tseeb txaus rau ntau daim ntawv thov38,39,40.Vim hais tias kev daws teeb meem spatial, spectral daws teeb meem, thiab kev daws teeb meem ntawm lub cev muaj kev sib pauv hauv spectral imaging, txo cov xim tuaj yeem txhim kho spatial daws teeb meem thiab kev daws teeb meem ntawm lub cev.Nws tseem tuaj yeem siv cov spectrometers yooj yim xws li ib qho tsim hauv txoj kev tshawb no thiab ntxiv txo cov nqi ntawm kev suav.
Nyob rau hauv txoj kev tshawb no, yim dyes tau muab xam ib txhij los ntawm spectral sib cais ntawm lawv overlapping fluorescence spectra raws li nrhiav tau ntawm cuaj xim.Txog cuaj dyes tuaj yeem suav tau ib txhij, sib koom ua ke hauv lub sijhawm thiab qhov chaw.Ib qho tshwj xeeb kom zoo dua ntawm cuaj-xim spectrometer yog nws lub siab luminous flux thiab loj aperture (1 × 7 mm).Lub decane daim iav array muaj qhov siab tshaj plaws kis tau tus mob ntawm 92% ntawm lub teeb los ntawm lub aperture nyob rau hauv txhua ntawm cuaj wavelength ranges.Qhov ua tau zoo ntawm kev siv teeb pom kev tshwm sim hauv lub wavelength ntau ntawm 520 txog 700 nm yog yuav luag 100%.Nyob rau hauv xws li ib tug ntau yam ntawm wavelengths, tsis muaj diffraction grating yuav muab xws li ib tug high efficiency ntawm kev siv.Txawm hais tias qhov diffraction efficiency ntawm diffraction grating ntau tshaj 90% ntawm ib qho wavelength, raws li qhov sib txawv ntawm qhov wavelength thiab ib qho wavelength nce, qhov diffraction efficiency ntawm lwm wavelength txo 41.Lub aperture dav perpendicular mus rau cov kev taw qhia ntawm lub dav hlau nyob rau hauv daim duab 2c yuav ncua ntawm 7 hli mus rau qhov dav ntawm cov duab sensor, xws li nyob rau hauv cov ntaub ntawv ntawm cov duab sensor siv nyob rau hauv txoj kev tshawb no, los ntawm me ntsis hloov cov decamer array.
Lub cuaj-xim spectrometer tuaj yeem siv tsis tau tsuas yog rau capillary electrophoresis, raws li qhia hauv txoj kev tshawb no, tab sis kuj rau ntau yam.Piv txwv li, raws li qhia hauv daim duab hauv qab no, cuaj-xim spectrometer tuaj yeem siv rau lub tshuab ntsuas fluorescence.Lub dav hlau ntawm cov qauv yog tso tawm rau ntawm daim duab sensor ntawm cuaj-xim spectrometer los ntawm 10x lub hom phiaj.Qhov kev ncua deb ntawm lub hom phiaj ntawm lub hom phiaj lens thiab cov duab sensor yog 200 hli, thaum qhov kev ncua deb ntawm qhov pom ntawm qhov teeb meem ntawm cuaj xim spectrometer thiab cov duab sensor tsuas yog 12 hli.Yog li ntawd, cov duab raug txiav mus rau kwv yees li qhov loj ntawm lub aperture (1 × 7 mm) nyob rau hauv lub dav hlau ntawm qhov xwm txheej thiab muab faib ua cuaj xim dluab.Ntawd yog, cov duab spectral ntawm cuaj xim snapshot tuaj yeem coj mus rau ntawm 0.1 × 0.7 mm cheeb tsam hauv cov qauv dav hlau.Tsis tas li ntawd, nws muaj peev xwm tau txais ib daim duab 9-xim spectral ntawm ib cheeb tsam loj ntawm cov qauv dav hlau los ntawm kev txheeb xyuas cov qauv txheeb ze rau lub hom phiaj ntawm txoj kab rov tav hauv daim duab 2c.
Lub decachromatic daim iav array Cheebtsam, uas yog M1-M9 thiab BP, tau kev cai tsim los ntawm Asahi Spectra Co., Ltd. siv cov txheej txheem nag lossis daus.Multilayer dielectric cov ntaub ntawv tau siv ib tus zuj zus mus rau kaum daim hlau quartz 60 × 60 mm loj thiab 0.5 mm tuab, ua tau raws li cov hauv qab no: M1: IA = 45 °, R ≥ 90% ntawm 520-590 nm, Tave ≥ 90% ntawm 610– 610nm ua.700 nm, M2: IA = 45 °, R ≥ 90% ntawm 520-530 nm, Tave ≥ 90% ntawm 550-600 nm, M3: IA = 45 °, R ≥ 90% ntawm 540-550 nm, Tave ≥ 90 % ntawm 570-600 nm, M4: IA = 45 °, R ≥ 90% ntawm 560-570 nm, Tave ≥ 90% ntawm 590-600 nm, M5: IA = 45 °, R ≥ 98% ntawm 580-600 nm R ≥ 98% ntawm 680-700 nm, M6: IA = 45 °, Tave ≥ 90% ntawm 600-610 nm, R ≥ 90% ntawm 630-700 nm, M7: IA = 45 °, R ≥ 90% ntawm 620-630 nm, Taw ≥ 90% ntawm 650-700 nm, M8: IA = 45 °, R ≥ 90% ntawm 640-650 nm, Taw ≥ 90% ntawm 670-700 nm, M9: IA = 45 °, R ≥ 90% ntawm 650-670 nm, Tave ≥ 90% ntawm 690-700 nm, BP: IA = 0 °, T ≤ 0.01% ntawm 505 nm, Tave ≥ 95% ntawm 530-690 nm ntawm 530 nm T ≥ 90% ntawm -690 nm thiab T ≤ 1% ntawm 725-750 nm, qhov twg IA, T, Tave, thiab R yog lub kaum sab xis ntawm qhov xwm txheej, kev xa tawm, qhov nruab nrab transmittance, thiab lub teeb pom kev tsis pom kev.
Lub teeb dawb (C0) nrog lub wavelength ntau ntawm 400-750 nm emitted los ntawm LED teeb qhov chaw (AS 3000, AS IB CORPORATION) tau collimated thiab teeb meem vertically ntawm DP ntawm ib qho array ntawm dichroic iav.Lub teeb dawb spectrum ntawm LEDs yog pom nyob rau hauv Ntxiv daim duab S3.Muab lub tank acrylic (qhov loj me 150 × 150 × 30 mm) ncaj qha rau pem hauv ntej ntawm daim iav decamera array, tawm tsam PSU.Cov pa luam yeeb tsim tawm thaum cov dej khov qhuav rau hauv dej tau muab nchuav rau hauv lub tank acrylic kom pom cuaj-xim C1-C9 cais cov kwj deg emanating los ntawm array ntawm decachromatic tsom iav.
Xwb, lub teeb dawb collimated (C0) yog dhau los ntawm lub lim ua ntej nkag mus rau DP.Cov ntxaij lim dej yog Ameslikas nruab nrab ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntawm 0.6.Tom qab ntawd siv lub tshuab lim dej (FW212C, FW212C, Thorlabs).Thaum kawg, tig lub ND lim rov qab rau.Lub bandwidths ntawm cuaj bandpass lim sib haum rau C9, C8, C7, C6, C5, C4, C3, C2 thiab C1, raws li.Lub quartz cell nrog qhov ntev ntawm 40 (optical length) x 42.5 (qhov siab) x 10 mm (dav) tau muab tso rau pem hauv ntej ntawm ib qho array ntawm decochromatic tsom iav, opposite BP.Cov pa luam yeeb tau muab los ntawm lub raj rau hauv quartz cell kom tswj cov pa luam yeeb hauv quartz cell kom pom cov cuaj-xim C1-C9 cais cov kwj tawm los ntawm daim iav decachromatic array.
Ib daim vis dis aus ntawm cuaj-xim sib cais lub teeb emanating los ntawm ib qho ntawm cov iav decanic tau ntes nyob rau hauv lub sij hawm-lapse hom ntawm iPhone XS.Khaws cov duab ntawm qhov chaw ntawm 1 fps thiab suav cov duab los tsim cov yeeb yaj kiab ntawm 30 fps (rau kev xaiv video 1) lossis 24 fps (rau cov yeeb yaj kiab yeem 2 thiab 3).
Muab 50 µm tuab stainless hlau phaj (nrog plaub 50 µm txoj kab uas hla ntawm 1 hli ntu) ntawm lub phaj diffusion.Lub teeb nrog lub wavelength ntawm 400-750 nm yog irradiated mus rau lub diffuser phaj, tau los ntawm kev hla lub teeb los ntawm lub teeb halogen los ntawm ib tug luv luv transmission lim nrog ib tug txiav tawm wavelength ntawm 700 nm.Lub teeb spectrum yog qhia nyob rau hauv Ntxiv daim duab S4.Xwb, lub teeb kuj dhau los ntawm ib qho ntawm 10 nm bandpass lim nyob nruab nrab ntawm 530, 550, 570, 590, 610, 630, 650, 670 thiab 690 nm thiab ntaus lub phaj diffuser.Raws li qhov tshwm sim, plaub lub ntsiab lus hluav taws xob nrog lub cheeb ntawm φ50 μm thiab qhov sib txawv wavelengths tau tsim rau ntawm lub phaj stainless hlau tawm tsam lub phaj diffuser.
Plaub-capillary array nrog plaub lo ntsiab muag yog mounted rau ntawm cuaj-xim spectrometer raws li qhia hauv daim duab 1 thiab 2. C1 thiab C2.Plaub lub capillaries thiab plaub lo ntsiab muag yog tib yam li hauv kev tshawb fawb yav dhau los31,34.Lub laser beam nrog lub wavelength ntawm 505 nm thiab lub zog ntawm 15 mW yog irradiated ib txhij thiab tusyees los ntawm sab mus rau lub emission ntsiab lus ntawm plaub capillaries.Cov fluorescence emissions los ntawm txhua qhov emission point yog collimated los ntawm cov coj lo ntsiab muag thiab sib cais ua cuaj xim ntws los ntawm ib tug array ntawm decachromatic iav.Qhov tshwm sim 36 ntws tau raug txhaj ncaj qha rau hauv CMOS duab sensor (C11440–52U, Hamamatsu Photonics K·K.), thiab lawv cov duab raug kaw ib txhij.
ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Applied Biosystems), 4 µl GeneScan™ 600 LIZ™ dye raug muab sib xyaw rau txhua lub capillary los ntawm kev sib xyaw 1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), 1 µl sib tov loj tus qauv.v2.0 (Thermo Fisher Scientific) thiab 14 µl dej.Lub PowerPlex® 6C Matrix Standard muaj rau DNA fragments sau nrog rau dyes: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, thiab WEN, nyob rau hauv kev txiav txim ntawm qhov siab tshaj plaws wavelength.Lub hauv paus ntev ntawm cov DNA tawg no tsis tau nthuav tawm, tab sis lub hauv paus ntev ntawm DNA cov kab lus sau nrog WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C thiab TOM-6C paub.Cov khoom sib xyaw ua ke hauv ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit muaj DNA fragment sau nrog dR6G dye.Qhov ntev ntawm lub hauv paus ntawm DNA tawg kuj tsis qhia tawm.GeneScan™ 600 LIZ™ Dye Size Standard v2.0 suav nrog 36 LIZ-labeled DNA fragments.Lub hauv paus ntev ntawm cov kab DNA no yog 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 280, 300, 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 thiab 600 puag.Cov qauv raug denatured ntawm 94 ° C rau 3 feeb, ces txias rau 5 feeb.Cov qauv raug txhaj rau hauv txhua lub capillary ntawm 26 V / cm rau 9 s thiab sib cais nyob rau hauv txhua lub capillary ntim nrog POP-7 ™ cov tshuaj polymer (Thermo Fisher Scientific) nrog qhov ntev ntawm 36 cm thiab voltage ntawm 181 V / cm thiab ib qho lub kaum sab xis ntawm 60 °.Los ntawm.
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