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Yerekana karuseli ya sisitemu eshatu icyarimwe.Koresha Utubuto na Ibikurikira kugirango unyuze mumashusho atatu icyarimwe, cyangwa ukoreshe utubuto twa slide kumpera kugirango wimuke unyuze mubice bitatu icyarimwe.
Ultra-compact (54 × 58 × 8.5 mm) hamwe na aperture (1 × 7 mm) icyenda cyamabara icyenda yakozwe, "igabanyijemo kabiri" hamwe nindorerwamo icumi zindorerwamo ebyiri, zikoreshwa mugushushanya ako kanya.Umucyo wibyabaye ufite igice cyambukiranya ntoya kurenza ubunini bwa aperture igabanijwemo umurongo uhoraho wa 20 nm ubugari na cyenda yamabara afite uburebure hagati ya 530, 550, 570, 590, 610, 630, 650, 670 na 690 nm.Amashusho yimyenda icyenda yamabara icyarimwe apimwa neza na sensor sensor.Bitandukanye nindorerwamo zisanzwe za dichroic, indorerwamo ya dichroic yateye imbere ifite ibice bibiri byihariye, ntabwo byongera umubare wamabara ashobora gupimirwa icyarimwe, ariko kandi binanoza imiterere yibishusho kuri buri mugezi wamabara.Iterambere ryamabara icyenda ikoreshwa kuri bine-capillary electrophoresis.Isesengura ryinshi icyarimwe ryamabara umunani yimukira icyarimwe muri buri capillary ukoresheje fluorescence yamabara icyenda.Kubera ko ibara icyenda ryerekana ibara ritari ultra-nto gusa kandi ridahenze, ariko kandi rifite urumuri rwinshi rwinshi kandi rukemurwa ruhagije rwerekana amashusho menshi yerekana amashusho, irashobora gukoreshwa cyane mubice bitandukanye.
Kwerekana amashusho ya Hyperspectral na multispectral byahindutse igice cyingenzi cya astronomie2, kurebera kure kwitegereza Isi3,4, kugenzura ubuziranenge bwibiryo n’amazi5,6, kubungabunga ibihangano na archeologiya7, forensics8, kubaga9, gusesengura ibinyabuzima no kwisuzumisha10,11 nibindi Umwanya wa 1 Ikoranabuhanga ryingirakamaro , 12,13.Uburyo bwo gupima urumuri rwumucyo rutangwa na buri ngingo y’ibyuka bihumanya mu rwego rwo kureba bigabanijwemo (1) gusikana ingingo (“sima”) 14,15, (2) gusikana umurongo (“panicle”) 16,17,18 , (3) uburebure bwa scan 19,20,21 na (4) amashusho22,23,24,25.Kubijyanye nubu buryo bwose, gukemura umwanya, gukemura ibintu no gukemura by'agateganyo bifitanye isano yo gucuruza 9,10,12,26.Mubyongeyeho, urumuri rusohoka rufite ingaruka zikomeye kumyumvire, ni ukuvuga igipimo cyerekana-urusaku mu mashusho yerekana amashusho26.Luminous flux, ni ukuvuga, imikorere yo gukoresha urumuri, ihwanye neza nigereranya ryumubare nyawo wapimwe wumucyo wa buri mucyo kumurongo mugihe cyumubare wuzuye wumucyo wapimye uburebure bwumurongo.Icyiciro (4) nuburyo bukwiye mugihe ubukana cyangwa urumuri rwumucyo utangwa na buri kintu cyohereza ibintu bihinduka hamwe nigihe cyangwa mugihe umwanya wa buri kintu cyohereza uhinduka uhinduka hamwe nigihe kuko urumuri rwumucyo rutangwa ningingo zose zisohora zapimwe icyarimwe.24.
Benshi muburyo bwavuzwe haruguru bwahujwe na nini nini, igoye na / cyangwa ihenze ya spekrometrike ukoresheje ibinezeza 18 cyangwa 14, 16, 22, 23 prism yo kumasomo (1), (2) na (4) cyangwa 20, 21 ya disiki ya filteri, kuyungurura amazi .Akayunguruzo ka Crystalline (LCTF) 25 cyangwa acousto-optic tunable filter (AOTF) 19 yicyiciro (3).Ibinyuranye, icyiciro (4) indorerwamo nyinshi zerekana indorerwamo ni nto kandi zidahenze bitewe nuburyo bworoshye 27,28,29,30.Byongeye kandi, bafite urumuri rwinshi cyane kuko urumuri rusangiwe na buri ndorerwamo ya dicroic (ni ukuvuga urumuri rwandujwe kandi rugaragaza urumuri rwibyabaye kuri buri ndorerwamo ya dicroic) rukoreshwa byuzuye kandi bikomeza.Nyamara, umubare wumurongo wumurongo (ni ukuvuga amabara) ugomba gupimirwa icyarimwe ugarukira kuri bine.
Amashusho ya Spectral ashingiye ku kumenya fluorescence akoreshwa muburyo bwo gusesengura ibintu byinshi mugushakisha ibinyabuzima no gusuzuma indwara 10, 13.Muri multiplexing, kubera ko analyite nyinshi (urugero, ADN cyangwa proteine zihariye) zanditseho amarangi atandukanye ya fluorescent, buri analyte igaragara kuri buri cyuka cyoherezwa mumwanya wo kureba iragereranywa hakoreshejwe isesengura ryibintu byinshi.32 isenya florescence yagaragaye yerekanwa na buri kintu cyangiza.Muri iki gikorwa, amarangi atandukanye, buri kimwe gisohora fluorescence itandukanye, irashobora gukoroniza, ni ukuvuga kubana mumwanya nigihe.Kugeza ubu, umubare ntarengwa w’amabara ashobora gushimishwa nigiti kimwe cya laser ni umunani.Iyi mipaka yo hejuru ntabwo igenwa nicyemezo cyerekana (ni ukuvuga umubare wamabara), ahubwo nubugari bwurwego rwa fluorescence (≥50 nm) nubunini bwamabara ya Stoke ihinduka (≤200 nm) kuri FRET (ukoresheje FRET) 10 .Nyamara, umubare wamabara ugomba kuba urenze cyangwa uhwanye numubare wamabara kugirango ukureho guhuzagurika kwamabara avanze 31,32.Kubwibyo, birakenewe kongera umubare wamabara apimwe icyarimwe kugeza umunani cyangwa arenga.
Vuba aha, hashyizweho ultra-compact heptachroic spectrometer (ukoresheje umurongo windorerwamo ya heptychroic hamwe na sensor ishusho yo gupima fluorcente enye).Ikirangantego ni bibiri kugeza kuri bitatu byerekana ubunini buto kuruta ibisanzwe bisanzwe ukoresheje ibishimisha cyangwa prism34,35.Ariko, biragoye gushyira indorerwamo zirenga zirindwi muri spekrometrike kandi icyarimwe ugapima amabara arenga 36,37.Hamwe no kwiyongera k'umubare w'indorerwamo za dichroic, itandukaniro ntarengwa muburebure bwinzira nziza ya optique yumucyo wa dichroic yiyongera, kandi biragoye kwerekana urumuri rwose rumuri kumurongo umwe wumva.Inzira ndende ndende ya optique yuburebure bwurumuri nabwo iriyongera, bityo ubugari bwa sprometrometre aperture (ni ukuvuga ubugari ntarengwa bwurumuri rwasesenguwe na spekrometero) buragabanuka.
Mu rwego rwo gusubiza ibibazo byavuzwe haruguru, hashyizweho ultra-compact yamabara icyenda yerekana ibara ryerekana indorerwamo ebyiri zerekana “dichroic” decachromatic mirror array hamwe na sensor ishusho yerekana amashusho ako kanya [icyiciro (4)].Ugereranije nubushakashatsi bwambere, iterambere ryerekanwe rifite itandukaniro rito muburebure bwa optique yinzira ndende kandi ntoya ntoya ya optique.Byakoreshejwe kuri electrophorei ya capillary enye kugirango hamenyekane lazeri iterwa na laser-icyenda y'amabara icyenda no kugereranya icyarimwe kwimuka icyarimwe amarangi umunani muri buri capillary.Kubera ko sprometrometeri yateye imbere ntabwo ari ultra-nto gusa kandi ihendutse, ariko kandi ifite flux nini cyane kandi ikanakemurwa bihagije kuri porogaramu nyinshi zerekana amashusho, irashobora gukoreshwa cyane mubice bitandukanye.
Imyenda gakondo y'amabara icyenda yerekanwa mugitabo.1a.Igishushanyo cyacyo gikurikira icya ultra-ntoya yabanjirije amabara arindwi ya spekrometero 31. Igizwe nindorerwamo icyenda za dicroic zitondekanye mu buryo butambitse ku nguni ya 45 ° iburyo, naho sensor ishusho (S) iri hejuru yindorerwamo icyenda.Umucyo winjira uturutse hepfo (C0) ugabanijwe nindorerwamo icyenda zindorerwamo icyenda zibiri mumucyo icyenda uzamuka (C1, C2, C3, C4, C5, C6, C7, C8 na C9).Inzira icyenda zose zamabara zigaburirwa muburyo bwa sensor kandi zimenyekana icyarimwe.Muri ubu bushakashatsi, C1, C2, C3, C4, C5, C6, C7, C8, na C9 biri murwego rwuburebure bwumuraba kandi bigereranywa na magenta, violet, ubururu, cyan, icyatsi, umuhondo, orange, umutuku-orange, na umutuku.Nubwo aya mabara akoreshwa muriyi nyandiko, nkuko bigaragara ku gishushanyo cya 3, kuko atandukanye namabara nyayo abonwa nijisho ryumuntu.
Igishushanyo mbonera cyibisanzwe kandi bishya byamabara icyenda..(b) Ibara rishya ryamabara icyenda hamwe nibice bibiri byerekana indorerwamo.Icyerekezo cyumucyo C0 kigabanyijemo ibice icyenda byamabara C1-C9 kandi bigaragazwa na sensor ishusho S.
Iterambere rishya ryamabara icyenda rifite icyerekezo cya dichroic indorerwamo ebyiri hamwe na sensor ishusho, nkuko bigaragara ku gishushanyo 1b.Mu cyiciro cyo hepfo, indorerwamo eshanu za dicroic zihengamye 45 ° iburyo, zihuza iburyo uhereye hagati rwagati rwinshi.Kurwego rwo hejuru, indorerwamo eshanu zinyongera zihengamye 45 ° ibumoso kandi ziherereye hagati ziva ibumoso.Indorerwamo yibumoso ya dicroic ya layer yo hepfo hamwe nindorerwamo ya dichroic iburyo ya layer yo hejuru iruzuzanya.Amatara yibyabaye (C0) agabanijwe kuva hepfo mo ibice bine bisohoka bya chromatic (C1-C4) nindorerwamo eshanu za dicroic iburyo hamwe na chromatic eshanu zisohoka (C5-C4) nindorerwamo eshanu za dicroic ibumoso C9).Kimwe nibisanzwe icyenda-amabara ya spekrometrike, imigezi icyenda yose yamabara yinjijwe muburyo butaziguye mumashusho (S) hanyuma bikamenyekana icyarimwe.Ugereranije Igishushanyo 1a na 1b, umuntu arashobora kubona ko mugihe cyibintu bishya byamabara icyenda yerekana ibara, byombi itandukaniro ntarengwa hamwe ninzira ndende ya optique yuburebure bwamabara icyenda.
Iyubakwa rirambuye rya ultra-ntoya-ebyiri-indorerwamo ya dichroic indorerwamo ya mm 29 (ubugari) × 31 mm (ubujyakuzimu) × 6 mm (uburebure) irerekanwa ku gishushanyo cya 2. Indorerwamo ya dicroic decimal igizwe nindorerwamo eshanu za dicroic iburyo .Indorerwamo zose za dicroic ziranyeganyezwa kugirango zishyure icyerekezo kimwe cyo kwimuka bitewe no kugabanuka kwinzira zinyuze mu ndorerwamo.Munsi ya M1, bande-pass filter (BP) irakosowe.Ibipimo bya M1 na BP ni 10mm (uruhande rurerure) x 1.9mm (uruhande rugufi) x 0.5mm (uburebure).Ibipimo by'indorerwamo zisigaye ni mm 15 × 1,9 mm × 0.5 mm.Ikibanza cya matrix hagati ya M1 na M2 ni 1,7 mm, mugihe matrike ya matrix yizindi ndorerwamo ebyiri ni mm 1,6.Ku mutini.2c ikomatanya ibyabaye kumurika C0 hamwe nicyenda cyumucyo cyamabara C1-C9, gitandukanijwe na materique de-chambre yindorerwamo.
Kubaka materix ya dichroic ibice bibiri..Igizwe nindorerwamo eshanu (M1-M5) ziri murwego rwo hasi, indorerwamo eshanu ebyiri (M6-M9 nindi M5) iherereye murwego rwo hejuru, hamwe na filteri ya bande (BP) iri munsi ya M1.(c) Kwambukiranya ibice mu cyerekezo gihagaritse, hamwe na C0 na C1-C9 byuzuye.
Ubugari bwa aperture mu cyerekezo gitambitse, cyerekanwa n'ubugari bwa C0 ku gishushanyo cya 2, c, ni mm 1, no mu cyerekezo cya perpendikulari ku ndege ya 2, c, gitangwa n'ibishushanyo mbonera bya aluminium, - mm 7.Nukuvuga, ibara rishya ryamabara icyenda rifite ubunini bunini bwa mm 1 × 7 mm.Inzira ya optique ya C4 ni ndende cyane muri C1-C9, kandi inzira ya optique ya C4 imbere mumirongo yindorerwamo ya dichroic, kubera ubunini bwa ultra-nto (29 mm × 31 mm × 6 mm), ni mm 12.Muri icyo gihe, inzira ya optique yuburebure bwa C5 niyo ngufi muri C1-C9, naho inzira ya optique ya C5 ni 5.7mm.Kubwibyo, itandukaniro ntarengwa muburebure bwa optique ni 6.3 mm.Uburebure bwa optique yinzira ikosorwa kuburebure bwinzira ya optique yo kohereza optique ya M1-M9 na BP (kuva muri quartz).
Ibiranga ibintu bya М1 - М9 na VR bibarwa ku buryo amasoko С1, С2, С3, С4, С5, С6, С7, С8 na С9 ari mu burebure bw’umuraba 520–540, 540–560, 560–580, 580 –600, 600–620, 620–640, 640–660, 660–680, na 680–700 nm.
Ifoto ya matrix yakozwe yindorerwamo ya decachromatic irerekanwa mumashusho 3a.M1-M9 na BP bifatanye kumurongo wa dogere 45 hamwe nindege itambitse ya aluminiyumu, mugihe M1 na BP byihishe inyuma yishusho.
Umusaruro windorerwamo ya decan no kwerekana.(a) Urutonde rwindorerwamo zahimbwe..(c) Urutonde rwindorerwamo ya decochromatic yamurikiwe numucyo wera uturutse inyuma..
Ibipimo byapimwe byapimwe bya M1-M9 C0 ku mpande zanduye 45 ° naho ibipimo byapimwe byapimwe bya BP C0 ku mpande zanduye 0 ° byerekanwe ku gishushanyo.4a.Ikwirakwizwa rya C1-C9 ugereranije na C0 ryerekanwe mubishushanyo.4b.Iyerekanwa ryabazwe uhereye kuri ecran.4a ukurikije inzira nziza C1-C9 mu gishushanyo cya 4a.1b na 2c.Kurugero, 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)], aho TS (X) na [1 - TS (X)] ni ihererekanyabubasha ryerekana X, kimwe.Nkuko bigaragara ku gishushanyo cya 4b, umurongo (umurongo wa ≥50%) wa C1, C2, C3, C4, C5, C6, C7, C8 na C9 ni 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 na 682-699 nm.Ibisubizo birahuye nurwego rwateye imbere.Mubyongeyeho, gukoresha neza urumuri rwa C0 ni hejuru, ni ukuvuga ko impuzandengo ntarengwa ya C1-C9 yoherejwe ni 92%.
Ikwirakwizwa ryerekana indorerwamo ya dicroic hamwe no gucamo ibara icyenda.(a) Ibipimo byapimwe byapimwe M1-M9 kuri 45 ° na BP kuri 0 °.(b) Ikwirakwizwa rya C1 - C9 ugereranije na C0 ubarwa kuva (a).
Ku mutini.3c, umurongo w'indorerwamo za dichroic uherereye mu buryo buhagaritse, ku buryo uruhande rwiburyo rwarwo ku gishushanyo cya 3a ari uruhande rwo hejuru kandi urumuri rwera rwa LED yegeranye (C0) ruri inyuma.Imirongo yindorerwamo ya decachromatic igaragara ku gishushanyo cya 3a yashyizwe muri mm 54 (uburebure) × 58 mm (ubujyakuzimu) × 8.5 mm (umubyimba) adapt.Ku mutini.3d, usibye leta yerekanwe mumitini.3c, ikigega cya acrylic cyuzuye umwotsi cyashyizwe imbere yindorerwamo zindorerwamo za decochromatic, amatara yo mucyumba yazimye.Nkigisubizo, imigezi icyenda itandukanye igaragara muri tank, ituruka kumurongo windorerwamo ya decatroic.Buri mugezi ucitsemo ibice bifite urukiramende rufite urukiramende rufite uburebure bwa mm 1 × 7 mm, bihuye nubunini bwa aperture yuburebure bushya bwamabara icyenda.Mu gishushanyo cya 3b, urupapuro rushyirwa imbere yindorerwamo yindorerwamo ya dicroic ku gishushanyo cya 3c, kandi ishusho ya mm 1 x 7 mm yimigezi icyenda ya dicroic iteganijwe ku mpapuro igaragara uhereye ku cyerekezo cyo kugenda.imigezi.Inzira icyenda zo gutandukanya imigezi mumitini.3b na d ni C4, C3, C2, C1, C5, C6, C7, C8 na C9 kuva hejuru kugeza hasi, bishobora no kugaragara mubishusho 1 na 2. 1b na 2c.Zigaragara mu mabara ahuye n'uburebure bwazo.Bitewe nubushyuhe buke bwurumuri rwa LED (reba Ishusho yinyongera. S3) hamwe nubukangurambaga bwa kamera yamabara yakoreshejwe mugufata C9 (682–699 nm) mubishusho Ibindi bitemba bitandukana.Mu buryo nk'ubwo, C9 yagaragaye neza mu jisho.Hagati aho, C2 (umugezi wa kabiri uva hejuru) isa n'icyatsi ku gishushanyo cya 3, ariko gisa n'umuhondo ku jisho.
Inzibacyuho kuva ku gishushanyo cya 3c kugeza kuri d irerekanwa muri Video yinyongera 1. Ako kanya urumuri rwera ruva kuri LED runyuze mumirongo yindorerwamo ya decachromatic, igabanyamo icyarimwe mumirongo icyenda yamabara.Mu kurangiza, umwotsi uri muri vatiri wagabanutse buhoro buhoro kuva hejuru kugeza hasi, ku buryo ifu icyenda y'amabara nayo yazimye kuva hejuru kugeza hasi.Ibinyuranye, muri Video yinyongera 2, mugihe uburebure bwumurongo wibintu bitemba urumuri kumurongo windorerwamo ya decachromatic byahinduwe kuva muburebure kugeza kuri bigufi kuri gahunda ya 690, 671, 650, 632, 610, 589, 568, 550 na 532 nm ., Gusa imigezi ijyanye ninzira icyenda yatandukanijwe ikurikiranye C9, C8, C7, C6, C5, C4, C3, C2, na C1 irerekanwa.Ikigega cya acrylic gisimburwa na pisine ya quartz, kandi flake ya buri mugezi uhindagurika irashobora kugaragara neza uhereye kumurongo ugana hejuru.Mubyongeyeho, sub-video 3 yahinduwe kuburyo igice cyo guhindura umurongo wa sub-video 2 gisubirwamo.Nibisobanuro byiza cyane biranga ibiranga decochromatic array yindorerwamo.
Ibisubizo byavuzwe haruguru byerekana ko indorerwamo ya decachromatic yakozwe cyangwa indorerwamo nshya y'amabara icyenda ikora nkuko yabigenewe.Ibara rishya ryamabara icyenda ryakozwe mugushiraho indorerwamo za decachromatic indorerwamo hamwe na adaptate kumurongo wibishusho.
Luminous flux ifite uburebure buri hagati ya 400 na 750 nm, ikoherezwa ningingo enye z'imirasire φ50 mkm, iherereye hagati ya mm 1 mu cyerekezo cya perpendicular yerekeza ku ndege ya shusho ya 2c, Ubushakashatsi 31, 34. Ubushakashatsi bwakozwe na lens enye bugizwe na lens enye φ1 mm hamwe nuburebure bwa mm 1,4 hamwe nikibanza cya mm 1.Inzira enye zegeranijwe (enye C0) ni ibyabaye kuri DP ya ecran nshya y'amabara icyenda, iri hagati ya mm 1 intera.Indorerwamo zindorerwamo ebyiri zigabanya buri mugezi (C0) mumirongo icyenda yamabara (C1-C9).Inzira 36 zavuyemo (ibice bine bya C1-C9) noneho zinjizwa mu buryo butaziguye mu cyuma cyerekana amashusho ya CMOS (S) gihujwe neza n’indorerwamo ya dicroic.Nkigisubizo, nkuko bigaragara ku gishushanyo cya 5a, bitewe ninzira ntoya nini ya optique ninzira ngufi ntarengwa ya optique, amashusho yinzuzi zose uko ari 36 yagaragaye icyarimwe kandi bigaragara neza nubunini bumwe.Ukurikije ibice byo hasi (reba Ishusho yinyongera S4), ubukana bwibishusho mumatsinda ane C1, C2 na C3 ni make.Amashusho mirongo itatu na atandatu yari 0.57 ± 0,05 mm z'ubunini (bivuze ± SD).Rero, gukuza amashusho byagereranijwe 11.4.Intera ihagaritse hagati yamashusho igereranya mm 1 (intera imwe nki lens array) naho intera itambitse igereranya mm 1,6 (intera imwe nki ndorerwamo ya dicroic).Kuberako ingano yishusho ari nto cyane kuruta intera iri hagati yamashusho, buri shusho irashobora gupimwa wigenga (hamwe ninzira ndende).Hagati aho, amashusho yinzuzi makumyabiri n'umunani zanditswe na sprometero isanzwe y'amabara arindwi yakoreshejwe mubushakashatsi bwacu bwambere irerekanwa ku gishushanyo cya 5 B. Imirongo yindorerwamo ndwi ya dicroic yakozwe mugukuraho indorerwamo ebyiri ziburyo ziburyo bwa dicroic. indorerwamo mu gishushanyo 1a.Ntabwo amashusho yose atyaye, ingano yishusho yiyongera kuva kuri C1 kugeza kuri C7.Amashusho 28 ni 0,70 ± 0.19 mm mubunini.Kubwibyo, biragoye gukomeza imiterere ihanitse mumashusho yose.Coefficient yo gutandukana (CV) kubunini bwishusho 28 mubishusho 5b yari 28%, mugihe CV kubunini bwa shusho 36 mubishusho 5a yagabanutse kugera kuri 9%.Ibisubizo byavuzwe haruguru byerekana ko ibara rishya ryamabara icyenda ritongera gusa umubare wamabara yapimwe icyarimwe kuva kuri arindwi kugeza icyenda, ariko kandi rifite ishusho ndende kuri buri bara.
Kugereranya ubuziranenge bwibishusho bitandukanijwe byakozwe nibisanzwe kandi bishya.(a) Amatsinda ane yamabara icyenda yatandukanijwe (C1-C9) yakozwe namashusho mashya y'amabara icyenda.(b) Ibice bine byamashusho arindwi yatandukanijwe (C1-C7) byakozwe hamwe nibisanzwe byamabara arindwi.Fluxes (C0) ifite uburebure bwa metero kuva kuri 400 kugeza kuri 750 nm kuva ahantu hane hasohokera hamwe hamwe nibyabaye kuri buri cyerekezo.
Ibiranga ibintu byerekana amabara icyenda yerekana ibara ryasuzumwe mu buryo bw'igeragezwa kandi ibisubizo by'isuzuma byerekanwe ku gishushanyo cya 6. Menya ko Ishusho 6a yerekana ibisubizo bimwe na Ishusho 5a, ni ukuvuga ku burebure bwa 4 C0 400–750 nm, amashusho 36 yose aramenyekana (Amatsinda 4 C1 - C9).Ibinyuranye, nkuko bigaragara ku gishushanyo cya 6b - j, iyo buri C0 ifite uburebure bwihariye bwa 530, 550, 570, 590, 610, 630, 650, 670, cyangwa 690 nm, hari amashusho ane gusa (ane) matsinda yatahuye C1, C2, C3, C4, C5, C6, C7, C8 cyangwa C9).Nyamara, amwe mumashusho yegeranye namashusho ane ahuye aramenyekana cyane kuberako C1 - C9 yoherejwe yerekanwe kumashusho ya 4b ihuzagurika gato kandi buri C0 ifite umurongo wa 10 nm kumurongo wihariye nkuko byasobanuwe muburyo.Ibisubizo birahuye na C1-C9 yoherejwe yerekanwe mubishushanyo.4b na videwo yinyongera 2 na 3. Muyandi magambo, icyenda cyamabara ya ecran ikora nkuko byari byitezwe ukurikije ibisubizo byerekanwe kumitini.4b.Kubwibyo, hanzuwe ko gukwirakwiza ubukana bwibishusho C1-C9 aribwo buryo bwa buri C0.
Ibiranga ibintu biranga ibara icyenda.Ibara rishya ryamabara icyenda ritanga ibice bine byamashusho icyenda yatandukanijwe (C1-C9) mugihe urumuri rwibyabaye (bine C0) rufite uburebure bwa (a) 400-750 nm (nkuko bigaragara ku gishushanyo 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, bikurikiranye.
Iterambere ryamabara icyenda ryakoreshejwe kuri electrophorei ya capillary enye (kubisobanuro birambuye, reba Ibikoresho byiyongera) 31,34,35.Matrix ya capillary enye igizwe na capillaries enye (diameter yo hanze 360 mkm na diametre y'imbere 50 mkm) iherereye hagati ya mm 1 kumwanya wa lazeri.Ingero zirimo ibice bya ADN byanditseho amarangi 8, aribyo FL-6C (irangi 1), JOE-6C (irangi 2), dR6G (irangi 3), TMR-6C (irangi 4), CXR-6C (irangi 5), TOM- 6C.Laser-iterwa na fluorescence kuva Cap1-Cap4 yakusanyirijwe hamwe hamwe ninzira enye kandi icyarimwe yandikwa hamwe na spekrometero y'amabara icyenda.Imbaraga zingirakamaro zamabara icyenda (C1-C9) fluorescence mugihe cya electrophoreis, ni ukuvuga amashanyarazi icyenda ya electrophoregram ya buri capillary, irerekanwa mumashusho 7a.Ihwanye n'amabara icyenda ya electrophoregram iboneka muri Cap1-Cap4.Nkuko bigaragazwa n’imyambi ya Cap1 ku gishushanyo 7a, impinga umunani kuri buri mashanyarazi icyenda yerekana amashanyarazi yerekana imyuka imwe ya fluorescence iva Dye1-Dye8.
Kugereranya icyarimwe amarangi umunani ukoresheje ibara icyenda-bine-capillary electrophoresis spectrometer.(a) Ibara icyenda (C1-C9) electrophoregram ya buri capillary.Impinga umunani zerekanwa n imyambi Cap1 yerekana imyuka ya fluorescence yumuntu ku giti cye (Dye1-Dye8).Amabara y'imyambi ahuye n'amabara (b) na (c).(b) Fluorescence yerekana amarangi umunani (Dye1-Dye8) kuri capillary.c Electropherogrammes yamabara umunani (Dye1-Dye8) kuri capillary.Impinga ya Dye7 yanditseho ibice bya ADN byerekanwa n imyambi, naho uburebure bwa Cap4 bwerekanwe.
Gukwirakwiza ubukana bwa C1 - C9 ku mpinga umunani zerekanwa mu mashusho.7b.Kuberako C1-C9 na Dye1-Dye8 byombi bikurikirana murwego rwumuraba, umunani wagabanijwe kumashusho 7b yerekana fluorescence yerekanwe ya Dye1-Dye8 ikurikiranye uhereye ibumoso ugana iburyo.Muri ubu bushakashatsi, Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, na Dye8 bigaragara muri magenta, violet, ubururu, cyan, icyatsi, umuhondo, orange, n'umutuku.Menya ko amabara yimyambi ku gishushanyo 7a ahuye namabara yamabara kumashusho 7b.Ubwinshi bwa C1-C9 fluorescence kuri buri cyerekezo ku gishushanyo 7b cyari gisanzwe kuburyo igiteranyo cyacyo kingana.Umunani uhwanye na fluorescence yerekanwe muri Cap1-Cap4.Umuntu arashobora kwitegereza neza guhuza florescence hagati y irangi 1-irangi 8.
Nkuko bigaragara ku gishushanyo cya 7c, kuri buri capillary, electrophoregram yamabara icyenda ku gishushanyo 7a yahinduwe kuri electropherogramu yamabara umunani yakozwe nisesengura ryibice byinshi ishingiye kuri spekure umunani ya fluorescence ku gishushanyo 7b (reba Ibikoresho by'inyongera kubisobanuro birambuye).Kubera ko guhuzagurika kwa fluorescence ku gishushanyo 7a bitagaragara ku gishushanyo 7c, Dye1-Dye8 irashobora kumenyekana no kugereranywa kugiti cyayo kuri buri mwanya, kabone niyo yaba atandukanye ya fluoresce ya Dye1-Dye8 icyarimwe.Ibi ntibishobora gukorwa hamwe namabara arindwi yamabara 31, ariko birashobora kugerwaho hamwe niterambere ryamabara icyenda.Nkuko bigaragazwa n’imyambi Cap1 ku gishushanyo 7c, gusa florescent isohora imyuka ya Dye3 (ubururu), Dye8 (umutuku), Dye5 (icyatsi), Dye4 (cyan), Dye2 (ibara ry'umuyugubwe), Irangi1 (magenta), na Dye6 (Umuhondo) ) byubahirizwa muburyo buteganijwe bukurikirana.Kubijyanye na fluorescent isohora irangi 7 (orange), usibye impinga imwe yerekanwa numwambi wa orange, izindi mpinga nyinshi zaragaragaye.Igisubizo giterwa nuko ingero zirimo ibipimo byubunini, Dye7 yanditseho ibice bya ADN bifite uburebure butandukanye.Nkuko bigaragara ku gishushanyo 7c, kuri Cap4 ubu burebure bwibanze ni 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 na 220 uburebure bwibanze.
Ibintu nyamukuru biranga amabara icyenda yerekana amabara, yatejwe imbere akoresheje matrix yindorerwamo ebyiri-indorerwamo ebyiri, ni ntoya kandi yoroheje.Kuva umurongo windorerwamo ya decachromatic imbere muri adaptor yerekanwe mumitini.3c yashyizwe ku kibaho cyerekana amashusho (reba Ishusho ya S1 na S2), ibara ryerekana icyenda rifite ibipimo bimwe na adapt, ni ukuvuga 54 × 58 × 8.5 mm.(ubunini).Ingano ya ultra-nto ni bibiri kugeza kuri bitatu byerekana ubunini buto kuruta ibisanzwe bisanzwe bikoresha ibishimisha cyangwa prism.Mubyongeyeho, kubera ko ibara icyenda ryerekana ibara ryerekanwe kuburyo urumuri rukubita hejuru ya sensor ya perpendicularly, umwanya urashobora kugenerwa byoroshye kuri cyenda y'amabara icyenda muri sisitemu nka microscopes, cytometero itemba, cyangwa abasesengura.Capillary grating electrophoresis isesengura kugirango irusheho kuba miniaturizasi ya sisitemu.Muri icyo gihe, ubunini bw'indorerwamo icumi za dicroic na bande zungurura zikoreshwa muri cyenda y'amabara icyenda ni 10 × 1.9 × 0.5 mm cyangwa 15 × 1.9 × 0.5 mm.Rero, indorerwamo zirenga 100 ntoya nindorerwamo ntoya hamwe na filteri ya bande, birashobora gucibwa mumirorerwamo ya dicroic na 60 mm2 ya bande ya filteri.Kubwibyo, umurongo windorerwamo ya decachromatic urashobora gukorwa ku giciro gito.
Ikindi kintu kiranga amabara icyenda yerekana ibintu byiza cyane biranga ibintu.By'umwihariko, yemerera kubona amashusho yerekana amashusho, ni ukuvuga, icyarimwe kubona amashusho hamwe namakuru yerekana.Kuri buri shusho, umurongo uhoraho wabonetse hamwe nuburebure bwumuraba kuva kuri 520 kugeza kuri 700 nm no gukemura 20 nm.Muyandi magambo, icyenda cyimbaraga zumucyo zigaragara kuri buri shusho, ni ukuvuga imirongo icyenda 20 nm igabanya kimwe uburebure bwa 520 kugeza 700 nm.Muguhindura ibiranga indorerwamo ya dichroic hamwe na bande ya filteri, uburebure bwumurongo wa bande icyenda nubugari bwa buri bande birashobora guhinduka.Kugaragaza amabara icyenda ntibishobora gukoreshwa gusa mubipimo bya fluorescence hamwe no kwerekana amashusho (nkuko byasobanuwe muri iyi raporo), ariko no mubindi bikorwa byinshi bisanzwe ukoresheje amashusho yerekana.Nubwo amashusho ya hyperspectral ashobora kumenya amabara amagana, byagaragaye ko nubwo hagabanutse cyane umubare wamabara agaragara, ibintu byinshi murwego rwo kureba birashobora kumenyekana hamwe nukuri bihagije kubikorwa byinshi 38,39,40.Kuberako imiterere yikibanza, imiterere yikurikiranabikorwa, hamwe nicyemezo cyigihe gito bifite aho bihurira no kwerekana amashusho, kugabanya umubare wamabara birashobora kunoza imiterere yikibanza no gukemura byigihe gito.Irashobora kandi gukoresha spekrometrike yoroshye nkiyateye imbere muri ubu bushakashatsi kandi ikagabanya umubare wibara.
Muri ubu bushakashatsi, amarangi umunani yagereranijwe icyarimwe no gutandukanya ibintu bitandukanye bya fluorescence yerekanwe hejuru yo kumenya amabara icyenda.Amabara agera kuri icyenda arashobora kubarwa icyarimwe, kubana mugihe n'umwanya.Inyungu idasanzwe yamabara icyenda yerekana ibara ryinshi ni flux yamurika kandi nini nini (1 × 7 mm).Indorerwamo ya decane ifite ihererekanyabubasha rya 92% yumucyo uva kuri aperture muri buri cyenda cyenda.Ubushobozi bwo gukoresha urumuri rwibyabaye muburebure bwa 520 kugeza 700 nm hafi 100%.Muburyo bunini bwuburebure bwumurongo, nta gutandukanya gushobora gutanga uburyo bwiza bwo gukoresha.Nubwo itandukanyirizo ryimikorere ya diffaction ya grading irenga 90% muburebure bwumurongo runaka, nkuko itandukaniro riri hagati yuburebure bwumurongo nuburebure bwumurongo wiyongera, imikorere yo gutandukana kurindi zuba iragabanuka41.Ubugari bwa aperture perpendicular yerekeza ku cyerekezo cyindege ku gishushanyo cya 2c irashobora kwagurwa kuva kuri mm 7 kugeza ku bugari bw’imashini yerekana amashusho, nko mu gihe cya sensor ishusho yakoreshejwe muri ubu bushakashatsi, uhinduye gato icyerekezo cya decamer.
Amabara icyenda yerekana amabara ntashobora gukoreshwa gusa kuri capillary electrophoresis, nkuko bigaragara muri ubu bushakashatsi, ariko no mubindi bikorwa bitandukanye.Kurugero, nkuko bigaragara ku gishushanyo gikurikira, ibara ryerekana icyenda rishobora gukoreshwa kuri microscope ya fluorescence.Indege yicyitegererezo irerekanwa kumashusho yerekana amashusho icyenda y'amabara yerekanwe binyuze muri 10x.Intera ya optique hagati yintego yibikoresho hamwe na sensor ishusho ni mm 200, mugihe intera ihanitse hagati yibyabaye hejuru yicyerekezo cyamabara icyenda na sensor ya mashusho ni mm 12 gusa.Kubwibyo, ishusho yaciwe hafi yubunini bwa aperture (1 × 7 mm) mu ndege yibyabaye hanyuma igabanywamo amashusho icyenda y'amabara.Nukuvuga, ishusho yerekana amashusho icyenda yerekana ifoto irashobora gufatwa kumwanya wa 0.1 × 0.7 mm mu ndege ntangarugero.Mubyongeyeho, birashoboka kubona ishusho icyenda y'amabara yerekana ahantu hanini ku ndege y'icyitegererezo mugusuzuma icyitegererezo ugereranije n'intego mu cyerekezo gitambitse ku gishushanyo cya 2c.
Indorerwamo ya decachromatic igizwe nibikoresho, aribyo M1-M9 na BP, byakozwe na Asahi Spectra Co., Ltd. hakoreshejwe uburyo busanzwe bwimvura.Ibikoresho byinshi bya dielectric byakoreshwaga kugiti cya plaque ya quartz icumi mm 60 × 60 mm z'ubunini na 0,5 mm z'ubugari, byujuje ibi bikurikira: M1: IA = 45 °, R ≥ 90% kuri 520-590 nm, Tave ≥ 90% kuri 610– 610 nm.700 nm, M2: IA = 45 °, R ≥ 90% kuri 520-530 nm, Tave ≥ 90% kuri 550-600 nm, M3: IA = 45 °, R ≥ 90% kuri 540-550 nm, Tave ≥ 90 % kuri 570–600 nm, M4: IA = 45 °, R ≥ 90% kuri 560-570 nm, Tave ≥ 90% kuri 590-600 nm, M5: IA = 45 °, R ≥ 98% kuri 580-600 nm , R ≥ 98% kuri 680–700 nm, M6: IA = 45 °, Tave ≥ 90% kuri 600-610 nm, R ≥ 90% kuri 630–700 nm, M7: IA = 45 °, R ≥ 90% kuri 620–630 nm, Taw ≥ 90% kuri 650–700 nm, M8: IA = 45 °, R ≥ 90% kuri 640–650 nm, Taw ≥ 90% kuri 670–700 nm, M9: IA = 45 °, R ≥ 90% kuri 650-670 nm, Tave ≥ 90% kuri 690-700 nm, BP: IA = 0 °, T ≤ 0.01% kuri 505 nm, Tave ≥ 95% kuri 530-690 nm kuri 530 nm T ≥ 90% kuri -690 nm na T ≤ 1% kuri 725-750 nm, aho IA, T, Tave, na R arizo mpande zanduye, kwanduza, impuzandengo yikigereranyo, hamwe n’umucyo utagaragara.
Itara ryera (C0) rifite uburebure bwa metero 400-750 nm zasohowe nisoko yumucyo LED (AS 3000, NKUMWE CORPORATION) yegeranijwe kandi ibaye ihagaritse kuri DP yindorerwamo zitandukanye.Umucyo wera urumuri rwa LED rwerekanwe mugice cya S3.Shira ikigega cya acrylic (ibipimo 150 × 150 × 30 mm) imbere yimbere yindorerwamo ya decamera, ahateganye na PSU.Umwotsi wabyaye igihe urubura rwumye rwinjijwe mumazi noneho rusukwa muri tank ya acrylic kugirango turebe imigezi icyenda ya C1-C9 yacitsemo ibice biva mu ndorerwamo zindorerwamo.
Ubundi, urumuri rwera rwegeranijwe (C0) rwanyujijwe muyungurura mbere yo kwinjira muri DP.Muyunguruzi mu ntangiriro byari bitagira aho bibogamiye muyunguruzi bifite ubucucike bwa 0.6.Noneho koresha akayunguruzo ka moteri (FW212C, FW212C, Thorlabs).Hanyuma, hindura ND muyunguruzi.Umuyoboro mugari wa cyenda ya bande ya filteri ihuye na C9, C8, C7, C6, C5, C4, C3, C2 na C1.Akagari ka Quartz gafite ubunini bwimbere bwa 40 (uburebure bwa optique) x 42.5 (uburebure) x 10 mm (ubugari) bwashyizwe imbere yindorerwamo yindorerwamo ya decochromatic, ahateganye na BP.Umwotsi uhita ugaburirwa unyuze mu muyoboro wa selire ya quartz kugirango ukomeze kwibanda ku mwotsi mu kagari ka quartz kugira ngo ugaragaze imigezi icyenda ya C1-C9 yatandukanijwe iva mu ndorerwamo ya decachromatic.
Amashusho yerekana amabara icyenda yatandukanijwe yumucyo uturuka kumurongo windorerwamo ya decanic yafashwe muburyo butinda kuri iPhone XS.Fata amashusho yerekana kuri 1 fps hanyuma ukusanye amashusho kugirango ukore amashusho kuri 30 fps (kuri videwo itabishaka 1) cyangwa 24 fps (kuri videwo 2 na 3).
Shyira icyuma cya 50 µm cyuma kitagira umuyonga (hamwe na bine ya 50 µm ya diameter ya mm 1 intera) kuri plaque.Umucyo ufite uburebure bwa 400-750 nm urabagirana ku isahani ya diffuzeri, uboneka binyuze mu gucana urumuri ruva mu itara rya halogene unyuze muyungurura rugufi rufite uburebure bwa 700 nm.Umucyo urumuri rwerekanwe mugice cya S4.Ubundi, urumuri narwo runyura muri imwe muri 10 nm ya bande ya filteri ya centre hagati ya 530, 550, 570, 590, 610, 630, 650, 670 na 690 nm ikubita icyapa cya diffuzeri.Kubera iyo mpamvu, ingingo enye zerekana imirasire ifite umurambararo wa φ50 mm hamwe nuburebure butandukanye bw’umuraba byakozwe ku isahani idafite ingese ihagaze ku isahani ya diffuzeri.
Imirongo ine ya capillary ifite lens enye yashyizwe kuri cyenda y'amabara icyenda nkuko bigaragara ku gishushanyo 1 na 2. C1 na C2.Capillaries enye ninzira enye byari bimwe nkubushakashatsi bwabanje31,34.Urumuri rwa lazeri rufite uburebure bwa 505 nm nimbaraga za mW 15 zirabagirana icyarimwe kandi zingana kuva kuruhande kugeza aho imyuka ihumeka ya capillaries enye.Fluorescence itangwa na buri cyuka cyanduye ikusanyirizwa hamwe ninzira ihuye hanyuma igatandukanywa mumigezi icyenda yamabara hamwe nindorerwamo ya decachromatic.Inzira 36 zavuyemo zahise zinjizwa mu buryo butaziguye mu mashusho yerekana amashusho ya CMOS (C11440–52U, Hamamatsu Photonics K · K.), Amashusho yabo yandikwa icyarimwe.
ଏବv2.0 (Thermo Fisher Scientific) na 14 µl y'amazi.Ububasha bwa PowerPlex® 6C Matrix bugizwe nibice bitandatu bya ADN byanditseho amarangi atandatu: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, na WEN, ukurikije uburebure ntarengwa.Uburebure bwibanze bwibice bya ADN ntibwashyizwe ahagaragara, ariko uburebure bwibanze bwibanze bwibice bya ADN byanditseho WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C na TOM-6C birazwi.Uruvange muri ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit irimo igice cya ADN cyanditseho irangi rya dR6G.Uburebure bwibanze bwibice bya ADN nabyo ntibitangazwa.GeneScan ™ 600 LIZ ™ Ingano Irangi Ibipimo v2.0 ikubiyemo ibice 36 bya LIZ byanditseho ADN.Uburebure bwibanze bwibice bya ADN ni 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 300, 314, 320, 340, 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 na 600 shingiro.Ingero zerekanwe kuri 94 ° C muminota 3, hanyuma zikonjeshwa kurubura muminota 5.Ingero zatewe muri buri capillary kuri 26 V / cm kuri 9 s hanyuma zigatandukana muri buri capillary yuzuyemo igisubizo cya POP-7 ™ polymer (Thermo Fisher Scientific) gifite uburebure bwa cm 36 na voltage ya 181 V / cm na an inguni ya 60 °.KUVA.
Amakuru yose yabonetse cyangwa yasesenguwe mugihe cyubu bushakashatsi yashyizwe muriyi ngingo yatangajwe namakuru yinyongera.Andi makuru ajyanye nubu bushakashatsi araboneka kubanditsi babisabwe babisabye.
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