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Yana nuna carousel na nunin faifai uku lokaci guda.Yi amfani da maɓallan da suka gabata da na gaba don matsawa ta cikin nunin faifai guda uku a lokaci ɗaya, ko amfani da maɓallan maɓalli a ƙarshen don matsawa ta cikin nunin faifai uku a lokaci ɗaya.
An ƙirƙira wani ƙaramin ƙaramin ƙarfi (54 × 58 × 8.5 mm) da faɗin buɗe ido (1 × 7 mm) na'urar sikirin launi tara, “raba biyu” ta tsarar madubin dichroic guda goma, waɗanda aka yi amfani da su don hoton gani nan take.Juyin hasken da ya faru tare da sashin giciye ƙasa da girman buɗewa an raba shi zuwa ci gaba da tsiri mai faɗin 20 nm da ɗumbin launuka tara tare da tsayin raƙuman ruwa na 530, 550, 570, 590, 610, 630, 650, 670 da 690 nm.Hotunan rafukan launi guda tara ana auna su cikin inganci lokaci guda ta hanyar firikwensin hoto.Ba kamar nau'in madubi na dichroic na al'ada ba, ƙirar madubi na dichroic da aka haɓaka yana da ƙayyadaddun ƙayyadaddun nau'i biyu, wanda ba wai kawai yana ƙara yawan launi da za a iya aunawa a lokaci ɗaya ba, amma kuma yana inganta ƙudurin hoto ga kowane rafi mai launi.Ana amfani da spectrometer mai launi tara da aka haɓaka don electrophoresis mai capillary hudu.Binciken ƙididdiga na lokaci ɗaya na rinayen guda takwas suna ƙaura lokaci guda a cikin kowace capillary ta amfani da hasken wuta mai launin Laser mai launi tara.Tun da na'urar sikirin launi tara ba ƙaramin ƙarami ba ce kuma mara tsada, amma kuma tana da ɗimbin haske mai haske da isasshiyar ƙudiri don mafi yawan aikace-aikacen hoto, ana iya amfani da shi sosai a fagage daban-daban.
Hoto mai zurfi da yawa ya zama wani muhimmin sashi na astronomy2, nesa nesa don kallon duniya3,4, sarrafa ingancin abinci da ruwa5,6, adana fasaha da ilimin kimiya na kayan tarihi7, forensics8, tiyata9, nazarin halittu da bincike-biyu10,11 da dai sauransu. Filin 1 Fasaha mai mahimmanci. ,12,13.Hanyoyi don auna bakan hasken da ke fitarwa ta kowace wurin da ke fitowa a fagen kallo sun kasu zuwa (1) duban maki (“tsintsiya”)14,15, (2) linzamin kwamfuta (“panicle”)16,17,18 , (3) tsayin sikanin raƙuman ruwa19,20,21 da (4) hotuna22,23,24,25.A cikin yanayin duk waɗannan hanyoyin, ƙudurin sararin samaniya, ƙudirin gani da ƙudurin ɗan lokaci suna da alaƙar ciniki9,10,12,26.Bugu da ƙari, fitowar haske yana da tasiri mai mahimmanci akan hankali, watau siginar-zuwa-amo rabo a cikin hoto na gani26.Haske mai haske, wato, ingancin amfani da haske, yana daidaita kai tsaye da ma'aunin ma'aunin ma'aunin haske na kowane ma'aunin haske a kowane lokacin raka'a zuwa jimlar adadin hasken da aka auna kewayon tsayin raƙuman ruwa.Nau'i (4) hanya ce da ta dace a lokacin da ƙarfi ko bakan hasken da kowane wurin fitarwa ya canza tare da lokaci ko kuma lokacin da kowane matsayi na kowane wuri ya canza tare da lokaci saboda ana auna bakan hasken da ke fitarwa ta kowane wuri guda ɗaya.24.
Yawancin hanyoyin da ke sama ana haɗe su tare da manyan, hadaddun da / ko masu tsada masu tsada ta amfani da 18 gratings ko 14, 16, 22, 23 prisms don azuzuwan (1), (2) da (4) ko 20, 21 tace diski, masu tace ruwa. .Crystalline Tunable filters (LCTF)25 ko acousto-optic tunable filters (AOTF)19 na rukuni (3).Sabanin haka, nau'in (4) na'urorin kallon madubi masu yawa ƙanana ne kuma ba su da tsada saboda sauƙin daidaita su27,28,29,30.Bugu da ƙari, suna da babban haske mai haske saboda hasken da kowane madubi dichroic ya raba (wato, watsawa da haske na hasken abin da ya faru akan kowane madubi dichroic) cikakke ne kuma a ci gaba da amfani da shi.Koyaya, adadin madaukai masu tsayi (watau launuka) waɗanda dole ne a auna su lokaci guda yana iyakance zuwa kusan huɗu.
Ana amfani da Hotuna na Spectral bisa ga ganewar haske mai haske don nazarin multix a cikin binciken kwayoyin halitta da bincike 10, 13.A cikin ɗimbin yawa, tunda analytes da yawa (misali, takamaiman DNA ko sunadaran sunadaran) ana lakafta su tare da rini mai kyalli daban-daban, kowane manazarci da ke cikin kowane wurin fitarwa a fagen ra'ayi ana ƙididdige su ta amfani da bincike mai yawa.32 yana rushe bakan mai walƙiya da aka gano ta kowace wurin fitarwa.A lokacin wannan tsari, rini daban-daban, kowannensu yana fitar da haske daban-daban, na iya rikidewa, wato, zama tare a sararin samaniya da lokaci.A halin yanzu, matsakaicin adadin rinannun da za a iya jin daɗi ta hanyar katako na Laser guda takwas shine takwas33.Wannan babban iyaka ba a ƙayyade shi ta hanyar ƙudirin gani ba (watau adadin launuka), amma ta faɗin bakan fluorescence (≥50 nm) da adadin canza launin rini (≤200 nm) a FRET (ta amfani da FRET)10 .Koyaya, adadin launuka dole ne ya fi girma ko daidai da adadin rini don kawar da ruɗewar rini mai gauraye31,32.Sabili da haka, ya zama dole don ƙara adadin launuka masu auna lokaci guda zuwa takwas ko fiye.
Kwanan nan, an ɓullo da wani matsananci-compact heptachroic spectrometer (ta yin amfani da tsararrun madubin heptychroic da na'urar firikwensin hoto don auna filaye huɗu).Na'urar sikirin ita ce umarni biyu zuwa uku na girman girma fiye da na'urar gani ta al'ada ta amfani da gratings ko prisms34,35.Koyaya, yana da wahala a sanya madubai sama da bakwai dichroic a cikin na'urar kallo kuma a lokaci guda auna sama da launuka bakwai36,37.Tare da karuwa a cikin adadin madubai na dichroic, matsakaicin bambanci a cikin tsayin hanyoyi na hanyoyi na hasken wuta na dichroic yana ƙaruwa, kuma yana da wuya a nuna duk hasken wuta a kan jirgin sama mai hankali.Mafi tsayin hanyar gani mafi tsayi na jujin haske shima yana ƙaruwa, don haka nisa na buɗewar spectrometer (watau matsakaicin faɗin hasken da na'urar gani ta bincika) yana raguwa.
Dangane da matsalolin da ke sama, an ɓullo da ƙwaƙƙwaran ƙaƙƙarfan ƙaƙƙarfan spectrometer mai launi tara tare da tsararrun madubi na decachromatic "dichroic" Layer biyu da kuma firikwensin hoto don hoton gani nan take [class (4)].Idan aka kwatanta da na'urorin da suka gabata, na'urar sikirin da aka haɓaka tana da ɗan ƙaramin bambanci a matsakaicin tsayin hanyar gani da ƙarami matsakaicin tsayin hanyar gani.An yi amfani da shi zuwa electrophoresis mai capillary hudu don gano hasken haske mai launi tara na Laser da kuma ƙididdige ƙaura na rina takwas a cikin kowane capillary.Tunda na'urar sikirin da aka haɓaka ba ƙaramin ƙarami ba ce kuma mara tsada, amma kuma tana da babban juyi mai haske da isasshiyar ƙudurin gani don mafi yawan aikace-aikacen hoto, ana iya amfani da shi sosai a fagage daban-daban.
Ana nuna ma'aunin sikeli mai launi tara na gargajiya a cikin fig.1 a.Zanensa ya biyo bayan na baya ultra-kananan kalamai bakwai spectrometer 31. Ya ƙunshi madubin dichroic guda tara da aka shirya a kwance a kusurwar 45 ° zuwa dama, kuma firikwensin hoton (S) yana saman madubin dichroic tara.Hasken da ke shigowa daga ƙasa (C0) yana rarraba ta hanyar tsararrun madubai dichroic guda tara zuwa haske guda tara masu gudana (C1, C2, C3, C4, C5, C6, C7, C8 da C9).Dukkan rafukan launi guda tara ana ciyar da su kai tsaye zuwa firikwensin hoton kuma ana gano su a lokaci guda.A cikin wannan binciken, C1, C2, C3, C4, C5, C6, C7, C8, da C9 suna cikin tsari na tsawon tsayi kuma ana wakilta su da magenta, violet, blue, cyan, kore, rawaya, orange, ja-orange, da kuma wakilta. ja, bi da bi.Ko da yake ana amfani da waɗannan zane-zanen launi a cikin wannan takarda, kamar yadda aka nuna a hoto na 3, saboda sun bambanta da ainihin launukan da idon ɗan adam ke gani.
Zane-zane na al'ada da sabbin na'urori masu launi tara.(a) Na al'ada spectrometer mai launi tara tare da tsararrun madubai dichroic tara.(b) Sabbin spectrometer mai launi tara tare da tsarar madubin dichroic mai Layer biyu.C0 na hasken da ya faru ya kasu kashi tara masu launi C1-C9 kuma firikwensin hoton S ya gano shi.
Sabuwar spectrometer mai launi tara da aka haɓaka tana da grating dichroic madubi mai Layer biyu da firikwensin hoto, kamar yadda aka nuna a hoto 1b.A cikin ƙananan bene, madubin dichroic guda biyar suna karkatar da 45° zuwa dama, suna daidaitawa zuwa dama daga tsakiyar tsararrun decamers.A saman matakin, ƙarin ƙarin madubin dichroic guda biyar suna karkatar da 45° zuwa hagu kuma suna samuwa daga tsakiya zuwa hagu.Madubin dichroic na hagu na ƙasan Layer da madubin dichroic na dama na saman Layer sun mamaye juna.Hasken hasken da ya faru (C0) ya kasu daga ƙasa zuwa huɗu masu fita chromatic fluxes (C1-C4) ta madubin dichroic guda biyar a dama da fiɗaɗɗen chromatic fluxes biyar (C5-C4) ta madubin dichroic biyar a hagu C9).Kamar na'urori masu launi tara na al'ada, duk rafukan launi tara ana allura kai tsaye cikin firikwensin hoton (S) kuma ana gano su a lokaci guda.Kwatanta Figures 1a da 1b, mutum zai iya ganin cewa a cikin yanayin sabon nau'in nau'in nau'in nau'in launi guda tara, duka matsakaicin bambanci da tsayin hanya mafi tsayi na sauye-sauyen launi guda tara sun ragu.
An nuna cikakken cikakken ginin madaidaicin ƙaramin Layer dichroic madubi tsararrun madubi 29 mm (nisa) × 31 mm (zurfin) × 6 mm (tsawo) a cikin Hoto 2. Tsarin madubi na dichroic na decimal ya ƙunshi madubai dichroic guda biyar a dama. (M1-M5) da madubin dichroic guda biyar a gefen hagu (M6-M9 da wani M5), kowane madubi dichroic yana daidaitawa a cikin madaidaicin aluminum.Duk madubin dichroic suna jujjuyawa don ramawa daidai gwargwado saboda karkatar da kwararar da ke cikin madubin.A ƙasa M1, an gyara matattarar band-pass (BP).Girman M1 da BP sune 10mm (gefe mai tsayi) x 1.9mm (gajeren gajere) x 0.5mm (kauri).Girman sauran madubin dichroic sune 15 mm × 1.9 mm × 0.5 mm.Matsayin matrix tsakanin M1 da M2 shine 1.7 mm, yayin da matrix filin sauran madubin dichroic shine 1.6 mm.A kan fig.2c ya haɗu da saurin haske na C0 da ya faru da hasken wuta masu launi tara C1-C9, wanda aka raba da matrix de-chamber na madubai.
Gina matrix dichroic madubi mai Layer biyu.(a) Ra'ayin hangen nesa da (b) ra'ayi na giciye na tsararrun madubi dichroic mai Layer biyu (girman 29 mm x 31 mm x 6 mm).Ya ƙunshi madubin dichroic guda biyar (M1-M5) waɗanda ke cikin ƙananan Layer, madubai dichroic guda biyar (M6-M9 da wani M5) waɗanda ke cikin Layer na sama, da kuma matattarar bandpass (BP) da ke ƙasa da M1.(c) Ra'ayin ƙetare a madaidaiciyar hanya, tare da C0 da C1-C9 sun zoba.
Nisa na budewa a cikin madaidaiciyar hanya, wanda aka nuna ta hanyar nisa C0 a cikin siffa 2, c, shine 1 mm, kuma a cikin hanyar da ta dace da jirgin sama na 2, c, wanda aka ba da shi ta hanyar ƙirar aluminum. - 7 mm.Wato sabon na'ura mai launi tara yana da babban girman buɗaɗɗen 1 mm × 7 mm.Hanya na gani na C4 ita ce mafi tsayi tsakanin C1-C9, kuma hanyar gani na C4 a cikin tsararrun madubi na dichroic, saboda girman ƙananan ƙananan (29 mm × 31 mm × 6 mm), shine 12 mm.A lokaci guda, tsayin hanyar hanyar gani na C5 shine mafi guntu tsakanin C1-C9, kuma tsawon hanyar gani na C5 shine 5.7mm.Saboda haka, matsakaicin bambanci a cikin tsawon hanya na gani shine 6.3 mm.Ana gyara tsayin hanyoyin da ke sama don tsayin hanya na gani don watsawar gani na M1-M9 da BP (daga ma'adini).
Ana ƙididdige kaddarorin na М1-М9 da VR ta yadda zazzagewar С1, С2, С3, С4, С5, С6, С7, С8 da С9 suna cikin kewayon tsayin 520-540, 540-560, 560-8080, -600, 600-620, 620-640, 640-660, 660-680, da 680-700 nm, bi da bi.
Ana nuna hoton matrix ɗin da aka kera na madubin decachromatic a cikin siffa 3a.M1-M9 da BP suna manne zuwa gangaren 45-digiri da jirgin sama a kwance na tallafin aluminum, bi da bi, yayin da M1 da BP ke ɓoye a baya na adadi.
Samar da tsararrun madubin decan da nunin sa.(a) Tsari na madubin decachromatic ƙirƙira.(b) Hoton tsagaggen launi 1 mm × 7 mm tara wanda aka zayyana akan takardar da aka sanya a gaban ɗimbin madubin decachromatic da haske mai haske tare da farin haske.(c) Tsare-tsare na madubai na decochromatic wanda aka haskaka da farin haske daga baya.(d) rafi mai tsaga launi tara da ke fitowa daga jeri na madubi, ana lura da shi ta hanyar ajiye kwandon acrylic mai hayaƙi a gaban tsararrun madubin da aka lalatar a c da duhun ɗakin.
Ma'auni na watsawa na M1-M9 C0 a kusurwar abin da ya faru na 45 ° da ma'auni na watsawa na BP C0 a kusurwar abin da ya faru na 0 ° an nuna su a cikin Figs.4a ba.Ana nuna sifofin watsawa na C1-C9 dangane da C0 a cikin Fig.4b ku.An ƙididdige waɗannan bakan daga bakan da ke cikin Figs.4a daidai da hanyar gani na C1-C9 a cikin siffa 4a.1b da 2c.Misali, 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)], inda TS (X) da [1 - TS (X)] su ne watsawa da baƙaƙen tunani na X, bi da bi.Kamar yadda aka nuna a cikin Hoto 4b, bandwidths (bandwidth ≥50%) na C1, C2, C3, C4, C5, C6, C7, C8 da C9 sune 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 da 682-699 nm.Waɗannan sakamakon sun yi daidai da kewayon da aka haɓaka.Bugu da ƙari, ingancin amfani da hasken C0 yana da girma, wato, matsakaicin matsakaicin matsakaicin hasken C1-C9 shine 92%.
Siffar watsawa na madubi dichroic da rarrabuwar launuka tara.(a) Ma'auni na watsawa na M1-M9 a 45 ° abin da ya faru da BP a 0 ° abin da ya faru.(b) Bakan watsawa na C1-C9 dangane da C0 da aka lasafta daga (a).
A kan fig.3c, tsararrun madubai na dichroic suna tsaye a tsaye, ta yadda gefen dama a cikin siffa 3a shine gefen sama kuma farar katako na LED collimated (C0) yana da baya.Tsare-tsare na madubin decachromatic da aka nuna a cikin Hoto 3a an ɗora su a cikin adaftan 54 mm (tsawo) × 58 mm (zurfin) × 8.5 mm (kauri).A kan fig.3d, ban da jihar da aka nuna a fig.3c, an sanya tankin acrylic mai cike da hayaki a gaban tsararrun madubai na decochromatic, tare da kashe fitulun dakin.A sakamakon haka, rafukan dichroic guda tara suna iya gani a cikin tanki, suna fitowa daga tsararrun madubai na decatroic.Kowane rafi mai tsaga yana da sashin giciye na rectangular tare da girma na 1 × 7 mm, wanda yayi daidai da girman buɗaɗɗen sabbin sifofi masu launi tara.A cikin hoto na 3b, an sanya takardar takarda a gaban tsararrun madubai na dichroic a cikin Hoto na 3c, kuma an lura da hoton 1 x 7 mm na rafukan dichroic tara da aka zana a kan takarda daga jagorancin takarda.magudanan ruwa.Rabewar launi guda tara a cikin fig.3b da d sune C4, C3, C2, C1, C5, C6, C7, C8 da C9 daga sama zuwa kasa, wanda kuma ana iya gani a cikin adadi na 1 da 2. 1b da 2c.Ana lura da su cikin launuka masu dacewa da tsayin igiyoyinsu.Saboda ƙarancin hasken farin haske na LED (duba Ƙarin Hoto S3) da kuma ji na kyamarar launi da aka yi amfani da ita don ɗaukar C9 (682-699 nm) a cikin siffa. Sauran raƙuman ruwa suna da rauni.Hakazalika, C9 ya kasance a suma a bayyane ga ido tsirara.A halin yanzu, C2 (rafi na biyu daga sama) yayi kama da kore a cikin Hoto 3, amma ya fi launin rawaya zuwa ido tsirara.
Ana nuna sauyawa daga Hoto 3c zuwa d a Ƙarin Bidiyo 1. Nan da nan bayan farin haske daga LED ya wuce ta cikin tsararrun madubi na decachromatic, ya rabu lokaci guda zuwa kogunan launi tara.A ƙarshe, hayaƙin da ke cikin vat a hankali ya bace daga sama zuwa ƙasa, don haka foda masu launi tara suma sun ɓace daga sama zuwa ƙasa.Sabanin haka, a cikin Ƙarin Bidiyo 2, lokacin da aka canza tsayin tsayin abin da ya faru na jujjuyar haske a kan tsararrun madubin decachromatic daga dogon lokaci zuwa gajere a cikin tsari na 690, 671, 650, 632, 610, 589, 568, 550 da 532 nm ., Sai kawai madaidaicin rafukan da aka raba na rafukan tara a cikin tsari na C9, C8, C7, C6, C5, C4, C3, C2, da C1 suna nunawa.Ana maye gurbin tafki na acrylic da tafkin ma'adini, kuma ana iya lura da ɓangarorin kowane kwararar shunted daga madaidaicin gangara zuwa sama.Bugu da kari, an gyara sub-bidiyo 3 ta yadda za a sake kunna juzu'in canjin raƙuman ruwa na ƙaramin bidiyo 2.Wannan shine mafi kyawun magana na sifofin decochromatic tsararrun madubai.
Sakamakon da ke sama ya nuna cewa tsararrun madubi na decachromatic da aka ƙera ko sabon sikirin mai launi tara yana aiki kamar yadda aka yi niyya.An samar da sabon nau'in sifofi mai launi tara ta hanyar hawa ɗimbin madubai na decachromatic tare da adaftan kai tsaye a kan allon firikwensin hoton.
Haske mai haske tare da kewayon tsayin tsayi daga 400 zuwa 750 nm, wanda aka fitar da maki huɗu na φ50 μm, wanda ke cikin tazara na 1 mm a cikin madaidaiciyar jirgin sama na siffa 2c, bi da bi Binciken 31, 34. Tsarin ruwan tabarau huɗu ya ƙunshi tsararrun ruwan tabarau. ruwan tabarau hudu φ1 mm tare da tsayin daka na 1.4 mm da farar 1 mm.Rafukan da aka haɗu guda huɗu (C0 huɗu) sun faru akan DP na sabon nau'in sifofi mai launi tara, wanda aka raba a tazarar 1 mm.Tsare-tsare na madubin dichroic yana raba kowane rafi (C0) zuwa kogunan launi guda tara (C1-C9).Sakamakon rafukan 36 (tsari huɗu na C1-C9) ana allura kai tsaye zuwa cikin firikwensin hoto na CMOS (S) kai tsaye da aka haɗa kai tsaye zuwa tsararrun madubin dichroic.A sakamakon haka, kamar yadda aka nuna a cikin Fig. 5a, saboda ƙananan matsakaicin matsakaicin matsakaicin hanya mai mahimmanci da kuma gajeren hanya mai mahimmanci, an gano hotunan dukkanin rafukan 36 a lokaci guda kuma a fili tare da girman girman.Dangane da sifofi na ƙasa (duba Ƙarin Hoto S4), ƙarfin hoton rukunoni huɗu C1, C2 da C3 yana da ɗan ƙaranci.Hotuna talatin da shida sun kasance 0.57 ± 0.05 mm cikin girman (ma'anar ± SD).Don haka, girman girman hoton ya kai 11.4.Matsakaicin tazara tsakanin hotuna matsakaita 1 mm (tazara ɗaya kamar jeri na ruwan tabarau) da matsakaicin tazara a kwance 1.6 mm (tazara ɗaya da tsararrun madubi na dichroic).Saboda girman hoton ya fi nisa tsakanin hotuna da yawa, kowane hoto ana iya auna shi da kansa (tare da ƙaramin magana).A halin yanzu, an nuna hotunan rafuka ashirin da takwas da na'urar sikirin launi bakwai na al'ada da aka yi amfani da ita a cikin bincikenmu na baya ana nuna su a cikin siffa 5 B. An ƙirƙiri tsararrun madubai guda bakwai ta hanyar cire madubai biyu mafi kyau na dichroic daga jeri na dichroic tara. madubi a hoto na 1a.Ba duk hotuna ne masu kaifi ba, girman hoton yana ƙaruwa daga C1 zuwa C7.Hotuna ashirin da takwas sune 0.70 ± 0.19 mm a girman.Saboda haka, yana da wuya a kula da babban ƙudurin hoto a cikin duk hotuna.Matsakaicin bambancin (CV) don girman hoto 28 a cikin Hoto 5b shine 28%, yayin da CV don girman hoto 36 a cikin Hoto 5a ya ragu zuwa 9%.Sakamakon da ke sama ya nuna cewa sabon spectrometer mai launi tara ba kawai yana ƙara adadin launuka masu auna lokaci guda daga bakwai zuwa tara ba, amma kuma yana da babban ƙudurin hoto ga kowane launi.
Kwatanta ingancin rarrabuwar hoton da aka kirkira ta al'ada da sabbin spectrometers.(a) Ƙungiyoyi huɗu na hotuna masu raba launi tara (C1-C9) waɗanda sabon nau'in sifofi mai launi tara ya haifar.(b) Rubuce-rubuce huɗu na hotuna masu raba launi bakwai (C1-C7) waɗanda aka kafa tare da na'urar gani mai launi bakwai na al'ada.Fluxes (C0) tare da tsayin raƙuman ruwa daga 400 zuwa 750 nm daga wuraren fitarwa huɗu an haɗu kuma suna faruwa akan kowane sitiriyo, bi da bi.
An yi la'akari da halaye na nau'i na nau'i na nau'i na nau'in launi guda tara da gwaji kuma an nuna sakamakon kimantawa a cikin Hoto 6. Yi la'akari da cewa Hoto 6a yana nuna sakamako iri ɗaya kamar Hoto 5a, watau a tsawon 4 C0 400-750 nm, duk hotunan 36 an gano su. (Kungiyoyi 4 C1-C9).Akasin haka, kamar yadda aka nuna a cikin siffa 6b-j, lokacin da kowane C0 yana da takamaiman tsayin 530, 550, 570, 590, 610, 630, 650, 670, ko 690 nm, akwai kusan hotuna huɗu masu dacewa (hudu). Ƙungiyoyin da aka gano C1, C2, C3, C4, C5, C6, C7, C8 ko C9).Duk da haka, wasu daga cikin hotunan da ke kusa da hotuna guda hudu masu kama da juna suna da rauni sosai saboda yanayin watsawar C1-C9 da aka nuna a cikin Fig. 4b ya dan kadan kuma kowane C0 yana da nau'i na 10 nm a wani takamaiman tsayin daka kamar yadda aka bayyana a cikin hanyar.Waɗannan sakamakon sun yi daidai da sifofin watsa C1-C9 da aka nuna a cikin Figs.4b da ƙarin bidiyo 2 da 3. A wasu kalmomi, na'ura mai launi na tara yana aiki kamar yadda aka sa ran bisa sakamakon da aka nuna a fig.4b ku.Sabili da haka, an kammala cewa rarraba girman hoton C1-C9 shine bakan kowane C0.
Halayen bakan na sikirin mai launi tara.Sabuwar spectrometer mai launi tara yana haifar da saiti huɗu na hotuna masu raba launi tara (C1-C9) lokacin da hasken abin ya faru (C0 huɗu) yana da tsawon (a) 400-750 nm (kamar yadda aka nuna a Hoto 5a), (b) 530nm ku.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, bi da bi.
An yi amfani da na'urar sikeli mai launi tara don electrophoresis mai capillary hudu (don cikakkun bayanai, duba Ƙarin Materials)31,34,35.Matrix na capillary hudu ya ƙunshi capillaries guda huɗu (diamita na waje 360 μm da diamita na ciki 50 μm) waɗanda ke cikin tazara na 1 mm a wurin watsa iska ta Laser.Samfuran da ke ɗauke da gutsuttssun DNA da aka yi wa lakabi da rini guda 8, wato FL-6C (dye 1), JOE-6C (dye 2), dR6G (dye 3), TMR-6C ( rini 4), CXR-6C ( rini 5), TOM- 6C (dye 6), LIZ (dye 7), da WEN (dye 8) a cikin tsarin hawan hawan igiyar ruwa, sun rabu a cikin kowane capillaries hudu (wanda ake kira Cap1, Cap2, Cap3, da Cap4).Hasken haske mai haifar da Laser daga Cap1-Cap4 an haɗa shi tare da tsararrun ruwan tabarau guda huɗu kuma an yi rikodin lokaci guda tare da na'urar gani mai launi tara.Matsakaicin ƙarfin ƙarfin haske mai launi tara (C1-C9) a lokacin electrophoresis, wato, electrophoregram mai launi tara na kowane capillary, an nuna shi a cikin siffa 7a.Ana samun daidaitaccen electrophoregram mai launi tara a cikin Cap1-Cap4.Kamar yadda kibiyoyin Cap1 suka nuna a cikin Hoto 7a, kololuwa takwas akan kowane electrophoregram mai launi tara suna nuna fitar da kyalli ɗaya daga Dye1-Dye8, bi da bi.
Ƙididdigar lokaci ɗaya na rinayen guda takwas ta amfani da na'urar sikeli mai launi huɗu mai launi tara.(a) Launuka tara (C1-C9) electrophoregram na kowane capillary.Kololuwa takwas da kibiyoyi Cap1 ke nunawa suna nuna fitar da hayaki mai haske na rini takwas (Dye1-Dye8).Launukan kiban sun dace da launuka (b) da (c).(b) Bakan haske na rini takwas (Dye1-Dye8) a kowace capillary.c Electropherograms na rini takwas (Dye1-Dye8) kowace capillary.Kololuwar ɓangarorin DNA masu alamar Dye7 ana nuna su ta kibiyoyi, kuma ana nuna tsayin tushe na Cap4.
An nuna ƙarfin rabon C1-C9 akan kololuwa takwas a cikin Fig.7b, ku.Saboda duka C1-C9 da Dye1-Dye8 suna cikin tsari mai tsayi, rarrabawar takwas a cikin siffa 7b suna nuna bakan haske na Dye1-Dye8 a jere daga hagu zuwa dama.A cikin wannan binciken, Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, da Dye8 sun bayyana a cikin magenta, violet, blue, cyan, green, yellow, orange, da ja, bi da bi.Lura cewa launukan kiban da ke cikin siffa 7a sun dace da launukan rini a hoto na 7b.Ƙarfin hasken wuta na C1-C9 na kowane bakan a cikin Hoto 7b an daidaita su ta yadda jimlar su ta zama ɗaya.An samo nau'ikan nau'ikan kyalli takwas daidai daga Cap1-Cap4.Mutum na iya lura da abin da ya faru a fili tsakanin rini 1- rini 8.
Kamar yadda aka nuna a cikin Hoto 7c, ga kowane capillary, electrophoregram mai launi tara a cikin Hoto 7a an canza shi zuwa na'urar lantarki mai launi takwas ta hanyar bincike mai yawa dangane da bakan haske takwas a cikin Hoto 7b (duba Ƙarin Materials don cikakkun bayanai).Tunda ba'a nuna bayyani a cikin Hoto na 7a a cikin Hoto na 7a, Dye1-Dye8 za'a iya ganowa da ƙididdige su daban-daban a kowane lokaci, ko da mabambantan nau'ikan fluoresce na Dye1-Dye8 a lokaci guda.Ba za a iya yin wannan tare da gano launi bakwai na gargajiya31 ba, amma ana iya samun su tare da haɓakar gano launuka tara.Kamar yadda aka nuna ta kiban Cap1 a cikin siffa 7c, kawai masu fitar da iska mai kyalli Dye3 (blue), Dye8 (ja), Dye5 (kore), Dye4 (cyan), Dye2 (purple), Dye1 (magenta), da Dye6 (Yellow). ) ana lura da su a cikin tsarin da ake tsammani.Don fitowar mai kyalli na rini 7 (orange), baya ga kololuwa guda ɗaya da kibiyar lemu ta nuna, an ga wasu kololuwa da yawa.Wannan sakamakon ya faru ne saboda gaskiyar cewa samfurori sun ƙunshi ma'auni masu girma, Dye7 mai lakabin DNA tare da tsayin tushe daban-daban.Kamar yadda aka nuna a cikin Hoto 7c, don Cap4 waɗannan tsayin tushe sune 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 da 220 tushe.
Babban fasali na spectrometer mai launi tara, wanda aka haɓaka ta amfani da matrix na madubai na dichroic dichroic Layer biyu, ƙananan girman da zane mai sauƙi.Tun da tsararrun madubin decachromatic a cikin adaftar da aka nuna a fig.3c wanda aka ɗora kai tsaye a kan allon firikwensin hoto (duba siffa S1 da S2), ma'aunin launi guda tara yana da ma'auni iri ɗaya da adaftar, watau 54 × 58 × 8.5 mm.(kauri) .Wannan matsananci-kananan girman umarni biyu zuwa uku ne na girman girma fiye da na'urori masu aunawa na al'ada waɗanda ke amfani da gratings ko prisms.Bugu da ƙari, tun da an saita na'urar sikelin launi tara ta yadda haske ya faɗo saman firikwensin hoto a kai a kai, ana iya keɓance sarari cikin sauƙi don ma'aunin sifofi mai launi tara a cikin tsarin kamar microscopes, cytometers masu gudana, ko masu nazari.Capillary grating electrophoresis analyzer don ko da mafi girma miniaturization na tsarin.A lokaci guda, girman madubin dichroic guda goma da matattarar bandpass da aka yi amfani da su a cikin spectrometer mai launi tara kawai 10 × 1.9 × 0.5 mm ko 15 × 1.9 × 0.5 mm.Don haka, fiye da 100 irin waɗannan ƙananan madubai na dichroic da matattarar bandpass, bi da bi, ana iya yanke su daga madubi dichroic da matatar bandpass 60 mm2, bi da bi.Don haka, ana iya kera jigon madubin decachromatic akan farashi mai rahusa.
Wani fasali na ma'aunin siffa mai launi tara shine kyawawan halayensa na gani.Musamman ma, yana ba da damar siyan hotuna masu kyan gani na hotuna, wato, sayan hotuna a lokaci guda tare da bayanan kallo.Ga kowane hoto, an sami ci gaba da bakan tare da kewayon tsayi daga 520 zuwa 700 nm da ƙuduri na 20 nm.A wasu kalmomi, ana gano ƙarfin haske masu launi tara ga kowane hoto, watau maƙallan 20nm guda tara daidai da raba nisa daga 520 zuwa 700 nm.Ta hanyar canza halayen madubi na dichroic da matatar bandpass, za a iya daidaita kewayon tsayin igiyoyi tara da faɗin kowace ƙungiya.Ana iya amfani da gano launi tara ba kawai don ma'aunin haske ba tare da hoto na gani (kamar yadda aka bayyana a cikin wannan rahoton), amma har ma da sauran aikace-aikacen gama gari da yawa ta amfani da hoto mai ban mamaki.Kodayake hotunan hyperspectral na iya gano ɗaruruwan launuka, an gano cewa ko da tare da raguwa mai yawa a cikin adadin launuka masu iya ganewa, ana iya gano abubuwa da yawa a cikin filin ra'ayi tare da isasshen daidaito don yawancin aikace-aikace38,39,40.Saboda ƙudurin sararin samaniya, ƙudirin gani, da ƙudirin ɗan lokaci suna da ciniki a cikin hoto, rage yawan launuka na iya haɓaka ƙudurin sararin samaniya da ƙudurin ɗan lokaci.Hakanan yana iya amfani da na'urori masu sauƙi kamar wanda aka haɓaka a cikin wannan binciken kuma yana ƙara rage yawan ƙididdiga.
A cikin wannan binciken, an ƙididdige rinannun guda takwas a lokaci guda ta hanyar rarrabuwar kawuna na nau'ikan kyamarorinsu da suka mamaye bisa ga gano launuka tara.Ana iya ƙididdige rini har zuwa tara a lokaci guda, suna zama tare cikin lokaci da sarari.Fa'ida ta musamman na sikirin mai launi tara shine babban jujjuyawar haske da babban buɗewar (1 × 7 mm).Tsare-tsare na madubi mai lalacewa yana da matsakaicin watsawa na 92% na haske daga buɗaɗɗen kewayo a cikin kowane jeri tara.Ingancin amfani da hasken abin da ya faru a cikin kewayon tsayi daga 520 zuwa 700 nm kusan 100%.A cikin irin wannan fadi da kewayon raƙuman raƙuman raƙuman ruwa, babu diffraction grating da zai iya samar da irin wannan babban ingancin amfani.Ko da ingancin diffraction grating na diffraction ya wuce 90% a wani ɗan gajeren zango, yayin da bambanci tsakanin wannan tsayin da wani tsayin tsayin tsayin raƙuman raƙuman ruwa yana raguwa41.Faɗin buɗaɗɗen daidai gwargwado ga alkiblar jirgin a cikin siffa 2c na iya tsawanta daga 7 mm zuwa faɗin firikwensin hoto, kamar a yanayin firikwensin hoton da aka yi amfani da shi a cikin wannan binciken, ta ɗan gyaggyara tsararrun decamer.
Za a iya amfani da na'urar spectrometer mai launi tara ba kawai don capillary electrophoresis ba, kamar yadda aka nuna a cikin wannan binciken, har ma da wasu dalilai daban-daban.Misali, kamar yadda aka nuna a cikin hoton da ke ƙasa, ana iya amfani da na'urar sikirin mai launi tara akan ma'aunin haske.Ana nuna jirgin samfurin akan firikwensin hoton na sikirin mai launi tara ta hanyar manufa 10x.Nisa na gani tsakanin madaidaicin ruwan tabarau da firikwensin hoto shine mm 200, yayin da nisa na gani tsakanin abin da ya faru na spectrometer mai launi tara da firikwensin hoto shine mm 12 kawai.Sabili da haka, an yanke hoton zuwa kimanin girman girman budewa (1 × 7 mm) a cikin jirgin da ya faru kuma an raba shi zuwa hotuna masu launi tara.Wato, ana iya ɗaukar hoto mai ban mamaki na hoto mai launi tara akan yanki 0.1 × 0.7 mm a cikin samfurin jirgin sama.Bugu da ƙari, yana yiwuwa a sami hoto mai launi mai launi tara na yanki mafi girma a kan samfurin jirgin sama ta hanyar nazarin samfurin dangane da maƙasudin a cikin layi na kwance a cikin Fig. 2c.
Abubuwan tsararrun madubi na decachromatic, wato M1-M9 da BP, Asahi Spectra Co., Ltd. ne ya yi su ta hanyar amfani da daidaitattun hanyoyin hazo.Multilayer dielectric kayan an yi amfani da akayi daban-daban a kan goma ma'adini faranti 60 × 60 mm a size da kuma 0.5 mm kauri, saduwa da wadannan bukatun: M1: IA = 45 °, R ≥ 90% a 520-590 nm, Tave ≥ 90% a 610- 610nm ku.700 nm, M2: IA = 45 °, R ≥ 90% a 520-530 nm, Tave ≥ 90% a 550-600 nm, M3: IA = 45 °, R ≥ 90% a 540-550 nm, Tave 90≥ % a 570-600 nm, M4: IA = 45 °, R ≥ 90% a 560-570 nm, Tave ≥ 90% a 590-600 nm, M5: IA = 45 °, R ≥ 98% a 580-600 nm , R ≥ 98% a 680-700 nm, M6: IA = 45 °, Tave ≥ 90% a 600-610 nm, R ≥ 90% a 630-700 nm, M7: IA = 45 °, R ≥ a 90% 620-630 nm, Taw ≥ 90% a 650-700 nm, M8: IA = 45 °, R ≥ 90% a 640-650 nm, Taw ≥ 90% a 670-700 nm, M9: IA = 45 ° ≥ 90% a 650-670 nm, Tave ≥ 90% a 690-700 nm, BP: IA = 0 °, T ≤ 0.01% a 505 nm, Tave ≥ 95% a 530-690 nm a 530 nm a -690 nm da T ≤ 1% a 725-750 nm, inda IA, T, Tave, da R sune kusurwar abin da ya faru, watsawa, matsakaicin watsawa, da haske mai haske.
Farin haske (C0) tare da kewayon tsayin 400-750 nm wanda tushen hasken LED (AS 3000, AS ONE CORPORATION) ya haɗu kuma ya faru a tsaye akan DP na tsararrun madubin dichroic.Ana nuna bakan haske na farin LEDs a Ƙarin Hoto S3.Sanya tanki na acrylic (girman 150 × 150 × 30 mm) kai tsaye a gaban tsararrun madubi na decamera, gaban PSU.Hayakin da aka samu lokacin da busasshen ƙanƙara ya nutsar da shi a cikin ruwa an zuba shi a cikin tankin acrylic don lura da raƙuman raƙuman ruwa na C1-C9 masu launi tara waɗanda ke fitowa daga tsararrun madubai na decachromatic.
A madadin, an haɗa farin haske (C0) ta hanyar tacewa kafin shigar da DP.Tace masu tacewa asalin matatun mai yawa ne na tsaka tsaki tare da ƙimar gani na 0.6.Sannan yi amfani da matatar mai motsi (FW212C, FW212C, Thorlabs).A ƙarshe, kunna tace ta ND baya.Matsakaicin bandwidth na matatun bandpass tara sun dace da C9, C8, C7, C6, C5, C4, C3, C2 da C1, bi da bi.An sanya tantanin ma'adini mai girma na ciki na 40 (tsawon gani) x 42.5 (tsawo) x 10 mm (nisa) a gaban tsararrun madubai na decochromatic, sabanin BP.Ana ciyar da hayakin ta cikin bututu a cikin tantanin halitta na ma'adini don kiyaye yawan hayaki a cikin tantanin halitta na quartz don ganin rafukan raba C1-C9 masu launi tara waɗanda ke fitowa daga tsararrun madubi na decachromatic.
Bidiyo na rafin haske mai rabe-raben launuka tara da ke fitowa daga ɗimbin madubai na lalata an kama shi cikin yanayin rashin lokaci akan iPhone XS.Ɗauki hotuna na wurin a 1fps kuma tattara hotuna don ƙirƙirar bidiyo a 30fps (don bidiyo na zaɓi 1) ko 24fps (don bidiyo na zaɓi 2 da 3).
Sanya farantin bakin karfe mai kauri 50µm (tare da ramukan diamita 50 µm guda hudu a tazara 1 mm) akan farantin watsawa.Haske mai tsayin tsayin 400-750nm yana haskakawa akan farantin mai watsawa, wanda aka samu ta hanyar wucewar haske daga fitilar halogen ta hanyar gajeriyar watsawa tare da yanke kauri na 700 nm.Ana nuna bakan haske a Ƙarin Hoto S4.A madadin, hasken kuma yana wucewa ta ɗaya daga cikin 10 nm matatar bandpass wanda ke tsakiya a 530, 550, 570, 590, 610, 630, 650, 670 da 690 nm kuma ya buga farantin mai watsawa.Sakamakon haka, an samar da maki huɗu masu radiyo tare da diamita na φ50 μm da tsayi daban-daban akan farantin bakin karfe da ke gaban farantin mai yaduwa.
An ɗora tsararru na capillary guda huɗu tare da ruwan tabarau guda huɗu akan nau'in sifofi mai launi tara kamar yadda aka nuna a Figures 1 da 2. C1 da C2.Rubutun capillaries guda huɗu da ruwan tabarau huɗu sun kasance daidai da na karatun baya31,34.Laser katako mai tsayi na 505 nm da ƙarfin 15 mW yana haskakawa lokaci guda kuma a ko'ina daga gefe zuwa wuraren fitar da capillaries hudu.Hasken walƙiya da ke fitarwa ta kowane wurin fitarwa yana haɗuwa da ruwan tabarau mai dacewa kuma an raba shi zuwa kogunan launi guda tara ta hanyar madubi na decachromatic.Sakamakon rafukan 36 da aka samu sannan an yi musu allurar kai tsaye cikin firikwensin hoton CMOS (C11440-52U, Hamamatsu Photonics K·K.), kuma an yi rikodin hotunansu lokaci guda.
ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Aikace-aikacen Halittun Halitta), 4 µl GeneScan™ 600 LIZ™ an gauraye rini ga kowane capillary ta hanyar hada 1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), 1 µl ma'aunin girman girman.v2.0 (Thermo Fisher Scientific) da 14µl na ruwa.Ma'aunin Matrix na PowerPlex® 6C ya ƙunshi guntu DNA guda shida waɗanda aka yi wa lakabi da rinai shida: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, da WEN, a cikin tsari mafi girman tsayi.Ba a bayyana tsawon tushe na waɗannan guntuwar DNA ba, amma an san jerin tsawon tushe na guntun DNA da aka yi wa lakabi da WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C da TOM-6C.Haɗin da ke cikin ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit yana ƙunshe da guntun DNA da aka yiwa lakabi da dR6G rini.Hakanan ba a bayyana tsayin tushe na guntuwar DNA ba.GeneScan™ 600 LIZ™ Dye Size Standard v2.0 ya haɗa da gutsuttsuran DNA masu alamar LIZ guda 36.Tsawon tushe na waɗannan gutsutsayen DNA sune 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 30,30, 30,300 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 da 600 tushe.An cire samfuran a 94 ° C na mintuna 3, sannan a sanyaya su akan kankara na mintuna 5.An allura samfurori a cikin kowane capillary a 26 V / cm na 9 s kuma an raba su a cikin kowane capillary da aka cika da POP-7 ™ polymer bayani (Thermo Fisher Scientific) tare da tsayin tasiri na 36 cm da ƙarfin lantarki na 181 V / cm da kwana na 60°.DAGA
Duk bayanan da aka samu ko aka yi nazari a cikin wannan binciken suna cikin wannan labarin da aka buga da ƙarin bayanansa.Ana samun wasu bayanan da suka dace da wannan binciken daga marubutan bisa ga buƙatu mai ma'ana.
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