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Hōʻike i kahi carousel o ʻekolu paheʻe i ka manawa hoʻokahi.E hoʻohana i nā pihi Mua a me Next no ka neʻe ʻana i ʻekolu paheʻe i ka manawa, a i ʻole e hoʻohana i nā pihi paheʻe ma ka hopena e neʻe i ʻekolu mau paheʻe i ka manawa.
Ua hoʻomohala ʻia kahi ʻano kikoʻī ultra-compact (54 × 58 × 8.5 mm) a me ka puka ākea (1 × 7 mm) ʻeiwa ʻulaʻula, "māhele ʻia i ʻelua" e kahi hui o nā aniani dichroic he ʻumi, i hoʻohana ʻia no ke kiʻi kikoʻī koke.Hoʻokaʻawale ʻia ka ʻāpana māmā me ka ʻāpana liʻiliʻi ma mua o ka nui o ka aperture i kahi ʻāpana hoʻomau 20 nm ākea a ʻeiwa mau kala kala me nā nalu waena o 530, 550, 570, 590, 610, 630, 650, 670 a me 690 nm.Hoʻohana maikaʻi ʻia nā kiʻi o nā kahawai ʻeiwa i ka manawa like e ka mea ʻike kiʻi.ʻAʻole like me nā ʻāpana aniani dichroic maʻamau, ʻo ka hoʻolālā aniani dichroic i kūkulu ʻia he ʻano hoʻonohonoho ʻelua ʻāpana, ʻaʻole ia e hoʻonui wale i ka helu o nā kala i hiki ke ana ʻia i ka manawa like, akā hoʻomaikaʻi pū i ka hoʻonā kiʻi no kēlā me kēia kahawai kala.Hoʻohana ʻia ka spectrometer ʻeiwa kala i kūkulu ʻia no ka electrophoresis ʻehā-capillary.ʻO ka hoʻopaʻa ʻana helu helu like ʻana o nā mea pena ʻewalu e neʻe like ana i kēlā me kēia capillary me ka hoʻohana ʻana i ka fluorescence laser-induced ʻeiwa kala.No ka mea, ʻaʻole liʻiliʻi a maʻalahi ka spectrometer ʻeiwa kala, akā he kiʻekiʻe luminous flux a lawa ka spectral hoʻonā no ka hapa nui o nā noi kiʻi spectral, hiki ke hoʻohana nui ʻia i nā ʻano āpau.
Ua lilo ke kiʻi hyperspectral a me multispectral i mea nui o ka astronomy2, ka ʻike mamao no ka nānā ʻana i ka Honua3,4, ka mālama ʻana i ka maikaʻi o ka meaʻai a me ka wai5,6, mālama kiʻi a me ka archeology7, forensics8, surgery9, biomedical analysis and diagnostics10,11 etc. Field 1 He ʻenehana pono ʻole. ,12,13.Hoʻokaʻawale ʻia nā ʻano hana no ke ana ʻana i ka spectrum o ka mālamalama i hoʻokuʻu ʻia e kēlā me kēia wahi o ka hoʻokuʻu ʻana i ke kahua o ka nānā ʻana i (1) mākaʻikaʻi kiko ("broom")14,15, (2) scanning linear ("panicle")16,17,18 , (3) nānā ka lōʻihi i nā nalu19,20,21 a me (4) kiʻi22,23,24,25.I ka hihia o kēia mau ʻano a pau, ʻo ka hoʻonā spatial, spectral resolution a me ke kino hoʻonā i loaʻa kahi pilina kālepa9,10,12,26.Eia kekahi, he hopena koʻikoʻi ka puka māmā i ka naʻau, ʻo ia hoʻi ka ratio signal-to-noise i ka spectral imaging26.ʻO ka luminous flux, ʻo ia hoʻi, ka maikaʻi o ka hoʻohana ʻana i ka mālamalama, pili pono i ka ratio o ka nui i ana ʻia o ka mālamalama o kēlā me kēia kiko kukui i kēlā me kēia manawa i ka nui o ka mālamalama o ka laulā hawewe i ana ʻia.ʻO ka Māhele (4) he ala kūpono ke hoʻololi ʻia ka ikaika a i ʻole ke kikoo o ka mālamalama i hoʻopuka ʻia e kēlā me kēia kiko hoʻokuʻu me ka manawa a i ʻole ke hoʻololi ʻia ke kūlana o kēlā me kēia kiko hoʻokuʻu me ka manawa no ka mea ua ana ʻia ke ʻano o ke kukui i hoʻopuka ʻia e nā wahi hoʻokuʻu a pau.24.
Hoʻohui ʻia ka hapa nui o nā ʻano i luna me nā spectrometers nui, paʻakikī a / a pipiʻi paha me ka hoʻohana ʻana i 18 gratings a i ʻole 14, 16, 22, 23 prisms no nā papa (1), (2) a me (4) a i ʻole 20, 21 kānana kānana, nā kānana wai. .Nā kānana hiki ke hoʻoheheʻe ʻia (LCTF)25 a i ʻole nā kānana hoʻokani acousto-optic (AOTF)19 o ka māhele (3).ʻO ka ʻokoʻa, ʻano liʻiliʻi a ʻuʻuku nā spectrometers multi-mirror ma muli o kā lākou hoʻonohonoho maʻalahi27,28,29,30.Eia kekahi, loaʻa iā lākou kahi ʻōlinolino kiʻekiʻe no ka mea ua hoʻohana piha ʻia ka mālamalama i hoʻokaʻawale ʻia e kēlā me kēia aniani dichroic (ʻo ia hoʻi, ke kukui i hoʻouna ʻia a hōʻike ʻia o ke kukui hanana ma kēlā me kēia aniani dichroic).Eia nō naʻe, ʻo ka helu o nā pūnaʻi hawewe (ʻo ia hoʻi nā kala) pono e ana ʻia i ka manawa like, ua kaupalena ʻia i ʻehā paha.
Hoʻohana mau ʻia ke kiʻi spectral e pili ana i ka ʻike fluorescence no ka loiloi multiplex i ka ʻike biomedical a me nā diagnostics 10, 13.I ka multiplexing, no ka mea, ua kapa 'ia na analytes he nui (e la'a, DNA a i 'ole nā proteins) me nā mea ho'onani fluorescent like 'ole, ua helu 'ia kēlā me kēia analyte i kēlā me kēia wahi ho'oku'u ma ke kahua o ka nānā 'ana me ka ho'ohana 'ana i ka multicomponent analysis.32 wawahi iho i ke kikoo fluorescence i ikeia i hoopukaia e kela me keia wahi hookui.I loko o kēia kaʻina hana, hiki i nā kala like ʻole, kēlā me kēia mea e hoʻopuka ana i kahi fluorescence ʻokoʻa, hiki ke colocalize, ʻo ia hoʻi, noho pū i ka lewa a me ka manawa.I kēia manawa, ʻo ka helu kiʻekiʻe o nā mea pena e hiki ke hoʻohauʻoli ʻia e ka kukuna laser hoʻokahi he 833.ʻAʻole i hoʻoholo ʻia kēia palena kiʻekiʻe e ka hoʻonā spectral (ʻo ia hoʻi, ka helu o nā kala), akā ma ka laulā o ka spectrum fluorescence (≥50 nm) a me ka nui o ka dye Stokes shift (≤200 nm) ma FRET (hoʻohana i ka FRET)10 .Eia naʻe, pono e ʻoi aku ka nui o nā waihoʻoluʻu ma mua a i ʻole like me ka helu o nā mea pena e hoʻopau ai i ka uhi ʻana o nā kala i hui ʻia31,32.No laila, pono e hoʻonui i ka helu o nā kala i ana ʻia i ka ʻewalu a ʻoi aku paha.
I kēia mau lā, ua hoʻomohala ʻia kahi spectrometer heptachroic ultra-compact (e hoʻohana ana i kahi ʻano o nā aniani heptychroic a me kahi mea ʻike kiʻi e ana i ʻehā mau fluorescent fluxes).ʻO ka spectrometer ʻelua a ʻekolu mau kauoha o ka nui ka liʻiliʻi ma mua o nā spectrometer maʻamau e hoʻohana ana i nā gratings a i ʻole prisms34,35.Eia naʻe, paʻakikī ke kau ʻana ma mua o ʻehiku mau aniani dichroic i loko o kahi spectrometer a i ka manawa like e ana ʻoi aku ma mua o ʻehiku kala36,37.Me ka hoʻonui ʻana i ka helu o nā aniani dichroic, piʻi ka ʻokoʻa kiʻekiʻe o ka lōʻihi o nā ala ʻike o nā kukui kukui dichroic, a lilo ia i mea paʻakikī ke hōʻike i nā kahe kukui āpau ma kahi mokulele sensory.Piʻi aʻe ka lōʻihi o ke ala ʻoi loa o ka māmā, no laila e hoʻemi ana ka laulā o ka puka spectrometer (ʻo ia hoʻi ka laulā kiʻekiʻe o ke kukui i kālai ʻia e ka spectrometer).
I ka pane ʻana i nā pilikia i luna aʻe, ua kūkulu ʻia kahi spectrometer ʻeiwa ʻulaʻula ultra-compact me kahi ʻāpana aniani decachromatic "dichroic" ʻelua ʻāpana a me kahi mea ʻike kiʻi no ke kiʻi kikoʻī koke [kāhua (4)].Hoʻohālikelike ʻia me nā spectrometers mua, ʻoi aku ka liʻiliʻi o ka spectrometer i hoʻomohala ʻia i ka lōʻihi o ke ala optical kiʻekiʻe a ʻoi aku ka liʻiliʻi o ka lōʻihi ala optical.Ua hoʻohana ʻia i nā electrophoresis ʻehā-capillary e ʻike i ka fluorescence ʻeiwa kala i hoʻokomo ʻia i ka laser a me ka helu ʻana i ka neʻe like ʻana o ʻewalu mau kala i kēlā me kēia capillary.No ka mea ʻaʻole liʻiliʻi a maʻalahi ka spectrometer i kūkulu ʻia, akā loaʻa nō hoʻi kahi flux luminous kiʻekiʻe a lawa ka hoʻonā spectral no ka hapa nui o nā noi kiʻi spectral, hiki ke hoʻohana nui ʻia i nā ʻano āpau.
Hōʻike ʻia ka spectrometer ʻeiwa kala maʻamau i ka fig.1a.ʻO kona hoʻolālā e hahai ana i ko ka ultra-liʻiliʻi ʻehiku kala spectrometer 31. Aia i ʻeiwa mau aniani dichroic i hoʻonohonoho ʻia ma ka ʻaoʻao o 45 ° i ka ʻākau, a aia ka ʻike kiʻi (S) ma luna o nā aniani dichroic ʻeiwa.Ua māhele ʻia ke kukui e komo ana mai lalo mai (C0) e kahi ʻeiwa aniani dichroic i ʻeiwa mau kahe kukui e piʻi ana (C1, C2, C3, C4, C5, C6, C7, C8 a me C9).Hānai ʻia nā kahawai ʻeiwa a pau i ka ʻike kiʻi a ʻike ʻia i ka manawa like.Ma kēia haʻawina, ʻo C1, C2, C3, C4, C5, C6, C7, C8, a me C9 ma ke ʻano o ka lōʻihi nalu a ua hōʻike ʻia e ka magenta, violet, blue, cyan, ʻōmaʻomaʻo, melemele, ʻalani, ʻulaʻula-ʻalani, a ʻulaʻula, kēlā me kēia.ʻOiai ua hoʻohana ʻia kēia mau kala kala i kēia palapala, e like me ka hōʻike ʻana ma ke Kiʻi 3, no ka mea, ʻokoʻa lākou mai nā kala maoli i ʻike ʻia e ka maka kanaka.
Nā kiʻi kiʻi kiʻi o nā spectrometers ʻeiwa kala hou.(a) ʻEiwa waihoʻoluʻu spectrometer maʻamau me ʻeiwa mau aniani dichroic.(b) ʻEiwa kala hou me kahi ʻāpana aniani dichroic ʻelua.Ua māhele ʻia ka hanana kukui C0 i ʻeiwa mau kala kukui C1-C9 a ʻike ʻia e ka mea ʻike kiʻi S.
ʻO ka spectrometer ʻeiwa ʻulaʻula hou i kūkulu ʻia he ʻelua ʻāpana aniani dichroic grating a me kahi ʻike kiʻi, e like me ka hōʻike ʻana ma ka Fig. 1b.Ma ka papa haʻahaʻa, ʻelima mau aniani dichroic i hili ʻia i ka 45° i ka ʻākau, e hoʻopili ʻia i ka ʻākau mai ke kikowaena o ka hui decamers.Ma ka pae kiʻekiʻe, ʻelima mau aniani dichroic hou e hili ʻia 45° i ka hema a aia mai ke kikowaena a i ka hema.ʻO ke aniani dichroic hema loa o ka papa haʻahaʻa a me ke aniani dichroic ʻākau loa o ka ʻaoʻao luna e uhi kekahi i kekahi.Ua māhele ʻia ka ʻāwili kukui hanana (C0) mai lalo mai i ʻehā mau aniani chromatic puka waho (C1-C4) e nā aniani dichroic ʻelima ma ka ʻākau a me ʻelima mau aniani chromatic puka waho (C5-C4) e nā aniani dichroic ʻelima ma ka hema C9).E like me nā spectrometers ʻeiwa kala maʻamau, hoʻokomo pololei ʻia nā kahawai ʻeiwa a pau i loko o ka mea ʻike kiʻi (S) a ʻike ʻia i ka manawa like.I ka hoʻohālikelike ʻana i nā Kiʻi 1a a me 1b, hiki i kekahi ke ʻike i ke ʻano o ka spectrometer ʻeiwa ʻulaʻula hou, ʻo ka ʻokoʻa kiʻekiʻe a me ka lōʻihi o ka lōʻihi o ke ala ʻoi loa o nā ʻeiwa waihoʻoluʻu e hapalua.
Hōʻike ʻia ke kūkulu kikoʻī ʻana o kahi pūʻulu aniani dichroic ultra-liʻiliʻi ʻelua ʻāpana 29 mm (ākea) × 31 mm (hohonu) × 6 mm (kiʻekiʻe) ma ke Kiʻi 2. ʻO ka hui aniani dichroic decimal he ʻelima mau aniani dichroic ma ka ʻākau. (M1-M5) a me ʻelima mau aniani dichroic ma ka hema (M6-M9 a me kekahi M5), ua hoʻopaʻa ʻia kēlā me kēia aniani dichroic i ka bracket alumini luna.Hoʻopili ʻia nā aniani dichroic āpau e uku no ka neʻe ʻana like ʻana ma muli o ka refraction o ke kahe ma o nā aniani.Ma lalo o M1, ua hoʻopaʻa ʻia kahi kānana band-pass (BP).ʻO ka nui M1 a me BP he 10mm (ʻaoʻao lōʻihi) x 1.9mm (ʻaoʻao pōkole) x 0.5mm (mānoanoa).ʻO nā ana o nā aniani dichroic i koe he 15 mm × 1.9 mm × 0.5 mm.ʻO ka pitch matrix ma waena o M1 a me M2 he 1.7 mm, aʻo ka pitch matrix o nā aniani dichroic ʻē aʻe he 1.6 mm.Ma ka fig.Hoʻohui ʻo 2c i ke kahe kukui C0 a me ʻeiwa mau kukui kala ʻulaʻula C1-C9, i hoʻokaʻawale ʻia e kahi matrix de-chamber o nā aniani.
Ke kūkulu ʻia ʻana o kahi matrix aniani dichroic ʻelua.(a) He hiʻohiʻona hiʻohiʻona a me (b) he ʻike ʻaoʻao ʻaoʻao o kahi ʻāpana aniani dichroic ʻelua ʻāpana (nā ana 29 mm x 31 mm x 6 mm).He ʻelima mau aniani dichroic (M1-M5) aia ma ka papa haʻahaʻa, ʻelima mau aniani dichroic (M6-M9 a me kekahi M5) aia ma ka papa luna, a me kahi kānana bandpass (BP) aia ma lalo o M1.(c) Nānā ʻaoʻao keʻa ma ka ʻaoʻao kū pololei, me C0 a me C1-C9 e kau ana.
ʻO ka laulā o ka puka ma ka ʻaoʻao ākea, i hōʻike ʻia e ka laulā C0 ma ka Fig. 2, c, he 1 mm, a ma ke ala pololei i ka mokulele o Fig. 2, c, i hāʻawi ʻia e ka hoʻolālā ʻana o ka bracket aluminika. – 7 mm.ʻO ia hoʻi, ʻo ka spectrometer ʻeiwa ʻulaʻula hou ka nui aperture nui o 1 mm × 7 mm.ʻO ke ala optical o C4 ka lōʻihi loa ma waena o C1-C9, a ʻo ke ala ʻike o C4 i loko o ke aniani aniani dichroic, ma muli o ka nui liʻiliʻi ma luna (29 mm × 31 mm × 6 mm), he 12 mm.I ka manawa like, ʻo ka lōʻihi o ke ala opua o C5 ka pōkole loa ma waena o C1-C9, a ʻo ka lōʻihi o ke ala optical o C5 he 5.7mm.No laila, ʻo ka ʻokoʻa kiʻekiʻe loa i ka lōʻihi o ke ala optical he 6.3 mm.Hoʻoponopono ʻia nā lōʻihi o ke ala optical no ka lōʻihi o ke ala optical no ka lawe ʻana o M1-M9 a me BP (mai ka quartz).
Ua helu ʻia nā waiwai spectral o М1−М9 a me VR no laila ʻo nā fluxes С1, С2, С3, С4, С5, С6, С7, С8 a me С9 aia i loko o ka laulā nalu 520–540, 540–560, 560–5080, 560–5080 –600 , 600–620, 620–640, 640–660, 660–680, a me 680–700 nm.
Hōʻike ʻia ke kiʻi o ka matrix i hana ʻia o nā aniani decachromatic ma Fig. 3a.Hoʻopili ʻia ʻo M1-M9 a me BP i ka 45-degree slope a me ka mokulele ākea o ke kākoʻo alumini, ʻoiai ua hūnā ʻia ʻo M1 a me BP ma ke kua o ke kiʻi.
Ka hana ʻana o nā aniani decan a me kāna hōʻike.(a) He ʻano o nā aniani decachromatic i hana ʻia.(b) He kiʻi māhele ʻeiwa ʻeiwa he 1 mm × 7 mm i hoʻopaʻa ʻia ma luna o kahi pepa i waiho ʻia i mua o nā aniani decachromatic a me nā kukui keʻokeʻo.(c) He mau aniani decochromatic i hoʻomālamalama ʻia me ke kukui keʻokeʻo mai hope.(d) ʻEiwa ʻōniʻoniʻo māwehe kahawai e puka mai ana mai ke aniani aniani decane, i ʻike ʻia ma ke kau ʻana i ka pahu acrylic piha i ka uahi ma ke alo o ke aniani decane ma ka c a me ka pōʻeleʻele o ka lumi.
Hōʻike ʻia ke ʻano o ka hoʻouna ʻana o M1-M9 C0 ma kahi kihi o ka hanana o 45 ° a me ke ana ʻana o ka hoʻouna ʻana o BP C0 ma kahi kihi o ka hopena o 0 °.4a.Hōʻike ʻia ka spectra hoʻouna o C1-C9 e pili ana iā C0 ma Fig.4b.Ua helu ʻia kēia mau spectra mai ka spectra ma Fig.4a e like me ke ala opua C1-C9 ma Fig. 4a.1b a me 2c.No ka laʻana, 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)], kahi o TS(X) a me [ 1 − TS(X)] ʻo ia ka hoʻouna ʻana a me ka hiʻohiʻona noʻonoʻo o X.E like me ka mea i hōʻike ʻia ma ka Figure 4b, ʻo nā bandwidths (bandwidth ≥50%) o C1, C2, C3, C4, C5, C6, C7, C8 a me C9 he 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 a me 682-699 nm.Kūlike kēia mau hopena me nā pae i kūkulu ʻia.Eia kekahi, ʻoi aku ka kiʻekiʻe o ka hoʻohana ʻana o ke kukui C0, ʻo ia hoʻi, ʻo ka awelika kiʻekiʻe C1-C9 light transmittance he 92%.
Hoʻouna i nā kikoʻī o kahi aniani dichroic a me kahi ʻeiwa kala ʻeiwa.(a) Ana 'ia ka ho'ouna 'ana o M1-M9 ma 45° a me BP ma 0°.(b) Ka hoʻouna kikoʻī o C1–C9 pili i C0 i helu ʻia mai (a).
Ma ka fig.3c, aia ka papa o na aniani aniani dichroic ma ke kupaa, no laila, o kona aoao akau ma ka Fig.Hoʻokomo ʻia ke ʻano o nā aniani decachromatic i hōʻike ʻia ma ke Kiʻi 3a i kahi adapter 54 mm (kiʻekiʻe) × 58 mm (hohonu) × 8.5 mm (mānoanoa).Ma ka fig.3d, me ka mokuʻāina i hōʻike ʻia ma ka fig.3c, ua hoʻonoho ʻia kahi pahu acrylic piha i ka uahi i mua o kahi ʻano o nā aniani decochromatic, me nā kukui o ka lumi i pio.ʻO ka hopena, ʻike ʻia nā kahawai dichroic ʻeiwa i loko o ka pahu, e puka mai ana mai kahi ʻano o nā aniani decatroic.Loaʻa i kēlā me kēia kahawai māhele kahi ʻāpana keʻa ʻehā me nā ana o 1 × 7 mm, e pili ana i ka nui o ka puka o ka spectrometer ʻeiwa kala hou.Ma ke Kiʻi 3b, ua kau ʻia kahi ʻāpana pepa i mua o ke ʻano o nā aniani dichroic ma ke Kiʻi 3c, a ʻike ʻia ke kiʻi 1 x 7 mm o nā kahawai dichroic ʻeiwa i kuhi ʻia ma luna o ka pepa mai ke ala o ka neʻe ʻana o ka pepa.kahawai.ʻEiwa kala hoʻokaʻawale kahawai ma fig.ʻO 3b a me d he C4, C3, C2, C1, C5, C6, C7, C8 a me C9 mai luna a lalo, hiki ke ʻike ʻia ma nā kiʻi 1 a me 2. 1b a me 2c.ʻIke ʻia lākou i nā kala e pili ana i ko lākou mau nalu.Ma muli o ke kukui keʻokeʻo haʻahaʻa o ka LED (e nānā i ka Fig. S3 Hoʻohui) a me ka naʻau o ke kāmele kala i hoʻohana ʻia no ka hopu ʻana iā C9 (682-699 nm) i ka Fig. Ua nāwaliwali nā kahe ʻokoʻa ʻē aʻe.Pēlā nō, ʻike ʻia ʻo C9 e ka maka ʻōlohelohe.I kēia manawa, ʻo C2 (ke kahawai ʻelua mai luna) ke nānā aku i ka ʻōmaʻomaʻo ma ke Kiʻi 3, akā ʻoi aku ka melemele i ka maka ʻōlohelohe.
Hōʻike ʻia ka hoʻololi ʻana mai ka Figure 3c a i ka d ma ke wikiō Hoʻohui 1. Ma hope koke o ka hele ʻana o ke kukui keʻokeʻo mai ka LED ma o ke aniani aniani decachromatic, ua māhele like ia i ʻeiwa kahawai kala.I ka hopena, ua nalo mālie ka uahi o ka pahu mai luna a lalo, a nalo pū nā pauka ʻeiwa mai luna a lalo.ʻO ka ʻokoʻa, ma ka Video Supplementary 2, i ka wā i hoʻololi ʻia ai ka lōʻihi hawewe o ka hanana ʻana o ke kukui ma luna o ke ʻano o nā aniani decachromatic mai ka lōʻihi a i ka pōkole ma ke kauoha o 690, 671, 650, 632, 610, 589, 568, 550 a me 532 nm. ., Hōʻike wale ʻia nā kahawai mahele like o nā kahawai ʻeiwa ma ke ʻano o C9, C8, C7, C6, C5, C4, C3, C2, a me C1.Hoʻololi ʻia ka waihona wai acrylic e kahi ʻauʻau quartz, a hiki ke ʻike maopopo ʻia nā flakes o kēlā me kēia kahe i hoʻoheheʻe ʻia mai ke ala e piʻi ai i luna.Eia hou, ua hoʻoponopono ʻia ka sub-video 3 i mea e hoʻokani hou ʻia ai ka ʻāpana hoʻololi o ka nalu o ka sub-video 2.ʻO kēia ka hōʻike maikaʻi loa o nā hiʻohiʻona o kahi papa decochromatic o nā aniani.
Hōʻike nā hualoaʻa ma luna nei i ka hana ʻana o ke aniani decachromatic i hana ʻia a i ʻole ka spectrometer ʻeiwa kala hou e like me ka mea i manaʻo ʻia.Hoʻokumu ʻia ka spectrometer ʻeiwa kala hou ma ke kau ʻana i nā aniani decachromatic me nā mea hoʻololi pololei ma luna o ka papa ʻike kiʻi.
ʻO ke kukui kukui me ka lōʻihi o ka hawewe mai 400 a 750 nm, i hoʻokuʻu ʻia e nā helu radiation ʻehā φ50 μm, aia ma 1 mm mau manawa ma ke ala e kū pono ana i ka mokulele o Fig. 2c, i kēlā me kēia noiʻi 31, 34. ʻehā mau lens φ1 mm me ka lōʻihi o 1.4 mm a me ka pitch o 1 mm.ʻEhā mau kahawai i hui pū ʻia (ʻehā C0) i hana ʻia ma ka DP o kahi spectrometer ʻeiwa kala hou, i hoʻokaʻawale ʻia ma 1 mm kaawale.Hoʻokaʻawale nā aniani dichroic i kēlā me kēia kahawai (C0) i ʻeiwa kahawai kala (C1-C9).ʻO ka hopena o nā kahawai 36 (ʻehā pūʻulu o C1-C9) a laila hoʻokomo pololei ʻia i loko o kahi mea ʻike kiʻi CMOS (S) pili pono i kahi ʻano o nā aniani dichroic.ʻO ka hopena, e like me ka mea i hōʻike ʻia ma ka Fig. 5a, ma muli o ka liʻiliʻi o ka ʻokoʻa ʻokoʻa o ke ala optical a me ke ala ʻoi loa pōkole pōkole, ua ʻike ʻia nā kiʻi o nā kahawai 36 a pau i ka manawa like a maopopo me ka nui like.Wahi a ka hiʻohiʻona o lalo (e nānā i ke Kiʻi S4), ʻoi aku ka haʻahaʻa o ke kiʻi o nā hui ʻehā C1, C2 a me C3.He kanakolukumamāono nā kiʻi he 0.57 ± 0.05 mm ka nui (mean ± SD).No laila, ʻo 11.4 ka awelika o ka hoʻonui kiʻi.ʻO ka mokowā kū i waena o nā kiʻi he awelika 1 mm (kokowā like me ka pūnāwai lens) a me ka awelika o ka hoʻopaʻapaʻa ʻāpae he 1.6 mm (kaulike like me ke aniani aniani dichroic).No ka mea ʻoi aku ka liʻiliʻi o ka nui o ke kiʻi ma mua o ka mamao ma waena o nā kiʻi, hiki ke ana ʻia kēlā me kēia kiʻi ma ke kaʻawale (me ka crosstalk haʻahaʻa).I kēia manawa, hōʻike ʻia nā kiʻi o nā kahawai he iwakāluakūmāwalu i hoʻopaʻa ʻia e ka spectrometer ʻehiku kala maʻamau i hoʻohana ʻia i kā mākou noiʻi mua ʻana ma ka Fig. nā aniani ma ke Kiʻi 1a.ʻAʻole paʻakikī nā kiʻi a pau, piʻi ka nui o ke kiʻi mai C1 a i C7.He iwakāluakūmāwalu kiʻi he 0.70 ± 0.19 mm ka nui.No laila, paʻakikī ka mālamaʻana i ka hoʻonā kiʻi kiʻekiʻe ma nā kiʻi a pau.ʻO ka coefficient o ka hoʻololi ʻana (CV) no ka nui o ke kiʻi 28 ma ke Kiʻi 5b he 28%, aʻo ka CV no ka nui o ke kiʻi 36 ma ka Figure 5a i emi iho i 9%.Hōʻike nā hualoaʻa ma luna nei ʻaʻole hoʻonui ka spectrometer ʻeiwa kala hou i ka helu o nā kala i ana ʻia i ka manawa like mai ʻehiku a ʻeiwa, akā loaʻa nō hoʻi kahi hoʻonā kiʻi kiʻekiʻe no kēlā me kēia kala.
Hoʻohālikelike i ka maikaʻi o ke kiʻi māhele i hana ʻia e nā spectrometer maʻamau a hou.(a) ʻEhā pūʻulu o nā kiʻi i hoʻokaʻawale ʻia ʻeiwa kala (C1-C9) i hana ʻia e ka spectrometer ʻeiwa kala hou.(b) ʻEhā pūʻulu o nā kiʻi i hoʻokaʻawale ʻia ʻehiku kala (C1-C7) i hana ʻia me kahi spectrometer ʻehiku kala maʻamau.Hoʻopili ʻia nā Fluxes (C0) me nā lōʻihi hawewe mai 400 a 750 nm mai nā wahi hoʻokuʻu ʻehā a me ka hanana ma kēlā me kēia spectrometer.
Ua loiloi ʻia nā hiʻohiʻona kikoʻī o ka spectrometer ʻeiwa kala a hōʻike ʻia nā hopena loiloi ma ka Figure 6. E hoʻomaopopo ʻo Figure 6a e hōʻike ana i nā hopena like me ka Figure 5a, ʻo ia hoʻi ma nā hawewe o 4 C0 400-750 nm, ʻike ʻia nā kiʻi 36 a pau. (4 pūʻulu C1–C9).ʻOiaʻiʻo, e like me ka mea i hōʻike ʻia ma ke kiʻi 6b–j, ke loaʻa i kēlā me kēia C0 ka nalu kikoʻī o 530, 550, 570, 590, 610, 630, 650, 670, a i ʻole 690 nm, aneane ʻehā wale nō kiʻi pili (ʻehā. ʻike ʻia nā hui C1, C2, C3, C4, C5, C6, C7, C8 a i ʻole C9).Eia nō naʻe, ʻike nāwaliwali loa kekahi o nā kiʻi e pili ana i nā kiʻi pili ʻehā no ka mea ʻo ka C1-C9 hoʻouna kikoʻī i hōʻike ʻia ma ka Fig. 4b ua uhi liʻiliʻi a loaʻa i kēlā me kēia C0 kahi pahu 10 nm ma kahi hawewe kikoʻī e like me ka wehewehe ʻana ma ke ʻano.Kūlike kēia mau hualoaʻa me ka spectra transmission C1-C9 i hōʻike ʻia ma Fig.4b a me nā wikiō hou 2 a me 3. ʻO ia hoʻi, e hana ana ka spectrometer ʻeiwa e like me ka mea i manaʻo ʻia ma muli o nā hopena i hōʻike ʻia ma ka fig.4b.No laila, ua hoʻoholo ʻia ʻo ka puʻunaue ikaika kiʻi C1-C9 ka spectrum o kēlā me kēia C0.
Nā hiʻohiʻona hiʻona o ka spectrometer ʻeiwa kala.Hoʻokumu ka spectrometer ʻeiwa kala hou i ʻehā pūʻulu o nā kiʻi i hoʻokaʻawale ʻia ʻeiwa kala (C1-C9) i ka wā i loaʻa ai i ke kukui hanana (ʻehā C0) ka lōʻihi nalu o (a) 400-750 nm (e like me ka hōʻike ʻia ma ke Kiʻi 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, pakahi.
Ua hoʻohana ʻia ka spectrometer ʻeiwa kala i hoʻomohala ʻia no ka electrophoresis ʻehā-capillary (no nā kikoʻī, e ʻike i nā Mea Hoʻohui)31,34,35.ʻO ka matrix ʻehā-capillary he ʻehā mau capillaries (ka anawaena o waho 360 μm a me ka anawaena o loko 50 μm) aia ma 1 mm mau manawa ma ka pūnaewele irradiation laser.ʻO nā laʻana i loaʻa nā ʻāpana DNA i hoʻopaʻa ʻia me 8 mau kala, ʻo ia hoʻi ʻo FL-6C (dye 1), JOE-6C (dye 2), dR6G (dye 3), TMR-6C (dye 4), CXR-6C (dye 5), TOM- 6C (peu 6), LIZ (peu 7), a me WEN (peu 8) ma ke kaʻina piʻi ʻana o ka lōʻihi hawewe fluorescent, i hoʻokaʻawale ʻia i loko o kēlā me kēia capillaries ʻehā (i kapa ʻia ʻo Cap1, Cap2, Cap3, a me Cap4).Ua hui pū ʻia ka fluorescence laser-induced mai Cap1-Cap4 me kahi ʻano o nā lens ʻehā a hoʻopaʻa pū ʻia me kahi spectrometer ʻeiwa kala.ʻO ka ikaika ikaika o ka ʻeiwa kala (C1-C9) fluorescence i ka wā electrophoresis, ʻo ia hoʻi, kahi electrophoregram ʻeiwa kala o kēlā me kēia capillary, hōʻike ʻia ma Fig. 7a.Loaʻa ka electrophoregram ʻeiwa kala like ma Cap1-Cap4.E like me ka hōʻike ʻana e nā pua Cap1 ma ke Kiʻi 7a, ʻo nā piko ʻewalu ma kēlā me kēia electrophoregram ʻeiwa kala e hōʻike ana i hoʻokahi hoʻoheheʻe fluorescence mai Dye1-Dye8, kēlā me kēia.
ʻO ka helu like ʻana o nā mea pena ʻewalu me ka hoʻohana ʻana i ka spectrometer electrophoresis ʻeiwa kala ʻehā-capillary.(a) ʻEiwa kala (C1-C9) electrophoregram o kēlā me kēia capillary.ʻO nā piko ʻewalu i hōʻike ʻia e nā pua ʻo Cap1 e hōʻike ana i ka hoʻokuʻu ʻana o kēlā me kēia fluorescence o ʻewalu dyes (Dye1-Dye8).Ua pili nā kala o nā pua i nā kala (b) a me (c).(b) Hōʻikeʻike Fluorescence o ʻewalu wai (Dye1-Dye8) no kēlā me kēia capillary.c Electroperograms o ewalu dyes (Dye1-Dye8) no ka capillary.Hōʻike ʻia nā kiʻekiʻe o nā ʻāpana DNA i kapa ʻia ʻo Dye7 e nā pua, a ua hōʻike ʻia ko lākou Cap4 base lōʻihi.
Hōʻike ʻia nā puʻupuʻu ikaika o C1-C9 ma nā piko ʻewalu ma Fig.7b, kēlā me kēia.No ka mea, aia ka C1-C9 a me Dye1-Dye8 i ka hawewe lōʻihi, e hōʻike ana nā puʻunaue ʻewalu ma Fig. 7b i ke kikoʻī fluorescence o Dye1-Dye8 mai ka hema a i ka ʻākau.I loko o kēia haʻawina, ʻike ʻia ʻo Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, a me Dye8 i ka ʻulaʻula, ʻulaʻula, uliuli, cyan, ʻōmaʻomaʻo, melemele, ʻalani, a me ka ʻulaʻula.E hoʻomanaʻo, pili nā kala o nā pua ma ke kiʻi 7a me nā kala kala ma ke kiʻi 7b.ʻO ka ikaika o ka fluorescence C1-C9 no kēlā me kēia kikoʻī ma ke Kiʻi 7b ua hoʻohālikelike ʻia i like ko lākou huina i hoʻokahi.ʻEwalu like ka fluorescence spectra i loaʻa mai Cap1-Cap4.Hiki ke nānā pono i ka uhi ʻana o ka fluorescence ma waena o ka dye 1-dye 8.
E like me ka mea i hōʻike ʻia ma ka Figure 7c, no kēlā me kēia capillary, ua hoʻololi ʻia ka electrophoregram ʻeiwa kala ma ka Figure 7a i kahi electropherogram ewalu-dye e ka loiloi multi-component e pili ana i ka spectra fluorescence ʻewalu ma Figure 7b (e ʻike i nā Mea Hoʻohui no nā kikoʻī).No ka mea, ʻaʻole i hōʻike ʻia ka ʻōniʻoniʻo o ka fluorescence i ke Kiʻi 7a ma ke Kiʻi 7c, hiki ke ʻike a helu ʻia ʻo Dye1-Dye8 i kēlā me kēia manawa, ʻoiai inā ʻokoʻa nā helu o Dye1-Dye8 i ka manawa like.ʻAʻole hiki ke hana ʻia me ka ʻike makaʻu ʻehiku kala kuʻuna31, akā hiki ke hoʻokō ʻia me ka ʻike ʻeiwa kala i kūkulu ʻia.E like me ka mea i hōʻike ʻia e nā pua Cap1 ma Fig. 7c, ʻo nā mea hoʻoheheʻe fluorescent wale nō ʻo Dye3 (uliuli), Dye8 (ʻulaʻula), Dye5 ('ōmaʻomaʻo), Dye4 (cyan), Dye2 (purple), Dye1 (magenta), a me Dye6 (Yellow. ) ʻike ʻia ma ke kaʻina manawa i manaʻo ʻia.No ka hoʻokuʻu ʻia ʻana o ke kala 7 (alani), ma waho aʻe o ka piko hoʻokahi i hōʻike ʻia e ka pua ʻalani, ua ʻike ʻia kekahi mau piko hoʻokahi.ʻO kēia hopena ma muli o ka loaʻa ʻana o nā laʻana i nā kūlana nui, ua kapa ʻia ʻo Dye7 i nā ʻāpana DNA me nā lōʻihi kumu like ʻole.E like me ka hōʻike ʻana ma ke Kiʻi 7c, no Cap4 kēia mau kumu lōʻihi he 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 a me 220 kumu lōʻihi.
ʻO nā hiʻohiʻona nui o ka spectrometer ʻeiwa kala, i hoʻomohala ʻia me ka hoʻohana ʻana i kahi matrix o nā aniani dichroic ʻelua-layer, he liʻiliʻi liʻiliʻi a me ka hoʻolālā maʻalahi.Ma muli o ke ʻano o nā aniani decachromatic i loko o ka adapter i hōʻike ʻia ma ka fig.3c kau pololei ma ka papa ike kiʻi (e nānā i ke kiʻi. S1 a me S2), ʻo ka spectrometer ʻeiwa mau ʻano like me ka adapter, ʻo ia hoʻi 54 × 58 × 8.5 mm.(ka mānoanoa) .ʻO kēia nui liʻiliʻi ʻelua a ʻekolu mau kauoha o ka nui ma mua o nā spectrometers maʻamau e hoʻohana ana i nā gratings a i ʻole prisms.Eia hou, no ka mea, ua hoʻonohonoho ʻia ka spectrometer ʻeiwa ʻulaʻula e like me ka paʻi ʻana o ka mālamalama i ka ʻili o ka mea ʻike kiʻi perpendicularly, hiki ke maʻalahi ka hoʻokaʻawale ʻana i ka lewa no ka spectrometer ʻeiwa kala i loko o nā ʻōnaehana e like me nā microscopes, flow cytometers, a i ʻole nā mea loiloi.ʻO ka mea nānā electrophoresis capillary grating no ka ʻoi aku o ka miniaturization o ka ʻōnaehana.I ka manawa like, ʻo ka nui o nā aniani dichroic he ʻumi a me nā kānana bandpass i hoʻohana ʻia i ka spectrometer ʻeiwa ʻulaʻula he 10 × 1.9 × 0.5 mm a i ʻole 15 × 1.9 × 0.5 mm wale nō.No laila, ʻoi aku ma mua o 100 mau aniani dichroic liʻiliʻi a me nā kānana bandpass, kēlā me kēia, hiki ke ʻoki ʻia mai kahi aniani dichroic a me kahi kānana bandpass 60 mm2.No laila, hiki ke hana ʻia nā aniani decachromatic me ke kumu kūʻai haʻahaʻa.
ʻO kekahi hiʻohiʻona o ka spectrometer ʻeiwa ʻulaʻula ʻo ia kona ʻano hiʻohiʻona maikaʻi loa.ʻO ka mea kūikawā, hiki iā ia ke loaʻa nā kiʻi spectral o nā snapshots, ʻo ia hoʻi, ka loaʻa like ʻana o nā kiʻi me ka ʻike spectral.No kēlā me kēia kiʻi, ua loaʻa kahi spectrum mau me ka lōʻihi o ka hawewe mai 520 a 700 nm a me kahi hoʻonā o 20 nm.Ma nā huaʻōlelo ʻē aʻe, ʻike ʻia nā ʻeiwa kala o ka mālamalama no kēlā me kēia kiʻi, ʻo ia hoʻi, ʻeiwa mau kaha 20 nm e puʻunaue like ana i ka laulā hawewe mai 520 a 700 nm.Ma ka hoʻololi ʻana i nā hiʻohiʻona hiʻohiʻona o ke aniani dichroic a me ka kānana bandpass, hiki ke hoʻoponopono ʻia ka laulā hawewe o nā ʻeiwa a me ka laulā o kēlā me kēia hui.ʻEiwa kala ʻike hiki ke hoʻohana ʻia ʻaʻole wale no nā ana fluorescence me ke kiʻi spectral (e like me ka wehewehe ʻana ma kēia hōʻike), akā no nā noi maʻamau ʻē aʻe me ka hoʻohana ʻana i ke kiʻi spectral.ʻOiai hiki i ke kiʻi hyperspectral ke ʻike i nā haneli o nā kala, ua ʻike ʻia ʻoiai me ka hoʻohaʻahaʻa nui ʻana i ka helu o nā kala ʻike ʻia, hiki ke ʻike ʻia nā mea he nui i ke kahua o ka nānā me ka pololei kūpono no nā noi he nui38,39,40.No ka mea, ʻo ka hoʻonā kikoʻī, ka hoʻonā kikoʻī, a me ka hoʻonā ʻana i ke kino he tradeoff i ka spectral imaging, hoʻemi i ka helu o nā kala hiki ke hoʻomaikaʻi i ka hoʻonā spatial a me ka hoʻonā kino.Hiki iā ia ke hoʻohana i nā spectrometers maʻalahi e like me ka mea i kūkulu ʻia ma kēia haʻawina a hoʻemi hou i ka nui o ka helu.
I loko o kēia noiʻi ʻana, ua helu ʻia nā dyes ʻewalu i ka manawa like e ka hoʻokaʻawale kikoʻī o kā lākou kikoʻī fluorescence overlapping ma muli o ka ʻike ʻana o ʻeiwa kala.Hiki ke helu ʻia a hiki i ʻeiwa i ka manawa like, e noho pū ana i ka manawa a me ka lewa.ʻO kahi pōmaikaʻi kūikawā o ka spectrometer ʻeiwa ʻulaʻula, ʻo ia ke kahe kukui kiʻekiʻe a me ka puka nui (1 × 7 mm).He 92% ka nui o ke kukui mai ka aperture ma kēlā me kēia o nā pae hawewe eiwa.ʻO ka maikaʻi o ka hoʻohana ʻana i ke kukui hanana i ka laulā hawewe mai 520 a 700 nm kokoke i 100%.I loko o kēlā ʻano laulā o nā lōʻihi nalu, ʻaʻole hiki i ka diffraction grating ke hāʻawi i kahi kiʻekiʻe o ka hoʻohana.ʻOiai ke ʻoi aku ka maikaʻi o ka ʻokoʻa o ka hawewe ʻokoʻa ma mua o 90% ma kekahi lōʻihi nalu, i ka piʻi ʻana o ka ʻokoʻa ma waena o kēlā lōʻihi nalu a me kekahi lōʻihi hawewe, e emi ana ka maikaʻi o ka diffraction ma kekahi lōʻihi nalu41.Hiki ke hoʻonui ʻia ka laulā puka e kū pono ana i ke kuhikuhi o ka mokulele ma Fig. 2c mai ka 7 mm a hiki i ka laula o ka ʻike kiʻi, e like me ke ʻano o ka ʻike kiʻi i hoʻohana ʻia ma kēia haʻawina, ma ka hoʻololi iki ʻana i ka papa decamer.
ʻAʻole hiki ke hoʻohana ʻia ka spectrometer ʻeiwa ʻulaʻula no ka electrophoresis capillary, e like me ka hōʻike ʻana ma kēia haʻawina, akā no nā kumu ʻē aʻe.No ka laʻana, e like me ka mea i hōʻike ʻia ma ke kiʻi ma lalo nei, hiki ke hoʻohana ʻia kahi spectrometer ʻeiwa kala i kahi microscope fluorescence.Hōʻike ʻia ka mokulele o ka hāpana ma ka ʻike kiʻi o ka spectrometer ʻeiwa kala ma o ka pahuhopu 10x.ʻO 200 mm ka mamao ʻike maka ma waena o ka lens a me ka ʻike kiʻi, ʻoiai ʻo ka mamao ma waena o ka ili i hana ʻia o ka spectrometer ʻeiwa kala a me ka ʻike kiʻi he 12 mm wale nō.No laila, ua ʻoki ʻia ke kiʻi e like me ka nui o ka aperture (1 × 7 mm) ma ka mokulele o ka hanana a māhele ʻia i ʻeiwa kiʻi kala.ʻO ia hoʻi, hiki ke kiʻi ʻia ke kiʻi spectral o kahi kiʻi ʻeiwa kala ma kahi 0.1 × 0.7 mm wahi i ka mokulele hāpana.Eia hou, hiki ke kiʻi i kahi kiʻi spectral ʻeiwa kala o kahi ʻāpana nui ma ka mokulele hāpana ma ka nānā ʻana i ka hāpana e pili ana i ka pahuhopu ma ka ʻaoʻao ākea ma Fig. 2c.
ʻO nā ʻāpana hoʻonohonoho aniani decachromatic, ʻo ia hoʻi ʻo M1-M9 a me BP, i hana maʻamau e Asahi Spectra Co., Ltd. me ka hoʻohana ʻana i nā ʻano ua maʻamau.Ua hoʻopili ʻia nā mea dielectric multilayer ma luna o nā pā quartz he ʻumi 60 × 60 mm ka nui a me 0.5 mm mānoanoa, e hālāwai ana i kēia mau koi: M1: IA = 45 °, R ≥ 90% ma 520–590 nm, Tave ≥ 90% ma 610– 610 nm.700 nm, M2: IA = 45°, R ≥ 90% ma 520–530 nm, Tave ≥ 90% ma 550–600 nm, M3: IA = 45°, R ≥ 90% ma 540–550 nm, Tave ≥ 90 % ma 570–600 nm, M4: IA = 45°, R ≥ 90% ma 560–570 nm, Tave ≥ 90% ma 590–600 nm, M5: IA = 45°, R ≥ 98% ma 580–600 nm , R ≥ 98% ma 680–700 nm, M6: IA = 45°, Tave ≥ 90% ma 600–610 nm, R ≥ 90% ma 630–700 nm, M7: IA = 45°, R ≥ 90% ma 620–630 nm, Taw ≥ 90% ma 650–700 nm, M8: IA = 45°, R ≥ 90% ma 640–650 nm, Taw ≥ 90% ma 670–700 nm, M9: IA = 45°, R ≥ 90% ma 650-670 nm, Tave ≥ 90% ma 690-700 nm, BP: IA = 0°, T ≤ 0.01% ma 505 nm, Tave ≥ 95% ma 530-690 nm ma 530 nm T ≥ ma -690 nm a me T ≤ 1% ma 725-750 nm, kahi o IA, T, Tave, a me R ka huina o ka hanana, transmittance, average transmittance, a me unpolarized malamalama reflectance.
ʻO ke kukui keʻokeʻo (C0) me ka laulā hawewe o 400–750 nm i hoʻokuʻu ʻia e kahi kumu kukui LED (AS 3000, AS ONE CORPORATION) i hui pū ʻia a kū pololei ma ka DP o nā aniani dichroic.Hōʻike ʻia ke ʻano kukui keʻokeʻo o nā LED ma ke Kiʻi S3.E kau i kahi pahu pahu acrylic (nā ana 150 × 150 × 30 mm) i mua pono o ke aniani aniani decamera, e kū pono ana i ka PSU.ʻO ka uahi i hana ʻia i ka wā i hoʻokomo ʻia ai ka hau maloʻo i loko o ka wai, a laila ninini ʻia i loko o kahi pahu acrylic e nānā i nā kahawai ʻeiwa ʻeiwa C1-C9 e puka mai ana mai ke ʻano o nā aniani decachromatic.
ʻO kahi ʻē aʻe, hele ka kukui keʻokeʻo collimated (C0) i kahi kānana ma mua o ke komo ʻana i ka DP.ʻO nā kānana he mau kānana kūʻokoʻa kūʻokoʻa me ka 0.6 optical density.A laila e hoʻohana i kahi kānana kaʻa (FW212C, FW212C, Thorlabs).ʻO ka hope, hoʻohuli hou i ka kānana ND.Hoʻopili nā bandwidth o nā kānana bandpass ʻeiwa me C9, C8, C7, C6, C5, C4, C3, C2 a me C1.Ua waiho ʻia kahi cell quartz me nā ana o loko o 40 (lōʻihi optical) x 42.5 (kiʻekiʻe) x 10 mm (ākea) i mua o kahi ʻano o nā aniani decochromatic, e kū pono ana i ka BP.Hāʻawi ʻia ka uahi ma o ka paipu i loko o ka cell quartz no ka mālama ʻana i ka neʻe ʻana o ka uahi i loko o ka cell quartz e ʻike i nā kahawai ʻeiwa C1-C9 i hoʻokaʻawale ʻia mai ke aniani aniani decachromatic.
Ua kiʻi ʻia kahi wikiō o ke kahawai māmā ʻeiwa ʻulaʻula e puka mai ana mai ke ʻano o nā aniani decanic i ke ʻano manawa-lapse ma ka iPhone XS.E kiʻi i nā kiʻi o ka hiʻohiʻona ma 1 fps a hōʻuluʻulu i nā kiʻi e hana i wikiō ma 30 fps (no ke wikiō koho 1) a i ʻole 24 fps (no nā wikiō koho 2 a me 3).
E kau i 50 µm mānoanoa pā kila kuhiliʻi (me ʻehā 50 µm puka anawaena ma 1 mm kaawale) ma ka pā diffusion.Hoʻopili ʻia ka mālamalama me ka lōʻihi nalu o 400-750 nm ma luna o ka pā diffuser, loaʻa ma ka hāʻawi ʻana i ke kukui mai kahi kukui halogen ma o kahi kānana hoʻouna pōkole me ka lōʻihi o ka hawewe o 700 nm.Hōʻike ʻia ke kiko uila ma ke Kiʻi S4.ʻO kahi ʻē aʻe, hele pū ka mālamalama i kekahi o nā kānana bandpass 10 nm i kikowaena ma 530, 550, 570, 590, 610, 630, 650, 670 a me 690 nm a paʻi i ka pā diffuser.ʻO ka hopena, ua hoʻokumu ʻia nā helu radiation ʻehā me ke anawaena o φ50 μm a me nā lōʻihi hawewe like ʻole ma luna o kahi pā kila kila e kū pono ana i ka pā diffuser.
Hoʻokomo ʻia kahi ʻāpana ʻehā-capillary me nā lens ʻehā ma kahi spectrometer ʻeiwa kala e like me ka hōʻike ʻana ma nā Kiʻi 1 a me 2. C1 a me C2.Ua like nā capillaries ʻehā a me nā lens ʻehā me nā haʻawina mua31,34.ʻO kahi kukuna laser me ka lōʻihi o ka nalu o 505 nm a me ka mana o 15 mW e hoʻomālamalama ʻia i ka manawa like a me ke kaulike mai ka ʻaoʻao a hiki i nā wahi hoʻokuʻu o nā capillaries ʻehā.Hoʻopili ʻia ka fluorescence i hoʻokuʻu ʻia e kēlā me kēia wahi hoʻokuʻu ʻia e ka lens e pili ana a hoʻokaʻawale ʻia i ʻeiwa mau kahawai waihoʻoluʻu e ke ʻano o nā aniani decachromatic.A laila, hoʻokomo pololei ʻia nā kahawai he 36 i loko o kahi mea ʻike kiʻi CMOS (C11440–52U, Hamamatsu Photonics K·K.), a ua hoʻopaʻa ʻia ko lākou mau kiʻi i ka manawa like.
ʻO ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Applied Biosystems), 4 µl GeneScan™ 600 LIZ™ dye i hui ʻia no kēlā me kēia capillary ma o ka hui ʻana i 1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), 1 µl mix size standard.v2.0 (Thermo Fisher Scientific) a me 14 µl wai.ʻO ka PowerPlex® 6C Matrix Standard he ʻeono mau ʻāpana DNA i hōʻailona ʻia me ʻeono mau kala: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, a me WEN, ma ke ʻano o ka lōʻihi hawewe.ʻAʻole hōʻike ʻia ka lōʻihi kumu o kēia mau ʻāpana DNA, akā ʻike ʻia ke kaʻina lōʻihi kumu o nā ʻāpana DNA i kapa ʻia me WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C a me TOM-6C.Aia i loko o ka ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit kahi ʻāpana DNA i hōʻailona ʻia me ka dR6G dye.ʻAʻole i hōʻike ʻia ka lōʻihi o nā kumu o nā ʻāpana DNA.Aia ʻo GeneScan™ 600 LIZ™ Dye Size Standard v2.0 me 36 LIZ-labeled ʻāpana DNA.ʻO nā lōʻihi kumu o kēia mau ʻāpana DNA he 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 314, 30 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 a me 600 kumu.Hoʻopau ʻia nā laʻana ma 94 ° C no 3 mau minuke, a laila hoʻomaha ʻia ma ka hau no 5 mau minuke.Hoʻokomo ʻia nā laʻana i loko o kēlā me kēia capillary ma 26 V/cm no 9 s a hoʻokaʻawale ʻia i kēlā me kēia capillary i hoʻopiha ʻia me kahi solution polymer POP-7™ (Thermo Fisher Scientific) me ka lōʻihi o 36 knm a me ka volta o 181 V/cm a me kahi. ka huina o 60°.MAI.
ʻO nā ʻikepili a pau i loaʻa a i ʻike ʻia paha i loko o ka papa o kēia haʻawina i loko o kēia ʻatikala i paʻi ʻia a me kāna ʻike hou.Loaʻa nā ʻikepili ʻē aʻe e pili ana i kēia noiʻi mai nā mea kākau ma muli o ke noi kūpono.
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