igwe anaghị agba nchara 321 8 * 1.2 ọkpọkọ ọkpọkọ maka okpomọkụ Exchanger

Ọdụdọ Capillary

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Emepụtara ultra-compact (54 × 58 × 8.5 mm) na oghere obosara (1 × 7 mm) spectrometer agba agba iteghete, “gbawara abụọ” site n'ụdị enyo dichroic iri, nke ejiri mee ihe onyonyo ozugbo.Ọkụ ọkụ ahụ merenụ nke nwere akụkụ obe pere mpe karịa oghere oghere na-ekewa n'ime warara na-aga n'ihu 20nm n'obosara yana fluxes agba itoolu nwere ogologo wavelength nke 530, 550, 570, 590, 610, 630, 650, 670 na 690 nm.A na-atụ ihe onyonyo iyi agba agba itoolu n'otu oge nke ọma site na ihe mmetụta onyonyo.N'adịghị ka ihe eji enyo enyo dichroic a na-emekarị, ihe eji emepụta ihe na-emepụta ihe na-emepụta ihe nwere nhazi abụọ pụrụ iche, nke ọ bụghị nanị na-eme ka ọnụ ọgụgụ nke agba nke nwere ike ịlele n'otu oge, ma na-eme ka mkpebi oyiyi dịkwuo mma maka iyi agba agba ọ bụla.A na-eji spectrometer agba agba itoolu emepụtara maka electrophoresis nke capillary anọ.Ntụle ọnụ ọgụgụ nke otu oge nke agba agba asatọ na-akwaga n'otu oge na capillary ọ bụla na-eji fluorescence nwere agba laser agba itoolu.Ebe ọ bụ na spectrometer agba agba itoolu abụghị naanị ultra-obere na ọnụ ala, kamakwa ọ nwere nnukwu ọkụ na-egbuke egbuke na mkpebi zuru oke maka ọtụtụ ngwa eserese eserese, enwere ike iji ya mee ihe n'ọtụtụ ebe.
Hyperspectral na multispectral imaging aghọwo akụkụ dị mkpa nke astronomy2, remote sensing for Earth observation3,4, Food and water quality control5,6, art conservation and archeology7, forensics8, surgery9, biomedical analysis and diagnostics10,11 etc. Ubi 1 Teknụzụ dị mkpa dị mkpa. ,12,13.A na-ekewa ụzọ maka ịtụ ụdị ọkụ dị iche iche na-ewepụta site n'akụkụ ọ bụla nke na-ekupụta n'ubi a na-ahụ anya na (1) nyocha ihe ("broom") , (3) ogologo na-enyocha ebili mmiri19,20,21 na (4) oyiyi22,23,24,25.N'ihe gbasara ụzọ ndị a niile, mkpebi gbasara ohere, nhụsianya nhụsianya na mkpebi nwa oge nwere mmekọrịta ahia9,10,12,26.Na mgbakwunye, mmepụta ọkụ na-enwe mmetụta dị ukwuu na nghọta, ya bụ nrịbama-na-mkpọtụ na spectral imaging26.Mmụgharị na-enwu enwu, ya bụ, arụmọrụ nke iji ọkụ, dabara kpọmkwem na nha nke nha nha ọkụ nke ọ bụla na-enwupụta ìhè n'otu oge na ngụkọta nke ìhè nke ogologo ikuku tụrụ.Otu (4) bụ usoro kwesịrị ekwesị mgbe ike ma ọ bụ ụdị dị iche iche nke ìhè na-apụta site n'oge ọ bụla na-ekupụta na-agbanwe n'oge ma ọ bụ mgbe ọnọdụ nke ọ bụla na-ekupụta na-agbanwe ka oge na-aga n'ihi na a na-atụle ụdị ọkụ dị iche iche nke ihe niile na-ebupụta n'otu oge.24.
Ọtụtụ n'ime ụzọ ndị a dị n'elu jikọtara ya na nnukwu, mgbagwoju anya na / ma ọ bụ ọnụ ahịa spectrometers na-eji 18 gratings ma ọ bụ 14, 16, 22, 23 prisms maka klaasị (1), (2) na (4) ma ọ bụ 20, 21 diski nzacha, ihe nzacha mmiri. .Ihe nzacha kristal tunable (LCTF)25 ma ọ bụ ihe nzacha acousto-optic tunable (AOTF)19 nke otu (3).N'ụzọ dị iche, ụdị (4) spectrometer multi-mirror dị obere ma dị ọnụ ala n'ihi nhazi ha dị mfe27,28,29,30.Tụkwasị na nke ahụ, ha nwere nnukwu ọkụ na-egbuke egbuke n'ihi na ọkụ na-ekerịta site na enyo dichroic ọ bụla (ya bụ, a na-ebufe na nke na-egosipụta ìhè nke ìhè merenụ na enyo dichroic ọ bụla) na-eji ya eme ihe n'ụzọ zuru ezu na-aga n'ihu.Agbanyeghị, ọnụ ọgụgụ nke eriri ogologo ogologo (ya bụ agba) nke a ga-atụrịrị n'otu oge bụ ihe dị ka anọ.
A na-ejikarị ihe ngosi nke na-adabere na nchọpụta fluorescence maka nyocha multiplex na nchọpụta biomedical na nchọpụta 10, 13.Na multiplexing, ebe ọ bụ na ọtụtụ nyocha (dịka ọmụmaatụ, DNA ma ọ bụ protein dị iche iche) ka ejiri akwa fluorescent dị iche iche , onye nyocha ọ bụla dị na ebe ọ bụla na-epupụta n'ọhụụ na-enyocha site na iji nyocha multicomponent.32 na-akụrisị ụdị fluorescence achọpụtara nke isi ihe ọ bụla na-ewepụta.N'oge usoro a, agba dị iche iche, nke ọ bụla na-emepụta ihe dị iche iche, nwere ike ịchakọta, ya bụ, ibikọ ọnụ na mbara igwe na oge.Ka ọ dị ugbu a, ọnụ ọgụgụ kachasị nke agba agba nke nwere ike inwe obi ụtọ site na otu ọkụ ọkụ laser bụ asatọ33.Ekpebighị oke a dị elu site na mkpebi nlegharị anya (ya bụ, ọnụ ọgụgụ agba), mana site na obosara nke ụdị fluorescence spectrum (≥50 nm) yana ọnụọgụ dye Stokes shift (≤200 nm) na FRET (iji FRET)10 .Otú ọ dị, ọnụ ọgụgụ nke agba ga-adị ukwuu karịa ma ọ bụ hà nhata na ọnụ ọgụgụ nke e ji esiji ákwà iji wepụ ihe dị iche iche nke agba agba agwakọta31,32.Ya mere, ọ dị mkpa ịbawanye ọnụ ọgụgụ nke agba a tụrụ n'otu oge ruo asatọ ma ọ bụ karịa.
N'oge na-adịbeghị anya, e mepụtala ultra-compact heptachroic spectrometer (na-eji ọtụtụ enyo heptychroic na ihe mmetụta onyonyo iji tụọ fluorescent fluxes anọ).The spectrometer bụ iwu abụọ ma ọ bụ atọ nke ịdị ukwuu karịa spectrometers na-eji gratings ma ọ bụ prisms34,35.Agbanyeghị, ọ siri ike idowe ihe karịrị enyo dichroic asaa na spectrometer ma tụọ ihe karịrị agba asaa36,37.Site na mmụba nke enyo dichroic, ihe dị iche iche dị n'ogologo nke ụzọ anya anya nke ọkụ ọkụ dichroic na-abawanye, ọ na-esikwa ike igosipụta ọkụ ọkụ niile n'otu ụgbọ elu sensory.Ogologo ụzọ anya kachasị ogologo nke ọkụ ọkụ na-abawanye, ya mere obosara nke oghere spectrometer (ya bụ obosara kachasị nke ìhè nke nyocha nke spectrometer) na-ebelata.
Na nzaghachi nye nsogbu ndị a dị n'elu, e mepụtara spectrometer dị agba agba agba itoolu nke nwere akwa abụọ "dichroic" decachromatic mirror array na ihe mmetụta ihe oyiyi maka ihe oyiyi ngwa ngwa ngwa ngwa [ụdị (4)].Tụnyere spectrometers gara aga, spectrometer mepere emepe nwere obere ihe dị iche na ogologo ụzọ anya kachasị na ogologo ụzọ anya kacha nta.A na-etinye ya na electrophoresis nke capillary anọ iji chọpụta fluorescence agba itoolu nke laser na-eme ka ọ bụrụ ọnụ ọgụgụ nke njem n'otu oge nke agba asatọ na capillary ọ bụla.Ebe ọ bụ na spectrometer emepụtara abụghị naanị ultra-obere na ọnụ ala, kamakwa ọ nwere nnukwu ọkụ na-enwu ọkụ yana mkpebi nlegharị anya zuru oke maka ọtụtụ ngwa onyonyo onyonyo, enwere ike iji ya na mpaghara dị iche iche.
A na-egosi spectrometer agba agba ochie na fig.1a.Nhazi ya na-esote nke gara aga ultra-small asaa spectrometer 31. Ọ mejupụtara enyo dichroic itoolu nke a na-ahazi n'usoro n'akụkụ 45 ° n'aka nri, na ihe mmetụta oyiyi (S) dị n'elu enyo dichroic itoolu.A na-ekewa ìhè nke na-esi n'okpuru (C0) na-abanye n'usoro nke enyo dichroic itoolu n'ime ọkụ itoolu na-agbago (C1, C2, C3, C4, C5, C6, C7, C8 na C9).A na-enye mmiri iyi agba agba itoolu niile ozugbo na ihe mmetụta onyonyo wee chọpụta n'otu oge.N'ime ọmụmụ ihe a, C1, C2, C3, C4, C5, C6, C7, C8, na C9 bụ n'usoro ogologo ogologo ma na-anọchi anya magenta, violet, blue, cyan, green, yellow, orange, red-orange, na nọchitere anya ya. uhie, n'otu n'otu.Ọ bụ ezie na a na-eji aha agba ndị a mee ihe n'akwụkwọ a, dịka e gosiri na foto nke 3, n'ihi na ha dị iche na agba nke anya mmadụ na-ahụ.
Eserese nke spectrometer agba agba ọhụrụ na nke ọhụrụ.(a) spectrometer agba agba itoolu nwere ọtụtụ enyo dichroic itoolu.(b) Ọhụrụ spectrometer agba agba itoolu nwere nhazi enyo dichroic nwere oyi akwa abụọ.E kewara nfegharị ọkụ ahụ C0 n'ime ọkụ ọkụ nwere agba itoolu C1-C9 wee hụ ya site na ihe mmetụta onyonyo S.
Igwe spectrometer agba agba agba itoolu ọhụrụ mepere emepe nwere grating mirror dichroic nke nwere okpukpu abụọ na ihe mmetụta onyonyo, dị ka egosiri na Fig. 1b.N'ọkwa dị ala, enyo dichroic ise na-ehulata 45° n'aka nri, kwekọọ n'aka nri site na etiti n'usoro nke decamers.N'elu ọkwa dị elu, a na-atụgharị enyo dichroic ise ọzọ 45 ° n'aka ekpe ma dị site na etiti ruo n'aka ekpe.Ngosipụta dichroic nke dị n'aka ekpe nke oyi akwa dị ala na enyo dichroic aka nri nke elu oyi akwa na-ekpuchi ibe ya.A na-ekewa ọkụ ọkụ ahụ merenụ (C0) site n'okpuru ebe a gaa na chromatic fluxes anọ na-apụ apụ (C1-C4) site na enyo dichroic ise n'aka nri na fluxes chromatic ise (C5-C4) site na enyo dichroic ise n'aka ekpe C9).Dị ka spectrometer agba agba itoolu, a na-agbanye mmiri iyi agba agba itoolu niile ozugbo n'ime ihe mmetụta onyonyo (S) wee chọpụta n'otu oge.N'ịtụle ọnụ ọgụgụ 1a na 1b, mmadụ nwere ike ịhụ na n'ihe gbasara spectrometer ọhụrụ agba agba itoolu, ma ọdịiche kachasị elu na ogologo ụzọ anya anya ogologo nke agba agba agba itoolu na-ebelata.
E gosipụtara nkọwa zuru ezu nke ihe owuwu nke ultra-obere okpukpu abụọ dichroic mirror array 29 mm (obosara) × 31 mm (omi) × 6 mm (ịdị elu) na eserese 2. Ihe ngosi enyo dichroic decimal nwere enyo dichroic ise n'aka nri. (M1-M5) na enyo dichroic ise dị n'aka ekpe (M6-M9 na M5 ọzọ), enyo dichroic ọ bụla na-edozi n'elu ihe nkwado aluminum.A na-akwagharị enyo niile dichroic iji kwụọ ụgwọ maka mbugharị n'otu n'otu n'ihi ntugharị nke mgbaba na enyo.N'okpuru M1, a na-edozi ihe nzacha band-pass (BP).Akụkụ M1 na BP bụ 10mm (akụkụ ogologo) x 1.9mm (akụkụ dị mkpirikpi) x 0.5mm (ọkpụrụkpụ).Akụkụ nke enyo dichroic fọdụrụnụ bụ 15 mm × 1.9 mm × 0.5 mm.Oghere matriks dị n'etiti M1 na M2 bụ 1.7 mm, ebe matrix pitch nke enyo dichroic ndị ọzọ bụ 1.6 mm.Na fig.2c na-ejikọta ihe ọkụ ọkụ C0 merenụ na fluxes agba agba itoolu C1-C9, nke enyo de-chamber kewapụrụ.
Mwube nke matriks mirror dichroic oyi akwa abụọ.(a) Nlele anya na (b) nleba anya n'ofe akụkụ nke ụdị mirror dichroic nke nwere oyi akwa abụọ (akụkụ 29 mm x 31 mm x 6 mm).Ọ nwere enyo dichroic ise (M1-M5) nke dị na oyi akwa ala, enyo dichroic ise (M6-M9 na M5 ọzọ) dị n'elu oyi akwa, yana eriri bandpass (BP) dị n'okpuru M1.(c) Nleba anya n'ofe akụkụ na ntụzịaka kwụ ọtọ, yana C0 na C1-C9 na-atụgharị.
The obosara nke oghere na kehoraizin direction, gosiri site obosara C0 na fig. 2, c, bụ 1 mm, na na ntụziaka perpendicular na ụgbọ elu nke Fig. 2, c, nyere site imewe nke aluminum bracket. - 7 mm.Ya bụ, spectrometer ọhụrụ agba itoolu nwere nnukwu oghere nke 1 mm × 7 mm.Ụzọ anya nke C4 bụ nke kachasị ogologo n'etiti C1-C9, na ụzọ anya nke C4 n'ime nhazi mirror dichroic, n'ihi nha dị n'elu ultra-obere (29 mm × 31 mm × 6 mm), bụ 12 mm.N'otu oge ahụ, ogologo ụzọ anya nke C5 bụ nke kacha nso n'etiti C1-C9, na ogologo ụzọ anya nke C5 bụ 5.7mm.Ya mere, ihe kachasị dị iche na ogologo ụzọ anya bụ 6.3 mm.A na-edozi ogologo ụzọ anya n'elu maka ogologo ụzọ anya maka nnyefe anya nke M1-M9 na BP (site na quartz).
A na-agbakọ ihe dị iche iche nke М1-М9 na VR ka fluxes С1, С2, С3, С4, С5, С6, С7, С8 na С9 nọ n'ogo ogologo 520-540, 540-560, 560-580, 560-8580. – 600, 600–620, 620–640, 640–660, 660–680, na 680–700 nm n’otu n’otu.
E gosipụtara foto nke matriks arụpụtara nke enyo decachromatic na Fig. 3a.A na-ejikọta M1-M9 na BP na mkpọda 45 ° na kwụ ọtọ nke nkwado aluminom, n'otu n'otu, ebe M1 na BP na-ezobe na azụ nke ọnụ ọgụgụ ahụ.
Mmepụta nke enyo decan dị iche iche na ngosipụta ya.(a) Ọtụtụ enyo decachromatic emepụtara.(b) Ihe onyonyo 1 mm × 7 mm gbawara agbawa nke agba ite iteghete n'elu mpempe akwụkwọ edobere n'ihu ọtụtụ enyo decachromatic na azụ nwere ọkụ na-acha ọcha.(c) Ọtụtụ enyo decochromatic ejiri ọkụ na-acha ọcha na-enwu n'azụ.(d) iyi nke na-agbawa agba itoolu nke na-esite n'usoro enyo decane, nke a na-ahụ site n'itinye akpa acrylic jupụtara na anwụrụ ọkụ n'ihu n'usoro enyo decane na c ma na-agbachi ọnụ ụlọ ahụ.
Atụtara ụdị mgbasa ozi nke M1-M9 C0 n'akụkụ ihe omume nke 45 ° na ụdị nnyefe a tụrụ atụ nke BP C0 n'akụkụ akụkụ nke 0 Celsius ka egosiri na fig.4a.E gosipụtara ụdị mgbasa ozi nke C1-C9 metụtara C0 na fig.4b.A gbakọrọ ụdịdị ndị a site na ụdịdị dị na fig.4a dị ka ụzọ anya anya C1-C9 na Fig. 4a.1b na 2c.Dịka ọmụmaatụ, 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)], ebe TS (X) na [1 - TS (X)] bụ nnyefe na ntụgharị uche nke X, n'otu n'otu.Dị ka egosiri na eserese 4b, bandwit (bandwidth ≥50%) nke C1, C2, C3, C4, C5, C6, C7, C8 na C9 bụ 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 na 682-699 nm.Nsonaazụ ndị a dabara na oke ndị emepụtara.Na mgbakwunye, arụmọrụ ojiji nke ọkụ C0 dị elu, ya bụ, nkezi kacha C1-C9 nnyefe ọkụ bụ 92%.
Ihe nlegharị anya nke enyo dichroic na mgbawa agba agba ite iteghete.(a) Atụle ụdị mgbasa ozi nke M1-M9 na 45° omume na BP na 0°.(b) Ụdị nnyefe nke C1–C9 metụtara C0 gbakọrọ site na (a).
Na fig.3c, n'usoro nke enyo dichroic dị na kwụ ọtọ, nke mere na akụkụ aka nri ya na Fig. 3a bụ n'akụkụ elu ma na-acha ọcha ọkụ nke collimated LED (C0) bụ azụ azụ.N'usoro nke enyo decachromatic nke egosiri na eserese 3a ka etinyere na nkwụnye 54 mm (ịdị elu) × 58 mm (omimi) × 8.5 mm (ọkpụrụkpụ).Na fig.3d, na mgbakwunye na steeti egosiri na fig.3c, a na-etinye tankị acrylic jupụtara na anwụrụ ọkụ n'ihu ọtụtụ enyo decochromatic, na-agbanye ọkụ n'ime ụlọ ahụ.N'ihi ya, a na-ahụ iyi dichroic itoolu n'ime tank ahụ, na-esite n'ọtụtụ enyo decachromatic.iyi nke ọ bụla kewara nwere akụkụ obe akụkụ anọ nwere akụkụ nke 1 × 7 mm, nke dabara na nha oghere nke spectrometer agba ọhụrụ itoolu.Na eserese 3b, a na-edobe akwụkwọ mpempe akwụkwọ n'ihu nhazi nke enyo dichroic dị na eserese 3c, na ihe oyiyi 1 x 7 mm nke iyi dichroic itoolu nke a na-eme n'elu akwụkwọ ahụ na-ahụ site na ntụziaka nke mmegharị akwụkwọ.iyi.Na agba nkewa nke itoolu na fig.3b na d bụ ​​C4, C3, C2, C1, C5, C6, C7, C8 na C9 site n'elu ruo na ala, nke nwekwara ike ịhụ na ọnụ ọgụgụ 1 na 2. 1b na 2c.A na-ahụ ha na agba dabara na ogologo ogologo ogologo ha.N'ihi obere ọkụ ọkụ na-acha ọcha nke LED (lee Mgbakwunye Fig. S3) na mmetụta nke igwefoto agba ejiri jide C9 (682-699 nm) na Fig. Ndị ọzọ na-agbawa agbawa adịghị ike.N'otu aka ahụ, a na-ahụ C9 anya nke ọma.Ka ọ dị ugbu a, C2 (iyi nke abụọ si n'elu) na-ele anya akwụkwọ ndụ akwụkwọ ndụ na Figure 3, mana ọ na-ele anya na-acha odo odo na anya gba ọtọ.
E gosipụtara mgbanwe site na eserese 3c ruo d na vidiyo mgbakwunye 1. Ozugbo ọkụ na-acha ọcha sitere na LED gafere n'usoro enyo decachromatic, ọ na-ekewa n'otu oge gaa na iyi agba agba itoolu.N'ikpeazụ, anwụrụ ọkụ dị na vat ahụ ji nwayọọ nwayọọ na-esi n'elu ruo na ala, nke mere na ntụ ntụ itoolu ahụ na-apụkwa n'anya site n'elu ruo ala.N'ụzọ dị iche, na mgbakwunye vidiyo 2, mgbe ogologo ebili mmiri nke mgbagharị ọkụ n'ụdị enyo decachromatic gbanwere site na ogologo gaa na nkenke n'usoro nke 690, 671, 650, 632, 610, 589, 568, 550 na 532 nm. ., Naanị iyi nke kewara ekewa kwekọrọ na iyi nke ite ite iteghete n'usoro nke C9, C8, C7, C6, C5, C4, C3, C2, na C1 ka egosipụtara.A na-eji ọdọ mmiri quartz dochie ebe nchekwa acrylic, na flakes nke ọ bụla shunted eruba nwere ike ịhụ nke ọma site na mgbada ugwu.Na mgbakwunye, a na-edezi sub-video 3 nke mere na a na-emegharị akụkụ nke obere vidiyo nke 2 n'ogologo.Nke a bụ okwu kachasị mma nke njirimara nke enyo decochromatic.
Nsonaazụ ndị a dị n'elu na-egosi na nhazi mirror decachromatic arụpụtara ma ọ bụ spectrometer ọhụrụ agba agba itoolu na-arụ ọrụ dịka e bu n'obi.A na-emepụta spectrometer ọhụrụ agba agba itoolu site n'ịkwanye ọtụtụ enyo decachromatic nwere ihe nkwụnye ozugbo na bọọdụ ihe mmetụta onyonyo.
Luminous flux na a wavelength nso si 400 ka 750 nm, emitted site anọ radieshon isi φ50 μm, emi odude ke 1 mm etiti oge na ntụziaka perpendicular ka ụgbọ elu nke Fig. 2c, karị Researches 31, 34. The anọ oghere n'usoro mejupụtara nke. lenses anọ φ1 mm na ogologo nke 1.4 mm na pitch nke 1 mm.Iyi anọ agbakọtara (C0 anọ) bụ ihe mere na DP nke ihe nlere anya agba agba ọhụrụ itoolu, nke etinyere na etiti 1 mm.Ọtụtụ enyo dichroic na-ekewa iyi ọ bụla (C0) gaa na iyi agba agba itoolu (C1-C9).A na-agbanye iyi iyi 36 (usoro anọ nke C1-C9) ozugbo n'ime ihe mmetụta onyonyo CMOS (S) jikọtara ya na ọtụtụ enyo dichroic.N'ihi ya, dị ka e gosiri na Fig. 5a, n'ihi ntakịrị ihe dị iche iche ụzọ anya anya na ụzọ dị mkpirikpi, a na-achọpụta ihe oyiyi nke iyi iyi 36 niile n'otu oge na n'ụzọ doro anya na otu nha.Dika ihe nlere anya nke ala (lee ihe onyonyo S4), ike onyonyo nke otu anọ C1, C2 na C3 dị obere.Onyonyo iri atọ na isii bụ 0.57 ± 0.05 mm n'ogo (pụtara ± SD).Ya mere, mmụba onyonyo a ruru 11.4.Oghere kwụ ọtọ n'etiti onyonyo ruru 1 mm (otu oghere dị ka oghere oghere) yana oghere kwụ ọtọ bụ 1.6 mm (otu oghere dị ka nhazi enyo dichroic).N'ihi na nha onyonyo dị ntakịrị karịa ebe dị n'etiti onyonyo, enwere ike tụọ onyonyo ọ bụla n'onwe ya (na obere okwu mkparịta ụka).Ka ọ dị ugbu a, a na-egosi ihe oyiyi iyi iyi iri abụọ na asatọ nke spectrometer agba asaa nke a na-eji eme ihe n'ọmụmụ ihe anyị gara aga na-egosi na Fig. 5 B. A na-emepụta ihe oyiyi nke enyo dichroic asaa site n'iwepụ enyo abụọ dichroic ziri ezi site n'usoro nke dichroic itoolu. enyo dị na eserese 1a.Ọ bụghị onyonyo niile dị nkọ, nha onyonyo na-abawanye site na C1 ruo C7.Onyonyo iri abụọ na asatọ bụ 0.70 ± 0.19 mm n'ogo.Ya mere, ọ na-esiri ike ịnọgide na-enwe mkpebi dị elu na ihe oyiyi niile.Ọnụọgụ nke mgbanwe (CV) maka nha onyonyo 28 na eserese 5b bụ 28%, ebe CV maka nha onyonyo 36 na eserese 5a gbadara ruo 9%.Nsonaazụ ndị dị n'elu na-egosi na spectrometer ọhụrụ agba agba itoolu ọ bụghị naanị na-abawanye ọnụ ọgụgụ nke agba a tụrụ n'otu oge site na asaa ruo itoolu, kamakwa ọ nwere mkpebi dị elu maka agba ọ bụla.
Ntụle nke àgwà nke ihe oyiyi kewara ekewa nke spectrometers na-emekarị na nke ọhụrụ.(a) Otu anọ nwere onyonyo kewapụrụ agba agba itoolu (C1-C9) nke spectrometer agba agba ọhụrụ agbapụtara.(b) Ihe onyonyo anọ nwere agba asaa dị iche iche (C1-C7) kpụrụ na ihe nlere anya agba asaa.Fluxes (C0) nwere ogologo ebili mmiri sitere na 400 ruo 750 nm site na isi ihe mgbapụta anọ na-agbakọta na ihe na-eme na spectrometer nke ọ bụla, n'otu n'otu.
A na-enyocha ihe ngosi nke spectrometer agba itoolu na-anwale ma gosipụta nsonaazụ nyocha na eserese 6. Rịba ama na eserese 6a na-egosi otu ihe ahụ dị ka eserese 5a, ya bụ na ogologo oge nke 4 C0 400-750 nm, a na-achọpụta ihe oyiyi 36 niile. (4 otu C1–C9).N'ụzọ megidere nke ahụ, dị ka egosiri na Fig. 6b-j, mgbe C0 ọ bụla nwere otu ogologo wavelength nke 530, 550, 570, 590, 610, 630, 650, 670, ma ọ bụ 690 nm, ọ fọrọ nke nta ka ọ bụrụ nanị ihe oyiyi anọ kwekọrọ (anọ). otu achọpụtara C1, C2, C3, C4, C5, C6, C7, C8 ma ọ bụ C9).Otú ọ dị, ụfọdụ n'ime ihe oyiyi ndị dị n'akụkụ ihe oyiyi anọ kwekọrọ na-adịghị ike n'ihi na C1-C9 mgbasa ozi nke egosiri na Fig. 4b na-adaba ntakịrị na C0 ọ bụla nwere eriri 10 nm na ogologo ogologo ogologo dị ka akọwara na usoro ahụ.Nsonaazụ ndị a dabara na ụdị mgbasa ozi C1-C9 egosiri na fig.4b na vidiyo mgbakwunye 2 na 3. N'ikwu ya n'ụzọ ọzọ, spectrometer agba agba itoolu na-arụ ọrụ dịka a tụrụ anya ya dabere na nsonaazụ egosiri na fig.4b.Ya mere, a na-ekwubi na ihe oyiyi ike nkesa C1-C9 bụ ụdị dị iche iche nke C0 ọ bụla.
Njirimara Spectral nke spectrometer agba agba itoolu.Igwe spectrometer agba agba ọhụrụ nke itoolu na-ewepụta ihe onyonyo anọ nwere agba agba itoolu (C1-C9) mgbe ọkụ ihe ahụ mere (C0 anọ) nwere ogologo ogologo (a) 400-750 nm (dị ka egosiri na Figure 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, n'otu n'otu.
A na-eji spectrometer agba itoolu emepụtara maka electrophoresis capillary anọ (maka nkọwa, lee ihe mgbakwunye)31,34,35.Matrix capillary anọ nwere capillaries anọ (dayameta mpụta 360 μm na dayameta dị n'ime 50 μm) dị na etiti 1 mm na saịtị irradiation laser.Nlereanya nwere iberibe DNA nke eji esiji 8, ya bụ FL-6C (ede 1), JOE-6C (dye 2), dR6G (ede 3), TMR-6C (ede 4), CXR-6C (ede 5), TOM- 6C (ede 6), LIZ (dye 7), na WEN (dye 8) na-arịgo n'usoro nke fluorescent wavelength, kewapụrụ na nke ọ bụla n'ime capillaries anọ (nke a na-akpọ Cap1, Cap2, Cap3, na Cap4).Igwe ọkụ na-akpata laser sitere na Cap1-Cap4 jikọtara ya na ọtụtụ lenses anọ wee dekọọ n'otu oge yana spectrometer agba itoolu.Ike ike nke agba itoolu (C1-C9) fluorescence n'oge electrophoresis, ya bụ, electrophoregram agba itoolu nke capillary ọ bụla, na-egosi na Fig. 7a.A na-enweta electrophoregram nke agba itoolu dabara na Cap1-Cap4.Dị ka akụ Cap1 gosiri na eserese 7a, ọnụ ọnụ asatọ dị na electrophoregram nke ọ bụla nwere agba itoolu na-egosi otu ikuku fluorescence sitere na Dye1-Dye8, n'otu n'otu.
Ọnụ ọgụgụ nke agba asatọ n'otu oge na-eji spectrometer nke capillary anọ nwere agba itoolu.(a) agba itoolu (C1-C9) electrophoregram nke capillary ọ bụla.Ọnụ ọnụ elu asatọ ahụ egosiri site na akụ Cap1 na-egosi mpụta nke fluorescence nke agba agba asatọ (Dye1-Dye8).Agba nke akụ ndị a kwekọrọ na agba (b) na (c).(b) Ihe nleba anya nke fluorescence nke agba agba asatọ (Dye1-Dye8) kwa capillary.c Electropherograms nke agba agba asatọ (Dye1-Dye8) kwa capillary.E ji akụ na-egosi ọnụ ọgụgụ kacha elu nke iberibe DNA akpọrọ Dye7, na-egosi ogologo isi Cap4 ha.
Nkesa ike nke C1–C9 na elu asatọ ka egosiri na fig.7b, otu.N'ihi na ma C1-C9 na Dye1-Dye8 dị n'usoro ogologo ogologo, nkesa asatọ dị na Fig. 7b na-egosi ụdị fluorescence nke Dye1-Dye8 n'usoro site n'aka ekpe gaa n'aka nri.N'ime ọmụmụ ihe a, Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, na Dye8 pụtara na magenta, violet, blue, cyan, green, yellow, orange, and red, n'otu n'otu.Rịba ama na agba nke akụ ndị dị na Fig 7a kwekọrọ na agba agba dị na Fig 7b.Igwe ọkụ ọkụ C1-C9 maka ụdịdị dị iche iche dị na eserese 7b bụ nke a na-ahazi ka nchikota ha ruru otu.E nwetara ihe nlere anya ọkụ ọkụ asatọ dabara na Cap1-Cap4.Mmadụ nwere ike ịhụ n'ụzọ doro anya na nhụsianya nke fluorescence dị n'etiti agba 1-ede 8.
Dị ka egosiri na eserese 7c, maka capillary ọ bụla, a gbanwere electrophoregram nke agba itoolu dị na Figure 7a ka ọ bụrụ electropherogram nke nwere agba asatọ site na nyocha nke ọtụtụ akụkụ dabere na ụdị fluorescence asatọ dị na Figure 7b (lee ihe mgbakwunye maka nkọwa).Ebe ọ bụ na egosipụtaghị nhụta dị iche iche nke fluorescence na eserese 7a na Figure 7c, Dye1-Dye8 nwere ike ịmata ma tụọ ya n'otu n'otu n'oge ọ bụla, ọbụlagodi ma ọ bụrụ na ọnụ ọgụgụ dị iche iche nke Dye1-Dye8 fluoresce n'otu oge.Enweghị ike ime nke a site na nchọpụta agba asaa omenala31, mana enwere ike ime ya site na nchọpụta agba itoolu emepụtara.Dị ka e gosiri site na akụ Cap1 na Fig. 7c, naanị fluorescent emission singlets Dye3 (acha anụnụ anụnụ), Dye8 (red), Dye5 (green), Dye4 (cyan), Dye2 (acha odo odo), Dye1 (magenta), na Dye6 (Yellow). ) ka a na-ahụ n'usoro oge a tụrụ anya ya.Maka mgbapụta fluorescent nke agba 7 (oroma), na mgbakwunye na otu ọnụ ọnụ elu nke akụ oroma gosipụtara, a hụrụ ọtụtụ ọnụ ọnụ ndị ọzọ.Nsonaazụ a bụ n'ihi na ihe nlele ahụ nwere ụkpụrụ nha, Dye7 kpọrọ iberibe DNA nwere ogologo isi dị iche iche.Dị ka e gosiri na Figure 7c, maka Cap4 ndị a ogologo ogologo bụ 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 na 220 isi ogologo.
Akụkụ bụ isi nke spectrometer agba itoolu, nke mepụtara site na iji matriks nke enyo dichroic nke okpukpu abụọ, bụ obere nha na nhazi dị mfe.Ebe ọ bụ na n'usoro nke decachromatic mirrors n'ime nkwụnye egosiri na fig.3c etinyere ozugbo na osisi sensọ ihe oyiyi (lee Fig. S1 na S2), spectrometer agba itoolu nwere otu akụkụ dị ka ihe nkwụnye ahụ, ntụgharị 54 × 58 × 8.5 mm.(oke).Nha a pere mpe bụ iwu abụọ ma ọ bụ atọ dị obere karịa spectrometers nke na-eji grating ma ọ bụ prisms.Tụkwasị na nke ahụ, ebe ọ bụ na a haziri spectrometer agba agba itoolu ka ọkụ na-adaba n'elu ihe mmetụta ihe oyiyi ahụ, enwere ike ịnye ohere ngwa ngwa maka spectrometer agba agba itoolu na usoro dị ka microscopes, cytometers eruba, ma ọ bụ nyocha.Capillary grating electrophoresis analyzer maka ọbụna miniaturization nke sistem.N'otu oge ahụ, nha nke enyo dichroic iri na ihe nzacha bandpass ejiri na spectrometer agba itoolu bụ naanị 10 × 1.9 × 0.5 mm ma ọ bụ 15 × 1.9 × 0.5 mm.Ya mere, ihe karịrị 100 dị otú ahụ obere enyo dichroic na bandpass nzacha, n'otu n'otu, nwere ike ebipụ si a dichroic mirror na 60 mm2 bandpass filter n'otu n'otu.Ya mere, enwere ike ịmepụta ọtụtụ enyo decachromatic na ọnụ ala dị ala.
Akụkụ ọzọ nke spectrometer agba agba itoolu bụ ọmarịcha njiri mara ya.Karịsịa, ọ na-enye ohere ịnweta ihe oyiyi dị iche iche nke snapshots, ya bụ, inweta ihe oyiyi n'otu oge na ozi dị egwu.Maka onyonyo nke ọ bụla, a na-enweta ụdịdị dị iche iche na-aga n'ihu site na 520 ruo 700 nm na mkpebi nke 20 nm.N'ikwu ya n'ụzọ ọzọ, a na-achọpụta ike ọkụ nke iteghete maka onyonyo ọ bụla, ya bụ, ụgbụ 20 nm nke itoolu na-ekewa nha ogologo ogologo site na 520 ruo 700 nm.Site n'ịgbanwe njiri mara nke enyo dichroic na ihe nzacha bandpass, enwere ike idozi ogologo ogologo ogologo nke bandị itoolu na obosara nke otu ọ bụla.Enwere ike iji nchọpụta agba itoolu ọ bụghị naanị maka nha fluorescence na eserese spectral (dị ka akọwara na akụkọ a), kamakwa maka ọtụtụ ngwa ndị ọzọ a na-ahụkarị na-eji eserese spectral.Ọ bụ ezie na ihe ngosi hyperspectral nwere ike ịchọpụta ọtụtụ narị agba, a chọpụtala na ọbụna na mbelata dị ukwuu na ọnụ ọgụgụ nke agba ndị a na-achọpụta, a pụrụ ịmata ọtụtụ ihe dị n'ọhịa nke echiche na nke ziri ezi maka ọtụtụ ngwa38,39,40.N'ihi na mkpebi gbasara ohere, mkpebi spectral, na mkpebi nke nwa oge nwere mgbanwe n'ichepụta onyonyo, ibelata ọnụ ọgụgụ agba nwere ike melite mkpebi oghere na mkpebi oge.Ọ nwekwara ike iji spectrometer dị mfe dị ka nke emepụtara n'ọmụmụ ihe a wee belata ọnụ ọgụgụ nke mgbakọ na mwepụ.
N'ime ọmụmụ ihe a, agbakọtara agba agba asatọ n'otu oge site na nkewa dị iche iche nke ihe nlere anya ọkụ ha na-ekpuchi elu dabere na nchọpụta nke agba itoolu.Enwere ike ịgbakọ ihe ruru agba ite itoolu n'otu oge, na-ebikọ ọnụ na oge na oghere.Uru pụrụ iche nke spectrometer agba agba iteghete bụ oke ọkụ ya na nnukwu oghere (1 × 7 mm).Ngosipụta enyo decane nwere oke mgbasa ozi nke 92% nke ìhè sitere na oghere na nke ọ bụla n'ime oke wavelength itoolu.Ịrụ ọrụ nke iji ọkụ ihe merenụ na nso nso a sitere na 520 ruo 700 nm fọrọ nke nta ka ọ bụrụ 100%.N'ebe dị iche iche nke ogologo wavelengths, ọ dịghị diffraction grating nwere ike inye ọrụ dị elu dị otú ahụ.Ọbụlagodi na arụmọrụ diffraction nke grating diffraction karịrị 90% n'otu ogologo wavelength, ebe ọdịiche dị n'etiti ogologo wavelength ahụ na otu ogologo wavelength na-abawanye, arụmọrụ diffraction na ogologo wavele ọzọ na-ebelata41.Enwere ike ịgbatị obosara oghere ahụ na ntụzịaka nke ụgbọ elu dị na Fig 2c site na 7 mm ruo obosara nke ihe mmetụta ihe oyiyi, dị ka n'ihe gbasara ihe mmetụta ihe oyiyi ejiri mee ihe n'ọmụmụ ihe a, site n'ịgbanwe ntakịrị ihe nhazi nke decamer.
Enwere ike iji spectrometer agba agba itoolu ọ bụghị naanị maka electrophoresis capillary, dị ka egosiri na ọmụmụ a, kamakwa maka ebumnuche ndị ọzọ.Dịka ọmụmaatụ, dị ka egosiri na foto dị n'okpuru ebe a, enwere ike itinye spectrometer agba agba itoolu na microscope fluorescence.A na-egosipụta ụgbọ elu nke nlele ahụ na ihe mmetụta oyiyi nke spectrometer agba itoolu site na ebumnuche 10x.Ebe anya anya n'etiti oghere ebumnobi na ihe mmetụta onyonyo bụ 200 mm, ebe anya anya n'etiti ihe ahụ merenụ nke spectrometer agba agba itoolu na ihe mmetụta onyonyo bụ naanị 12 mm.Ya mere, a na-egbutu ihe oyiyi ahụ ka ọ bụrụ nha nke oghere (1 × 7 mm) na ụgbọ elu nke ihe na-eme ma kee ya na ihe oyiyi agba itoolu.Nke ahụ bụ, enwere ike ịse ihe onyonyo onyonyo nke agba agba itoolu na mpaghara 0.1 × 0.7 mm n'ime ụgbọ elu nlele.Tụkwasị na nke a, ọ ga-ekwe omume ịnweta ihe oyiyi dị iche iche nke agba itoolu nke nnukwu ebe dị n'elu ụgbọ elu nlele site na nyochaa ihe nlele ahụ n'ihe gbasara ebumnobi na ntụgharị ihu na Fig 2c.
Ihe mejupụtara enyo decachromatic, ya bụ M1-M9 na BP, bụ nke Asahi Spectra Co., Ltd. mere nke ọma site na iji usoro mmiri ozuzo ọkọlọtọ.Multilayer dielectric ihe etinyere n'otu n'otu na iri quartz efere 60 × 60 mm n'ogo na 0.5 mm oké, na-emezu ihe ndị a chọrọ: M1: IA = 45 °, R ≥ 90% na 520-590 nm, Tave ≥ 90% na 610- 610 nm.700 nm, M2: IA = 45°, R ≥ 90% na 520-530 nm, Tave ≥ 90% na 550-600 nm, M3: IA = 45°, R ≥ 90% na 540-550 nm, Tave 90≥ % na 570-600 nm, M4: IA = 45°, R ≥ 90% na 560-570 nm, Tave ≥ 90% na 590-600 nm, M5: IA = 45°, R ≥ 98% na 580-600 nm , R ≥ 98% na 680-700 nm, M6: IA = 45 °, Tave ≥ 90% na 600-610 nm, R ≥ 90% na 630-700 nm, M7: IA = 45 °, R ≥ 90% 620–630 nm, Taw ≥ 90% na 650–700 nm, M8: IA = 45°, R ≥ 90% na 640–650 nm, Taw ≥ 90% na 670–700 nm, M9: IA = 45° ≥ 90% na 650-670 nm, Tave ≥ 90% na 690-700 nm, BP: IA = 0 °, T ≤ 0.01% na 505 nm, Tave ≥ 95% na 530-690 nm na 530 nm na -690 nm na T ≤ 1% na 725-750 nm, ebe IA, T, Tave, na R bụ akụkụ nke ihe omume, nnyefe, nkezi nnyefe, na ìhè na-enweghị atụ.
Ìhè na-acha ọcha (C0) nke nwere ogologo 400-750 nm sitere na isi iyi ọkụ LED (AS 3000, AS ONE CORPORATION) jikọtara ya na ihe mere n'ụzọ kwụ ọtọ na DP nke ọtụtụ enyo dichroic.A na-egosi ụdị ọkụ ọkụ na-acha ọcha na foto S3.Debe tankị acrylic (akụkụ 150 × 150 × 30 mm) ozugbo n'ihu igwe enyo decamera, na-emegide PSU.A na-awụsa anwụrụ ọkụ a na-emepụta mgbe ice kpọrọ nkụ na-emikpu n'ime mmiri n'ime tankị acrylic iji hụ iyi nke agbawa C1-C9 nke agba itoolu nke na-esite n'ụdị enyo decachromatic.
N'aka nke ọzọ, a na-agafe ọkụ ọkụ collimated (C0) site na nzacha tupu ịbanye na DP.Ihe nzacha ndị ahụ bụ nzacha njupụta na-anọpụ iche na njupụta anya nke 0.6.Mgbe ahụ jiri nzacha moto (FW212C, FW212C, Thorlabs).N'ikpeazụ, gbanye ND nzacha azụ.Ihe nzacha bandwit nke nzacha bandpass itoolu dabara na C9, C8, C7, C6, C5, C4, C3, C2 na C1, n'otu n'otu.A na-etinye cell quartz nwere akụkụ dị n'ime nke 40 (ogologo anya) x 42.5 (ịdị elu) x 10 mm (obosara) n'ihu ọtụtụ enyo decochromatic, na-emegide BP.A na-enyezi anwụrụ ọkụ ahụ site na tube n'ime cell quartz iji nọgide na-enwe mmetụta nke anwụrụ ọkụ na cell quartz iji hụ anya mmiri C1-C9 nwere agba itoolu nke na-esite na nhazi enyo decachromatic.
Ededere vidiyo nke iyi ọkụ gbawara agba agba itoolu nke na-esite n'ọtụtụ enyo decanic na ọnọdụ oge na-adịghị na iPhone XS.Were foto nke ihe nkiri ahụ na 1 fps wee chịkọta onyonyo iji mepụta vidiyo na 30fps (maka vidiyo nhọrọ 1) ma ọ bụ 24fps (maka vidiyo nhọrọ 2 na 3).
Tinye efere igwe anaghị agba nchara 50µm (nwere oghere dayameta 50 µm anọ na etiti 1 mm) na efere mgbasa.A na-agbanye ọkụ nke nwere ogologo 400-750 nm n'elu efere diffuser, nke enwetara site na ịfefe ọkụ site na oriọna halogen site na nzacha nnyefe dị mkpirikpi nke nwere nkwụsị nke 700 nm.A na-egosi ụdị ọkụ ahụ na foto mgbakwunye S4.N'aka nke ọzọ, ọkụ na-agafe n'otu n'ime ihe nzacha bandpass 10 nm gbadoro ụkwụ na 530, 550, 570, 590, 610, 630, 650, 670 na 690 nm wee daa efere diffuser.N'ihi nke a, e hiwere isi ihe radieshon anọ nwere dayameta nke φ50 μm na ogologo wavelength dị iche iche na efere igwe anaghị agba nchara chere efere mgbasa ozi.
A na-etinye eriri capillary anọ nwere lenses anọ na spectrometer agba itoolu dị ka egosiri na foto 1 na 2. C1 na C2.Capillaries anọ na lenses anọ bụ otu ihe na ọmụmụ gara aga31,34.A na-agbanye ọkụ laser nke nwere ogologo 505 nm na ike nke 15 mW n'otu oge na ọbụna site n'akụkụ ruo n'isi ihe mgbapụta nke capillaries anọ.A na-ejikọta fluorescence nke ebe ọ bụla na-ekupụta site na oghere kwekọrọ wee kewaa n'ime iyi agba agba itoolu site n'ụdị enyo decachromatic.Agbanyezie iyi iyi 36 ndị ahụ ozugbo n'ime ihe mmetụta onyonyo CMOS (C11440-52U, Hamamatsu Photonics K·K.), na edekọ onyonyo ha n'otu oge.
ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Applied Biosystems), 4 µl GeneScan™ 600 LIZ™ dye agwakọtara maka capillary ọ bụla site na ịgwakọta 1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), 1µl ọkọlọtọ nha nha nha.v2.0 (Thermo Fisher Scientific) na 14 µl mmiri.Standardkpụrụ Matrix PowerPlex® 6C nwere iberibe DNA isii akpọrọ ya na agba isii: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, na WEN, n'usoro ogologo ikuku kacha.A naghị ekpughe ogologo isi nke iberibe DNA ndị a, mana a maara usoro ogologo ogologo ogologo nke DNA nke akpọrọ WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C na TOM-6C.Ngwakọta a dị na ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit nwere iberibe DNA nke akpọrọ dR6G agba.A naghị egosikwa ogologo ntọala nke iberibe DNA.GeneScan™ 600 LIZ™ Dye Size Standard v2.0 gụnyere iberibe DNA nwere akara 36 LIZ.Ogologo mpempe akwụkwọ ndị a dị afọ iri abụọ, 40, 100 60, mmadụ iri abụọ na ise, mmadụ iri abụọ na ise, 260, mmadụ iri abụọ na ise, 260, 300, 314, 300, 340, 340, 340, 340, 340, 340, 340, 340, 340, 340, 340, 340, 340, 340, 340, 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 na 600 isi.A na-ewepụ ihe nlele ahụ na 94 Celsius C maka nkeji 3, wee mee ka ọ dị jụụ na ice maka nkeji 5.A na-etinye ihe nlele n'ime capillary ọ bụla na 26 V / cm maka 9 s wee kewapụ ya na capillary ọ bụla jupụtara na ngwọta POP-7 ™ polymer (Thermo Fisher Scientific) na ogologo dị irè nke 36 cm na voltaji nke 181 V / cm na otu. akụkụ nke 60°.SITE.
Ihe niile enwetara ma ọ bụ nyochaa n'oge ọmụmụ ihe a gụnyere n'ime akụkọ a na-ebipụta na ozi ndị ọzọ.Data ndị ọzọ metụtara ọmụmụ ihe a dị site n'aka ndị odee dị iche iche na arịrịọ ezi uche dị na ya.
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