insimbi engagqwali 321 8 * 1.2 tube coiled for heat exchanger

Amashubhu eCapillary

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Kwakhiwe i-spectrometer ehlangene kakhulu (54 × 58 × 8.5 mm) kanye nendawo ebanzi (1 × 7 mm) enemibala eyisishiyagalolunye, "ehlukaniswe kabili" ngoxhaxha lwezibuko eziyishumi ze-dichroic, ezasetshenziswa ekuthwebuleni izithombe ngokushesha.Isigameko sokugeleza kokukhanya esinesigaba esiphambanayo esincane kunosayizi wokuvula ihlukaniswe yaba umucu oqhubekayo ongu-20 nm ububanzi nobubanzi obuyisishiyagalolunye obunamaza amaphakathi angu-530, 550, 570, 590, 610, 630, 650, 670 no-690 nm.Izithombe zemibala egeleza eyisishiyagalolunye zilinganiswa ngesikhathi esifanayo nomuzwa wesithombe.Ngokungafani nezinhlaka ezijwayelekile zesibuko se-dichroic, isibuko se-dichroic esithuthukisiwe sinokucushwa okuyingqayizivele kweziqephu ezimbili, okungagcini nje ngokukhuphula inani lemibala engalinganiswa ngesikhathi esisodwa, kodwa futhi kuthuthukisa ukucaca kwesithombe kokusakaza kombala ngakunye.I-spectrometer ethuthukisiwe enemibala eyisishiyagalolunye isetshenziselwa i-electrophoresis engama-capillary amane.Ukuhlaziywa kobuningi ngesikhathi esisodwa kodayi abayisishiyagalombili abafudukayo ngesikhathi esisodwa ku-capillary ngayinye kusetshenziswa i-laser-induced fluorescence enemibala eyisishiyagalolunye.Njengoba i-spectrometer enemibala eyisishiyagalolunye ingeyona nje kuphela i-ultra-encane futhi engabizi, kodwa futhi ine-flux ephezulu ekhanyayo kanye nokulungiswa kwe-spectral okwanele kwezinhlelo zokusebenza eziningi ze-imaging, ingasetshenziswa kabanzi emikhakheni ehlukahlukene.
I-hyperspectral ne-multispectral imaging isiyingxenye ebalulekile ye-astronomy2, i-remote sensing for Earth observation3,4, ukulawulwa kwekhwalithi yokudla namanzi5,6, ukongiwa kobuciko kanye ne-archeology7, forensics8, surgery9, biomedical analysis and diagnostics10,11 njll. Inkambu 1 Ubuchwepheshe obubalulekile ,12,13.Izindlela zokulinganisa i-spectrum yokukhanya okukhishwa iphuzu ngalinye eliphumayo emkhakheni wokubuka zihlukaniswe (1) ukuskena kwamaphuzu (“umshanelo”)14,15, (2) ukuskena ngomugqa (“panicle”)16,17,18 , (3) ubude buskena amagagasi19,20,21 kanye (4) nezithombe22,23,24,25.Endabeni yazo zonke lezi zindlela, ukulungiswa kwendawo, ukulungiswa kwe-spectral kanye nokulungiswa kwesikhashana kunobudlelwane bokuhwebelana9,10,12,26.Ukwengeza, ukuphuma kokukhanya kunomthelela obalulekile ekuzweleni, okungukuthi isilinganiso sesignali-kuya-nomsindo kumfanekiso we-spectral26.I-flux ekhanyayo, okungukuthi, ukusebenza kahle kokusebenzisa ukukhanya, ilingana ngokuqondile nesilinganiso senani langempela elilinganiselwe lokukhanya lephoyinti ngalinye elikhanyayo ngesikhathi seyunithi ukuya enanini eliphelele lokukhanya lobubanzi obulinganisiwe bobude beza.Isigaba (4) siyindlela efanelekile lapho amandla noma i-spectrum yokukhanya okukhishwa iphuzu ngalinye elikhiphayo kushintsha ngesikhathi noma lapho indawo yephoyinti elikhiphayo ngalinye ishintsha ngokuhamba kwesikhathi ngenxa yokuthi i-spectrum yokukhanya okukhishwa yiwo wonke amaphuzu aphumayo ikalwa ngesikhathi esisodwa.24.
Iningi lalezi zindlela ezingenhla zihlanganiswe nama-spectrometer amakhulu, ayinkimbinkimbi kanye/noma abizayo kusetshenziswa ama-grating angu-18 noma ama-prism angu-14, 16, 22, 23 amakilasi (1), (2) kanye (4) noma 20, 21 ama-filter disks, izihlungi eziwuketshezi. .Izihlungi ezi-Crystalline eziguqulekayo (LCTF)25 noma izihlungi ezi-acousto-optic tunable (AOTF)19 zesigaba (3).Ngokuphambene, ama-spectrometer esigaba (4) anezibuko eziningi mancane futhi awabizi ngenxa yokucushwa kwawo okulula27,28,29,30.Ngaphezu kwalokho, banokushintshashintsha okukhanyayo okuphezulu ngenxa yokuthi ukukhanya okwabelwana ngesibuko ngasinye se-dichroic (okungukuthi, ukukhanya okudlulisiwe nokuboniswayo kokukhanya kwesigameko esibukweni ngasinye se-dichroic) kusetshenziswa ngokugcwele nangokuqhubekayo.Nokho, inani lamabhendi e-wavelength (okungukuthi imibala) okufanele likalwe ngesikhathi esisodwa likhawulelwe cishe kumane.
Ukuthwebula kwe-Spectral okusekelwe ekutholweni kwe-fluorescence kuvame ukusetshenziselwa ukuhlaziywa kwe-multiplex ekutholweni kwe-biomedical kanye nokuxilongwa 10, 13.Ekuphindaphindeni, njengoba ama-analyte amaningi (isb, i-DNA ethize noma amaprotheni) alebulwe ngodayi abahlukene be-fluorescent, i-analyte ngayinye ekhona endaweni ngayinye ekhishwayo emkhakheni wokubuka ilinganiswa kusetshenziswa ukuhlaziywa kwezingxenye eziningi.32 yephula i-spectrum ye-fluorescence etholiwe ekhishwa yiphuzu ngalinye eliphumayo.Phakathi nale nqubo, odayi abahlukene, ngamunye okhipha i-fluorescence ehlukile, bangahlangana, okungukuthi, bahlale endaweni kanye nesikhathi.Njengamanje, inani eliphezulu lodayi elingajatshuliswa i-laser beam eyodwa liyisishiyagalombili33.Lo mkhawulo ongaphezulu awunqunywa ukulungiswa kwe-spectral (okungukuthi, inani lemibala), kodwa ububanzi be-spectrum ye-fluorescence (≥50 nm) kanye nenani lokushintsha kodayi be-Stokes (≤200 nm) ku-FRET (kusetshenziswa i-FRET)10 .Kodwa-ke, inani lemibala kufanele libe likhulu noma lilingane nenani lamadayi ukuze kuqedwe ukunqwabelana kodayi abaxubile31,32.Ngakho-ke, kuyadingeka ukwandisa inani lemibala elinganisiwe kanyekanye ibe yisishiyagalombili noma ngaphezulu.
Muva nje, i-ultra-compact heptachroic spectrometer (esebenzisa inqwaba yezibuko ze-heptychroic kanye nenzwa yesithombe ukukala ama-fluorescent fluxes amane) yenziwe.I-spectrometer ingama-oda amabili kuya kwamathathu obukhulu obuncane kunama-spectrometer avamile kusetshenziswa ama-gratings noma ama-prisms34,35.Nokho, kunzima ukubeka izibuko ezingaphezu kweziyisikhombisa ze-dichroic ku-spectrometer futhi ngesikhathi esisodwa ukala imibala engaphezu kweyisikhombisa36,37.Ngokukhula kwenani lezibuko ze-dichroic, umehluko omkhulu ebude bezindlela zokubonwa kwe-dichroic light fluxes uyakhula, futhi kuba nzima ukubonisa konke ukukhanya endizeni eyodwa yezinzwa.Ubude bendlela ye-optical ende kunazo zonke ye-flux yokukhanya nayo iyakhuphuka, ngakho ububanzi bendawo yokuvula i-spectrometer (okungukuthi ububanzi bokukhanya obuningi obuhlaziywa i-spectrometer) buyancipha.
Ukusabela ezinkingeni ezingenhla, i-spectrometer ye-ultra-compact eyisishiyagalolunye enemibala eyisishiyagalolunye enezingqimba ezimbili zesibuko se-decachromatic "dichroic" kanye nenzwa yesithombe yokuthwebula kwesithombe esisheshayo [isigaba (4)].Uma kuqhathaniswa nama-spectrometer angaphambilini, i-spectrometer ethuthukisiwe inomehluko omncane kubude bendlela ye-optical esiphezulu kanye nobude obuncane bendlela yokubona.Isetshenziswe ku-four-capillary electrophoresis ukuze kutholwe i-laser-induced eyisishiyagalolunye-color fluorescence kanye nokubala ukufuduka ngesikhathi esisodwa kodayi abayisishiyagalombili ku-capillary ngayinye.Njengoba i-spectrometer ethuthukisiwe ingeyona nje kuphela i-ultra-encane futhi engabizi, kodwa futhi ine-flux ephezulu ekhanyayo kanye nesixazululo esanele se-spectral yezinhlelo zokusebenza eziningi ze-imaging, ingasetshenziswa kabanzi emikhakheni ehlukahlukene.
I-spectrometer yendabuko enemibala eyisishiyagalolunye ikhonjiswe emkhiwaneni.1a.Idizayini yayo ilandela eye-spectrometer yangaphambilini encane kakhulu enemibala eyisikhombisa engu-31. Iqukethe izibuko eziyisishiyagalolunye ze-dichroic ezihlelwe ngokuvundlile nge-engeli engu-45° kwesokudla, futhi inzwa yesithombe (S) itholakala ngaphezu kwezibuko eziyisishiyagalolunye ze-dichroic.Ukukhanya okungena kusuka ngezansi (C0) kuhlukaniswe ngezibuko eziyisishiyagalolunye ze-dichroic zibe ukugeleza kokukhanya okuyisishiyagalolunye okukhuphukayo (C1, C2, C3, C4, C5, C6, C7, C8 and C9).Yonke imibala eyisishiyagalolunye inikezwa inzwa yesithombe futhi itholwa kanyekanye.Kulolu cwaningo, i-C1, i-C2, i-C3, i-C4, i-C5, i-C6, i-C7, i-C8, ne-C9 ilandelana nge-wavelength futhi imelelwa i-magenta, i-violet, eluhlaza okwesibhakabhaka, e-cyan, eluhlaza, ephuzi, ewolintshi, ebomvu-wolintshi, kanye obomvu, ngokulandelana.Nakuba lezi zimpawu zemibala zisetshenziswa kulo mbhalo, njengoba kuboniswe kuMfanekiso 3, ngoba zihlukile nemibala yangempela ebonwa iso lomuntu.
Imidwebo eyisikimu yama-spectrometer ajwayelekile kanye namasha anemibala eyisishiyagalolunye.(a) I-spectrometer evamile enemibala eyisishiyagalolunye enoxhaxha lwezibuko eziyisishiyagalolunye ze-dichroic.(b) I-spectrometer entsha yemibala eyisishiyagalolunye enezingqimba ezimbili zesibuko se-dichroic.Isigameko esishintshashintshayo sokukhanya kwe-C0 sihlukaniswe ngama-fluxes okukhanya anemibala ayisishiyagalolunye C1-C9 futhi atholwa inzwa yesithombe u-S.
I-spectrometer entsha ethuthukisiwe enemibala eyisishiyagalolunye ine-grating yesibuko se-dichroic enezingqimba ezimbili kanye nenzwa yesithombe, njengoba kuboniswe ku-Fig. 1b.Esigabeni esingezansi, izibuko ezinhlanu ze-dichroic zitshekele ngo-45° kwesokudla, ziqondaniswe kwesokudla ukusuka maphakathi nohlu lwama-decamers.Ezingeni eliphezulu, izibuko ezinhlanu ezengeziwe ze-dichroic zitshekiswa ngo-45° kwesokunxele futhi zitholakala kusukela phakathi nendawo ukuya kwesokunxele.Isibuko se-dichroic kwesokunxele sesendlalelo esingezansi kanye nesibuko se-dichroic kwesokudla sesendlalelo esingaphezulu kuyagqagqana.Isigameko sokukhanya kokukhanya (C0) sihlukaniswa kusukela ngezansi sibe ama-chromatic fluxes amane aphumayo (C1-C4) ngezibuko ezinhlanu ze-dichroic kwesokudla kanye nama-chromatic fluxes amahlanu aphumayo (C5-C4) ngezibuko ezinhlanu ze-dichroic kwesokunxele C9).Njengama-spectrometer avamile anemibala eyisishiyagalolunye, yonke imifudlana yemibala eyisishiyagalolunye ijovwa ngokuqondile kunzwa yesithombe (S) futhi itholwe kanyekanye.Ukuqhathanisa Izibalo 1a kanye no-1b, umuntu angabona ukuthi esimweni se-spectrometer entsha yemibala eyisishiyagalolunye, kokubili umehluko omkhulu kanye nobude bendlela yokubona ende yemibala eyisishiyagalolunye eguquguqukayo ihhafu.
Ukwakhiwa okunemininingwane ye-ultra-small yohlelo lwesibuko se-dichroic enezendlalelo ezimbili eziwu-29 mm (ububanzi) × 31 mm (ukujula) × 6 mm (ubude) kuboniswe kuMfanekiso 2. Uhlu lwesibuko sedesimali se-dichroic luqukethe izibuko ezinhlanu ze-dichroic kwesokudla. (M1-M5) kanye nezibuko ezinhlanu ze-dichroic kwesokunxele (M6-M9 nenye i-M5), isibuko ngasinye se-dichroic sigxilwe kubakaki be-aluminium ephezulu.Zonke izibuko ze-dichroic ziyanyakaziswa ukuze kunxephezele ukususwa endaweni efanayo ngenxa yokunqamuka kokugeleza kwezibuko.Ngezansi kwe-M1, isihlungi se-band-pass (BP) silungisiwe.Ubukhulu be-M1 ne-BP bungu-10mm (uhlangothi olude) x 1.9mm (uhlangothi olufushane) x 0.5mm (ubukhulu).Ubukhulu bezibuko ze-dichroic ezisele ziyi-15 mm × 1.9 mm × 0.5 mm.I-matrix pitch phakathi kwe-M1 ne-M2 ingu-1.7 mm, kuyilapho i-matrix pitch yezinye izibuko ze-dichroic ingu-1.6 mm.Emkhiwaneni.I-2c ihlanganisa ukukhanya kokukhanya kwesigameko C0 kanye nokushintshashintsha kokukhanya okunemibala eyisishiyagalolunye C1-C9, kuhlukaniswe i-matrix ye-de-chamber yezibuko.
Ukwakhiwa kwe-two-layer dichroic mirror matrix.(a) Umbono wokubuka kanye (b) nokubuka okunqamulayo kohlu lwesibuko se-dichroic enezendlalelo ezimbili (ubukhulu 29 mm x 31 mm x 6 mm).Iqukethe izibuko ezinhlanu ze-dichroic (M1-M5) ezitholakala kungqimba olungezansi, izibuko ezinhlanu ze-dichroic (M6-M9 nenye i-M5) etholakala kungqimba olungaphezulu, kanye nesihlungi se-bandpass (BP) esingaphansi kwe-M1.(c) Ukubuka kwesigaba esiphambanayo kuhlangothi oluqondile, olunokugqagqana kwe-C0 ne-C1-C9.
Ububanzi bembobo endaweni evundlile, ekhonjiswe ububanzi C0 ku-Fig. 2, c, buyi-1 mm, futhi ohlangothini lwe-perpendicular endizeni ye-Fig. 2, c, enikezwe ukwakheka kwebakaki le-aluminium, - 7 mm.Okusho ukuthi, i-spectrometer entsha yemibala eyisishiyagalolunye inosayizi omkhulu wokuvula we-1 mm × 7 mm.Indlela ye-optical ye-C4 yinde kunazo zonke phakathi kwe-C1-C9, futhi indlela ye-optical ye-C4 ngaphakathi kwe-dichroic mirror array, ngenxa yobukhulu obungenhla obuncane (29 mm × 31 mm × 6 mm), ingu-12 mm.Ngesikhathi esifanayo, ubude bendlela ye-optical ye-C5 bufushane kakhulu phakathi kwe-C1-C9, futhi ubude bendlela ye-C5 bungu-5.7mm.Ngakho-ke, umehluko omkhulu kubude bendlela ye-optical ngu-6.3 mm.Ubude bendlela yokubona engenhla bulungiswa kubude bendlela yokubona yokudlulisa okubonakalayo kwe-M1-M9 ne-BP (kusuka ku-quartz).
Izakhiwo ze-spectral ze-М1−М9 ne-VR zibalwa ukuze ukuguquguquka okungu-С1, С2, С3, С4, С5, С6, С7, С8 kanye no-С9 kububanzi be-wavelength 520–540, 540–560, 560–8580, 560–8580, 580, 580 -600 , 600-620, 620-640, 640-660, 660-680, kanye no-680-700 nm, ngokulandelana.
Isithombe se-matrix ekhiqiziwe yezibuko ze-decachromatic sikhonjiswe ku-Fig. 3a.I-M1-M9 ne-BP zinamathiselwe ku-45° ukuthambeka kanye nendiza evundlile yosekelo lwe-aluminium, ngokulandelana, kuyilapho i-M1 ne-BP zifihliwe ngemuva kwesibalo.
Ukukhiqizwa kohlu lwezibuko ze-decan kanye nokuboniswa kwazo.(a) Uxhaxha lwezibuko ze-decachromatic ezakhiwe.(b) Isithombe esihlukanisayo esingu-1 mm × 7 mm esinemibala eyisishiyagalolunye esivezwa eshidini lephepha elibekwe ngaphambi kohlu lwezibuko ze-decachromatic futhi elikhanyiswe ngemuva ngokukhanya okumhlophe.(c) Uxhaxha lwezibuko ze-decochromatic ezikhanyiswe ngokukhanya okumhlophe ukusuka ngemuva.(d) Umfudlana ohlukanisayo onemibala eyisishiyagalolunye ophuma kuhlelo lwesibuko se-decane, obonwa ngokubeka i-acrylic canister egcwele intuthu ngaphambi kwesibuko se-decane ku-c futhi kube mnyama igumbi.
I-spectra yokudlulisa elinganisiwe ye-M1-M9 C0 nge-engeli yesigameko engu-45° kanye ne-spectrum yokudlulisela elinganisiwe ye-BP C0 nge-engeli yesigameko engu-0 ° iboniswa kumakhiwane.4a.I-spectra yokudlulisela ye-C1-C9 ehlobene ne-C0 iboniswa ku-Fig.4b.Lezi zibukeli zibalwe kusukela ku-spectra ku-Fig.4a ngokuhambisana nendlela ye-optical C1-C9 ku-Fig. 4a.1b no2c.Isibonelo, i-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)], lapho TS(X) kanye [ 1 − TS(X)] iwukudluliswa nokubukeka kwe-spectra ka-X, ngokulandelanayo.Njengoba kuboniswe kuMfanekiso 4b, umkhawulokudonsa (umkhawulokudonsa ≥50%) we-C1, C2, C3, C4, C5, C6, C7, C8 kanye ne-C9 ngu-521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 kanye no-682-699 nm.Le miphumela ihambisana nobubanzi obuthuthukisiwe.Ngaphezu kwalokho, ukusebenza kahle kokukhanya kwe-C0 kuphezulu, okungukuthi, isilinganiso esiphezulu sokudlulisa ukukhanya kwe-C1-C9 singama-92%.
I-spectra yokudlulisa yesibuko se-dichroic kanye nokuhlukaniswa kwemibala eyisishiyagalolunye.(a) I-spectra yokudlulisela elinganiselwe ye-M1-M9 ku-45° izehlakalo kanye ne-BP ku-0° izehlakalo.(b) I-spectra yokudlulisela ye-C1–C9 ehlobene ne-C0 ebalwa kusukela ku-(a).
Emkhiwaneni.I-3c, izibuko ze-dichroic eziningi zitholakala ziqondile, ukuze uhlangothi lwayo lwesokudla ku-Fig. 3a luhlangothini olungaphezulu futhi i-beam emhlophe ye-LED ehlanganisiwe (C0) ikhanyiselwe emuva.Uhlu lwezibuko ze-decachromatic eziboniswe ku-Figure 3a zifakwe ku-adaptha engu-54 mm (ubude) × 58 mm (ukujula) × 8.5 mm (ubukhulu).Emkhiwaneni.3d, ngaphezu kwesimo esiboniswe kumfanekiso.3c, ithangi le-acrylic eligcwele intuthu labekwa phambi kwezibuko ze-decochromatic, izibani zegumbi zivaliwe.Ngenxa yalokho, kuyabonakala imifudlana eyisishiyagalolunye ye-dichroic ethangini, ephuma kuxhaxha lwezibuko ze-decachromatic.Ukusakaza ngakunye okuhlukanisayo kunesigaba esiphambanayo esingunxande esinobukhulu obungu-1 × 7 mm, obuhambisana nosayizi wokuvula we-spectrometer entsha yemibala eyisishiyagalolunye.Emfanekisweni 3b, ishidi lephepha libekwa phambi koxhaxha lwezibuko ze-dichroic ku-Figure 3c, futhi isithombe esingu-1 x 7 mm semifudlana eyisishiyagalolunye ye-dichroic evezwa ephepheni ibonwa ukusuka lapho kuhamba khona iphepha.imifudlana.Imibala eyisishagalolunye igeleza kufig.3b kanye no-d yi-C4, C3, C2, C1, C5, C6, C7, C8 kanye no-C9 ukusuka phezulu kuye phansi, okungabuye kubonakale kuzibalo 1 no-2. 1b kanye no-2c.Zibonwa ngemibala ehambisana namaza azo.Ngenxa yokuqina kokukhanya okumhlophe okuphansi kwe-LED (bona i-Supplementary Fig. S3) kanye nokuzwela kwekhamera yombala esetshenziselwa ukuthwebula i-C9 (682–699 nm) ku-Fig. Okunye ukugeleza okuhlukanisayo akunamandla.Ngokufanayo, i-C9 ibibonakala kancane ngeso lenyama.Khonamanjalo, i-C2 (umfudlana wesibili osuka phezulu) ibukeka iluhlaza kuMfanekiso 3, kodwa ibukeka iphuzi kakhulu ngeso lenyama.
Ushintsho olusuka kuMfanekiso 3c ukuya ku-d luboniswa ku-Supplementary Video 1. Ngokushesha ngemva kokuba ukukhanya okumhlophe okuvela ku-LED kudlule kuhlelo lwesibuko se-decachromatic, kuhlukana ngesikhathi esisodwa kube imifudlana eyimibala eyisishiyagalolunye.Ekugcineni, intuthu ku-vat yashabalala kancane kancane ukusuka phezulu kuya phansi, kangangokuthi izimpushana ezinemibala eyisishiyagalolunye nazo zanyamalala ukusuka phezulu kuye phansi.Ngokuphambene, ku-Supplementary Video 2, lapho ubude begagasi besigameko sokugeleza kokukhanya ochungechungeni lwezibuko ze-decachromatic bushintshiwe ukusuka kubude ukuya kwabafushane ngokulandelana kuka-690, 671, 650, 632, 610, 589, 568, 550 kanye no-532 nm. ., Kuboniswa kuphela ukusakaza okuhlukanisayo okuhambisanayo kokuhlukaniswa kwemifudlana eyisishiyagalolunye ngokulandelana kwe-C9, C8, C7, C6, C5, C4, C3, C2, ne-C1.Indawo ye-acrylic reservoir ithathelwa indawo yi-quartz pool, futhi ama-flakes of shunted flow ngayinye angabonwa ngokucacile ukusuka endaweni eyehlelayo eya phezulu.Ngaphezu kwalokho, ividiyo engaphansi 3 ihlelwa ngendlela yokuthi ingxenye yoshintsho lwamaza evidiyo engaphansi 2 iphinde idlalwe.Lokhu ukuvezwa okucacile kwezici zezibuko ze-decochromatic.
Imiphumela engenhla ibonisa ukuthi i-decachromatic mirror array ekhiqiziwe noma i-spectrometer entsha yemibala eyisishiyagalolunye isebenza ngendlela ehlosiwe.I-spectrometer entsha enemibala eyisishiyagalolunye yakhiwe ngokukhweza izibuko ze-decachromatic ezinama-adaptha ngqo ebhodini lenzwa yesithombe.
I-flux ekhanyayo enobubanzi be-wavelength kusuka ku-400 kuya ku-750 nm, ekhishwa ngamaphuzu amane emisebe φ50 μm, etholakala ngezikhathi ze-1 mm ekuqondeni kwendiza ye-Fig. 2c, ngokulandelanayo Ucwaningo 31, 34. Uhlu lwamalensi amane luhlanganisa amalensi amane φ1 mm anobude bokugxila obungu-1.4 mm kanye ne-pitch engu-1 mm.Imifudlana enqwabelene emine (C0 emine) iyisigameko ku-DP ye-spectrometer entsha enemibala eyisishiyagalolunye, ehlukaniswe ngezikhawu ezingu-1 mm.Uhlu lwezibuko ze-dichroic luhlukanisa ukusakaza ngakunye (C0) kube imifudlana yemibala eyisishiyagalolunye (C1-C9).Umphumela wokusakaza ongu-36 (amasethi amane e-C1-C9) abe esejovwa ngokuqondile kunzwa yesithombe ye-CMOS (S) exhunywe ngokuqondile kuhlu lwezibuko ze-dichroic.Ngenxa yalokho, njengoba kuboniswe ku-Fig. 5a, ngenxa yomehluko omncane wendlela yokubona ephezulu kanye nendlela emfushane ye-optical esiphezulu, izithombe zakho zonke ezingu-36 zokusakaza zitholwe kanyekanye futhi ngokucacile ngosayizi ofanayo.Ngokuvumelana ne-spectra engezansi (bona Umfanekiso Owengeziwe S4), ukushuba kwesithombe samaqembu amane C1, C2 kanye ne-C3 kuphansi kakhulu.Izithombe ezingamashumi amathathu nesithupha bezingu-0.57 ± 0.05 mm ngosayizi (okusho ukuthi ± SD).Ngakho-ke, ukukhuliswa kwesithombe kwaba nesilinganiso esingu-11.4.Isikhala esiqondile phakathi kwezithombe singu-1 mm (isikhala esifanayo ne-lens yamalungu afanayo) kanye nesikhala esivundlile singu-1.6 mm (isikhala esifanayo ne-dichroic mirror array).Ngenxa yokuthi usayizi wesithombe mncane kakhulu kunebanga phakathi kwezithombe, isithombe ngasinye singalinganiswa ngokuzimela (nge-crosstalk ephansi).Phakathi naleso sikhathi, izithombe zemifudlana engamashumi amabili nesishiyagalombili ezirekhodwe i-spectrometer evamile yemibala eyisikhombisa esetshenziswe ocwaningweni lwethu lwangaphambilini ziboniswa ku-Fig. 5 B. Uhlu lwezibuko eziyisikhombisa ze-dichroic lwakhiwe ngokususa izibuko ezimbili ezingakwesokudla kusuka kuhlu lwe-dichroic eyisishiyagalolunye. izibuko kuMfanekiso 1a.Akuzona zonke izithombe ezibukhali, usayizi wesithombe uyakhuphuka usuka ku-C1 uye ku-C7.Izithombe ezingamashumi amabili nesishiyagalombili zingu-0.70 ± 0.19 mm ngosayizi.Ngakho-ke, kunzima ukugcina ukulungiswa okuphezulu kuzo zonke izithombe.I-coefficient of variation (CV) yosayizi wesithombe 28 kuMfanekiso 5b ibingu-28%, kuyilapho i-CV yosayizi wesithombe 36 kuFigure 5a yehle yaba ngu-9%.Imiphumela engenhla ibonisa ukuthi i-spectrometer entsha enemibala eyisishiyagalolunye ayikhulisi nje kuphela inani lemibala elinganiswa ngasikhathi sinye isuka kweyisikhombisa iye kweyisishiyagalolunye, kodwa futhi inokulungiswa kwesithombe okuphezulu kumbala ngamunye.
Ukuqhathaniswa kwekhwalithi yesithombe esihlukanisiwe esakhiwe ama-spectrometer avamile namasha.(a) Amaqembu amane ezithombe ezihlukene ezinemibala eyisishiyagalolunye (C1-C9) akhiqizwe i-spectrometer entsha yemibala eyisishiyagalolunye.(b) Amasethi amane ezithombe ezihlukene ezinemibala eyisikhombisa (C1-C7) ezakhiwe nge-spectrometer evamile enemibala eyisikhombisa.Ama-Fluxes (C0) anobude obungamaza ukusuka ku-400 kuye ku-750 nm ukusuka ezindaweni ezine ezikhishwayo ayahlanganiswa futhi enzeka ku-spectrometer ngayinye, ngokulandelana.
Izici ze-spectral ze-spectrometer yemibala eyisishiyagalolunye zahlolwa ngokuhlolwa futhi imiphumela yokuhlola iboniswa kuMfanekiso 6. Qaphela ukuthi Umfanekiso 6a ubonisa imiphumela efanayo njengoMfanekiso 5a, okungukuthi kumaza wamaza angu-4 C0 400–750 nm, zonke izithombe ezingu-36 zitholwa. (4 amaqembu C1–C9).Ngokuphambene, njengoba kuboniswe ku-Fig. 6b–j, lapho i-C0 ngayinye inobude obuthile obungu-530, 550, 570, 590, 610, 630, 650, 670, noma 690 nm, cishe kunezithombe ezine kuphela ezihambisanayo (ezine amaqembu athole u-C1, C2, C3, C4, C5, C6, C7, C8 noma C9).Kodwa-ke, ezinye zezithombe ezincikene nezithombe ezine ezihambisanayo zitholwa ngobuthaka kakhulu ngenxa yokuthi i-spectra yokudlulisa ye-C1–C9 eboniswe ku-Fig. 4b idlula kancane futhi i-C0 ngayinye inebhendi engu-10 nm kubude begagasi obuthile njengoba kuchazwe endleleni.Le miphumela ihambisana ne-C1-C9 transmission spectra eboniswe ku-Fig.4b kanye namavidiyo engeziwe 2 no-3. Ngamanye amazwi, i-spectrometer yombala eyisishiyagalolunye isebenza njengoba kulindelekile ngokusekelwe emiphumeleni ekhonjiswe kumfanekiso.4b.Ngakho-ke, kuphethwa ngokuthi ukusabalalisa kokuqina kwesithombe i-C1-C9 yi-spectrum ye-C0 ngayinye.
Izici ze-Spectral ze-spectrometer enemibala eyisishiyagalolunye.I-spectrometer entsha enemibala eyisishiyagalolunye ikhiqiza amasethi amane ezithombe ezihlukene ezinemibala eyisishiyagalolunye (C1-C9) lapho ukukhanya kwesigameko (C0) kunobude begagasi obungu-(a) 400-750 nm (njengoba kuboniswe kuMfanekiso 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, ngokulandelana.
I-spectrometer ethuthukisiwe enemibala eyisishiyagalolunye yasetshenziselwa i-electrophoresis engama-capillary amane (ngemininingwane, bheka Izinto Ezingeziwe)31,34,35.I-four-capillary matrix iqukethe ama-capillary amane (ububanzi obungaphandle obungu-360 μm kanye nobubanzi bangaphakathi bungu-50 μm) atholakala ngezikhawu ezingu-1 mm endaweni yokuthela ngemisebe ye-laser.Amasampula aqukethe izingcezu ze-DNA ezilebulwe ngodayi abangu-8, okungu-FL-6C (udayi 1), JOE-6C (udayi 2), dR6G (udayi 3), TMR-6C (udayi 4), CXR-6C (udayi 5), TOM- 6C (udayi 6), LIZ (udayi 7), kanye ne-WEN (udayi 8) ngokulandelana okunyukayo kwe-wavelength ye-fluorescent, ehlukaniswe ngama-capillaries amane (ngemuva kwalokhu okubizwa ngokuthi i-Cap1, Cap2, Cap3, ne-Cap4).I-fluorescence eyenziwe nge-laser esuka ku-Cap1-Cap4 yahlanganiswa nohlu lwamalensi amane futhi yaqoshwa kanyekanye nge-spectrometer enemibala eyisishiyagalolunye.I-intensity dynamics yemibala eyisishiyagalolunye (C1-C9) fluorescence ngesikhathi se-electrophoresis, okungukuthi, i-electrophoregram yemibala eyisishiyagalolunye ye-capillary ngayinye, iboniswa ku-Fig. 7a.I-electrophoregram yemibala eyisishiyagalolunye elinganayo itholakala ku-Cap1-Cap4.Njengoba kukhonjiswe imicibisholo ye-Cap1 ku-Figure 7a, iziqongo eziyisishiyagalombili ku-electrophoregram ngayinye enemibala eyisishiyagalolunye zibonisa ukukhishwa kwe-fluorescence okukodwa okuvela ku-Dye1-Dye8, ngokulandelana.
Ukulinganisa ngesikhathi esisodwa odayi abayisishiyagalombili kusetshenziswa i-spectrometer ye-electrophoresis enemibala eyisishiyagalolunye yemibala eyisishiyagalolunye.(a) I-electrophoregram yemibala eyisishiyagalolunye (C1-C9) ye-capillary ngayinye.Iziqongo eziyisishiyagalombili ezikhonjiswe imicibisholo i-Cap1 zibonisa ukukhishwa kwe-fluorescence ngakunye kodayi abayisishiyagalombili (Dye1-Dye8).Imibala yemicibisholo ihambisana nemibala (b) kanye no-(c).(b) I-Fluorescence spectra yodayi abayisishiyagalombili (Dye1-Dye8) nge-capillary ngayinye.c Ama-Electropherograms odayi abayisishiyagalombili (Dye1-Dye8) nge-capillary ngayinye.Iziqongo zezingcezu ze-DNA ezinelebula le-Dye7 ziboniswa ngemicibisholo, futhi ubude bazo besisekelo se-Cap4 buyaboniswa.
Ukusatshalaliswa kokuqina kwe-C1–C9 eziqongweni eziphakeme eziyisishiyagalombili kuboniswa ku-Fig.7b, ngokulandelana.Ngenxa yokuthi kokubili i-C1-C9 ne-Dye1-Dye8 zilandelana ngobude begagasi, ukusabalalisa okuyisishiyagalombili ku-Fig. 7b kubonisa i-spectra ye-fluorescence ye-Dye1-Dye8 ngokulandelana ukusuka kwesobunxele kuye kwesokudla.Kulolu cwaningo, i-Dye1, i-Dye2, i-Dye3, i-Dye4, i-Dye5, i-Dye6, i-Dye7, ne-Dye8 ivela ku-magenta, violet, blue, cyan, green, yellow, orange, and red, ngokulandelana.Qaphela ukuthi imibala yemicibisholo ekuFig. 7a ihambisana nemibala kadayi kuFig. 7b.Amandla e-C1-C9 e-fluorescence ku-spectrum ngayinye ku-Figure 7b aye ajwayelekile ukuze isamba sawo silingane nesisodwa.Izibukeli eziyisishiyagalombili ezilinganayo ze-fluorescence zitholwe ku-Cap1-Cap4.Umuntu angabona ngokucacile ukugqagqana kwe-spectral kwe-fluorescence phakathi kodayi ongu-1-dayi 8.
Njengoba kuboniswe kuMfanekiso 7c, ku-capillary ngayinye, i-electrophoregram yemibala eyisishiyagalolunye kuMfanekiso 7a yaguqulwa yaba i-electropherogram enodayi oyisishiyagalombili ngokuhlaziywa kwezingxenye eziningi okusekelwe ku-spectra ye-fluorescence eyisishiyagalombili ku-Figure 7b (bona Izinsiza Zokungezelela ukuze uthole imininingwane).Njengoba ukugqagqana kwe-spectral kwe-fluorescence ku-Figure 7a kungavezwa kuMfanekiso 7c, i-Dye1-Dye8 ingahlonzwa futhi ibalwe ngayodwa ngesikhathi ngasinye, ngisho noma amanani ahlukene e-Dye1-Dye8 fluoresce ngesikhathi esisodwa.Lokhu ngeke kwenziwe ngokutholwa kwemibala eyisikhombisa31 okungokwesiko, kodwa kungazuzwa ngokutholwa kwemibala eyisishiyagalolunye okuthuthukisiwe.Njengoba kuboniswa imicibisholo i-Cap1 ku-Fig. 7c, i-fluorescent emission singlets i-Dye3 (eluhlaza okwesibhakabhaka), i-Dye8 (ebomvu), i-Dye5 (eluhlaza okotshani), i-Dye4 (cyan), i-Dye2 (purple), i-Dye1 (magenta), ne-Dye6 (Ephuzi ) zibhekwa ngokulandelana okulindelekile.Ngokukhishwa kwe-fluorescent kadayi wesi-7 (owolintshi), ngaphezu kwesiqongo esisodwa esikhonjiswe umcibisholo owolintshi, ezinye iziqongo ezimbalwa ezimbalwa zabonwa.Lo mphumela ungenxa yokuthi amasampula aqukethe izilinganiso zosayizi, i-Dye7 ebhalwe izingcezu ze-DNA ezinobude obuhlukene besisekelo.Njengoba kuboniswe kuMfanekiso 7c, ku-Cap4 lobu bude besisekelo bungu-20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 kanye nobude besisekelo obungu-220.
Izici eziyinhloko ze-spectrometer yemibala eyisishiyagalolunye, ethuthukiswe kusetshenziswa i-matrix yezibuko ze-dichroic ezinezingqimba ezimbili, ziwusayizi omncane kanye nomklamo olula.Njengoba uxhaxha lwezibuko ze-decachromatic ngaphakathi kwe-adaptha ekhonjiswe ku-fig.I-3c ifakwe ngokuqondile ebhodini lenzwa yesithombe (bheka u-Fig. S1 no-S2), i-spectrometer yemibala eyisishiyagalolunye inobukhulu obufanayo ne-adaptha, okungukuthi 54 × 58 × 8.5 mm.(ubukhulu).Lo sayizi omncane kakhulu ungama-oda amabili kuya kwamathathu obukhulu amancane kunama-spectrometer avamile asebenzisa ama-gratings noma ama-prism.Ngaphezu kwalokho, njengoba i-spectrometer enemibala eyisishiyagalolunye imisiwe ngendlela yokuthi ukukhanya kushaye inzwa yesithombe ngokuyikho, isikhala singabelwa kalula i-spectrometer enemibala eyisishiyagalolunye ezinhlelweni ezifana nama-microscopes, ama-flow cytometer, noma izihlaziyi.I-Capillary grating electrophoresis analyzer ye-miniaturization enkulu nakakhulu yesistimu.Ngesikhathi esifanayo, ubukhulu bezibuko eziyishumi ze-dichroic nezihlungi ze-bandpass ezisetshenziswe ku-spectrometer yemibala eyisishiyagalolunye yi-10×1.9×0.5 mm kuphela noma 15×1.9×0.5 mm.Ngakho-ke, ngaphezu kwe-100 izibuko ezincane ze-dichroic nezihlungi ze-bandpass, ngokulandelanayo, zingasikwa esibukweni se-dichroic kanye nesihlungi se-bandpass esingu-60 mm2, ngokulandelana.Ngakho-ke, izibuko ze-decachromatic zingenziwa ngezindleko eziphansi.
Esinye isici se-spectrometer yemibala eyisishiyagalolunye yizici zayo ezinhle kakhulu ze-spectral.Ikakhulukazi, ivumela ukutholwa kwezithombe ze-spectral zezifinyezo, okungukuthi, ukutholwa kanyekanye kwezithombe ngolwazi lwe-spectral.Esithombeni ngasinye, i-spectrum eqhubekayo itholwe ngebanga le-wavelength ukusuka ku-520 kuya ku-700 nm kanye nesinqumo esingu-20 nm.Ngamanye amazwi, amandla okukhanya emibala ayisishiyagalolunye atholwa esithombeni ngasinye, okungukuthi amabhendi ayisishiyagalolunye angu-20 nm ahlukanisa ngokulinganayo ububanzi be-wavelength ukusuka ku-520 ukuya ku-700 nm.Ngokushintsha izici ze-spectral zesibuko se-dichroic nesihlungi se-bandpass, ububanzi be-wavelength bamabhande ayisishiyagalolunye nobubanzi bebhande ngalinye kungalungiswa.Ukutholwa kwemibala eyisishiyagalolunye ngeke kusetshenziselwe izilinganiso ze-fluorescence kuphela nge-spectral imaging (njengoba kuchazwe kulo mbiko), kodwa nakwezinye izinhlelo zokusebenza eziningi ezivamile ezisebenzisa i-spectral imaging.Nakuba imaging ye-hyperspectral ingathola amakhulu emibala, kutholakale ukuthi ngisho nokunciphisa okuphawulekayo kwenani lemibala ebonakalayo, izinto eziningi emkhakheni wokubuka zingabonakala ngokunemba okwanele kwezinhlelo zokusebenza eziningi38,39,40.Ngenxa yokuthi ukulungiswa kwendawo, ukulungiswa kwe-spectral, nokulungiswa kwesikhashana kune-tradeoff emfanekisweni we-spectral, ukwehlisa inani lemibala kungathuthukisa ukulungiswa kwendawo kanye nokulungiswa kwesikhashana.Ingase futhi isebenzise ama-spectrometer alula njengalena ethuthukiswe kulolu cwaningo futhi inciphise nakakhulu inani lokubala.
Kulolu cwaningo, odayi abayisishiyagalombili balinganiswa ngesikhathi esisodwa ngokuhlukaniswa kwe-spectral ye-spectra yabo egqagqene ye-fluorescence ngokusekelwe ekutholweni kwemibala eyisishiyagalolunye.Odayi abangafika kwabayisishiyagalolunye bangalinganiswa ngesikhathi esisodwa, bahlale ngesikhathi nendawo.Inzuzo ekhethekile ye-spectrometer enemibala eyisishiyagalolunye ukugeleza kwayo okukhanyayo okuphezulu kanye nendawo enkulu yokuvula (1 × 7 mm).I-decane mirror array inokudlulisa okuphezulu okungama-92% wokukhanya okuvela endaweni evulekile kububanzi obuyisishiyagalolunye be-wavelength.Ukusebenza kahle kokusebenzisa ukukhanya kwesigameko ebangeni le-wavelength kusuka ku-520 kuya ku-700 nm cishe ku-100%.Kulolu hlu olubanzi lwamaza wamaza, akukho grating ye-diffraction enganikeza ukusebenza kahle okuphezulu kangaka kokusetshenziswa.Ngisho noma ukusebenza kahle kokuphazamiseka kwe-diffraction grating kudlula ama-90% kubude begagasi obuthile, njengoba umehluko phakathi kwalolo bude beza beza nobude obuthile ukhula, ukusebenza kahle kokwehlukanisa kwelinye ubude beza kuncipha41.Ububanzi bembobo buncike ekuqondeni kwendiza ku-Fig. 2c bunganwetshwa kusuka ku-7 mm kuya kububanzi benzwa yesithombe, njengalapho kwenzeka inzwa yesithombe esetshenziswe kulolu cwaningo, ngokushintsha kancane uhlu lwe-decamer.
I-spectrometer yemibala eyisishiyagalolunye ingasetshenziswa hhayi kuphela kwi-capillary electrophoresis, njengoba kuboniswe kulolu cwaningo, kodwa futhi nangezinye izinjongo ezihlukahlukene.Isibonelo, njengoba kuboniswe emfanekisweni ongezansi, i-spectrometer enemibala eyisishiyagalolunye ingasetshenziswa kusibonakhulu se-fluorescence.Indiza yesampula iboniswa kunzwa yesithombe ye-spectrometer enemibala eyisishiyagalolunye ngomgomo we-10x.Ibanga lokubona phakathi kwelensi yenhloso kanye nenzwa yesithombe ngu-200 mm, kuyilapho ibanga lokubona phakathi kwendawo yesigameko ye-spectrometer enemibala eyisishiyagalolunye kanye nenzwa yesithombe ingu-12 mm kuphela.Ngakho-ke, isithombe sasikwa safinyelela cishe kusayizi wembobo (1 × 7 mm) endizeni yesigameko futhi sahlukaniswa saba yizithombe ezinemibala eyisishiyagalolunye.Okusho ukuthi, isithombe se-spectral sesithombe esifushane esinemibala eyisishiyagalolunye singathathwa endaweni engu-0.1×0.7 mm endizeni yesampula.Ngaphezu kwalokho, kungenzeka ukuthola isithombe se-spectral esinemibala eyisishiyagalolunye sendawo enkulu endizeni yesampula ngokuskena isampula ehlobene nenhloso endaweni evundlile ku-Fig. 2c.
Izingxenye ze-decachromatic mirror array, okuyi-M1-M9 ne-BP, zenziwe ngokwezifiso i-Asahi Spectra Co., Ltd. kusetshenziswa izindlela ezijwayelekile zemvula.Izinto zokwakha ze-Multilayer dielectric zafakwa ngazinye kumapuleti equartz ayishumi angu-60 × 60 mm ngobukhulu nogqinsi ongu-0.5 mm, okuhlangabezana nezidingo ezilandelayo: M1: IA = 45°, R ≥ 90% ku-520–590 nm, Tave ≥ 90% ku-610– 610 nm.700 nm, M2: IA = 45°, R ≥ 90% ku-520–530 nm, Tave ≥ 90% ku-550–600 nm, M3: IA = 45°, R ≥ 90% ku-540–550 nm, Tave 0 % ku-570–600 nm, M4: IA = 45°, R ≥ 90% ku-560–570 nm, Tave ≥ 90% ku-590–600 nm, M5: IA = 45°, R ≥ 98% ku-580-600 nm , R ≥ 98% ku-680–700 nm, M6: IA = 45°, Tave ≥ 90% ku-600–610 nm, R ≥ 90% ku-630–700 nm, M7: IA = 45°, R ≥ 90% 620–630 nm, Taw ≥ 90% ku-650–700 nm, M8: IA = 45°, R ≥ 90% ku-640–650 nm, Taw ≥ 90% ku-670–700 nm, M9: IA = 45° ≥ 90% ku-650-670 nm, Tave ≥ 90% ku-690-700 nm, BP: IA = 0 °, T ≤ 0.01% ku-505 nm, Tave ≥ 95% ku-530-690 nm T0≥ 530 nm ku-530 nm ku- -690 nm kanye ne-T ≤ 1% ku-725-750 nm, lapho i-IA, T, Tave, ne-R kuyi-engeli yesigameko, ukudluliswa, ukudluliswa kwesilinganiso, nokubonakaliswa kokukhanya okungahlukanisiwe.
Ukukhanya okumhlophe (C0) okunobubanzi begagasi obungu-400–750 nm obukhishwa umthombo wokukhanya we-LED (AS 3000, AS ONE CORPORATION) kwahlanganiswa futhi kwenzeka mpo ku-DP yezibuko ze-dichroic.I-spectrum yokukhanya okumhlophe kwama-LED akhonjiswa ku-Supplementary Figure S3.Beka ithangi le-acrylic (ubukhulu obuyi-150 × 150 × 30 mm) ngqo phambi kohlu lwesibuko se-decamera, maqondana ne-PSU.Intuthu ekhiqizwe lapho iqhwa elomile licwiliswa emanzini labe selithelwa ethangini le-acrylic ukuze kubonwe imifudlana ehlukanisayo yemibala eyisishiyagalolunye ye-C1-C9 ephuma ochungechungeni lwezibuko ze-decachromatic.
Okunye, ukukhanya okumhlophe okuhlanganisiwe (C0) kudluliselwa ngesihlungi ngaphambi kokungena ku-DP.Izihlungi ekuqaleni beziyizihlungi zokuminyana ezingathathi hlangothi ezine-optical density engu-0.6.Bese usebenzisa isihlungi esinenjini (FW212C, FW212C, Thorlabs).Ekugcineni, vula futhi isihlungi se-ND.I-bandwidths yezihlungi eziyisishiyagalolunye ze-bandpass zihambelana ne-C9, i-C8, i-C7, i-C6, i-C5, i-C4, i-C3, i-C2 ne-C1, ngokulandelanayo.Iseli le-quartz elinobukhulu bangaphakathi obungu-40 (ubude be-optical) x 42.5 (ubude) x 10 mm (ububanzi) labekwa phambi kohlu lwezibuko ze-decochromatic, ngokuphambene ne-BP.Intuthu ibe isifakwa ngeshubhu kuseli le-quartz ukuze kugcinwe ukugcwala kwentuthu kuseli le-quartz ukuze ubone ngeso lengqondo imifudlana ehlukanisayo yemibala eyisishiyagalolunye ye-C1-C9 ephuma kuhlu lwesibuko se-decachromatic.
Ividiyo yokusakaza kokukhanya okuhlukanisayo okunemibala eyisishiyagalolunye ephuma kuxhaxha lwezibuko ze-decanic yathwetshulwa ngemodi yokudlula isikhathi ku-iPhone XS.Thwebula izithombe zesigcawu ngo-1 fps futhi uhlanganise izithombe ukuze udale ividiyo ku-30 ​​fps (ngevidiyo ozikhethela yona 1) noma 24 fps (ngamavidiyo ozikhethela 2 no-3).
Beka ipuleti lensimbi engagqwali eliwugqinsi elingu-50 µm (elinezimbobo ezine zobubanzi obungu-50 µm ngezikhathi ezingu-1 mm) epuleti le-diffusion.Ukukhanya okunobude begagasi obungu-400-750 nm kufakwa imisebe epuleti le-diffuser, okutholwe ngokudlula ukukhanya kwesibani se-halogen ngesihlungi esifushane sokudlulisa esinobude obungamaza obungama-700 nm.I-spectrum yokukhanya iboniswa ku-Supplementary Figure S4.Kungenjalo, ukukhanya kuphinde kudlule kwesinye sezihlungi ze-10 nm bandpass ezigxile ku-530, 550, 570, 590, 610, 630, 650, 670 kanye ne-690 nm bese kushaya ipuleti le-diffuser.Ngenxa yalokho, amaphoyinti emisebe amane anobubanzi obungu-φ50 μm namaza amaza ahlukene akhiwe epuleti lensimbi engagqwali elibhekene nepuleti le-diffuser.
Uhlu lwama-capillary amane olunamalensi amane lufakwe ku-spectrometer enemibala eyisishiyagalolunye njengoba kuboniswe kuMfanekiso 1 no-2. C1 kanye ne-C2.Ama-capillary amane namalensi amane ayefana nezifundo zangaphambili31,34.I-laser beam ene-wavelength engu-505 nm namandla angu-15 mW ifakwe imisebe kanye kanye nangokulinganayo isuka eceleni iye ezindaweni ezikhipha ama-capillaries amane.I-fluorescence ekhishwa iphoyinti ngalinye lokuphuma ihlanganiswa ilensi ehambisanayo futhi ihlukaniselwe imifudlana yemibala eyisishiyagalolunye ngoxhaxha lwezibuko ze-decachromatic.Umphumela wokusakaza ongu-36 wabe usujovwa ngokuqondile kunzwa yesithombe se-CMOS (C11440–52U, Hamamatsu Photonics K·K.), futhi izithombe zabo zarekhodwa kanyekanye.
I-ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Applied Biosystems), 4 µl GeneScan™ 600 LIZ™ idayi yaxutshwa ku-capillary ngayinye ngokuxuba i-1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), usayizi wemiksi engu-1 µl ojwayelekile.v2.0 (Thermo Fisher Scientific) kanye no-14 µl wamanzi.I-PowerPlex® 6C Matrix Standard iqukethe izingcezu eziyisithupha ze-DNA ezilebulwe ngodayi abayisithupha: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, kanye ne-WEN, ngokulandelana kobude begagasi obukhulu.Ubude besisekelo balezi zingcezu ze-DNA abudalulwa, kodwa ukulandelana kobude besisekelo kwezingcezu ze-DNA ezilebulwe ngokuthi WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C kanye ne-TOM-6C kuyaziwa.Ingxube ku-ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit iqukethe ucezu lwe-DNA olulebulwe ngodayi we-dR6G.Ubude bezisekelo zezingcezwana ze-DNA nabo abudalulwa.I-GeneScan™ 600 LIZ™ Usayizi Wedayi Okujwayelekile i-v2.0 ihlanganisa izingcezu ze-DNA ezinelebuli ye-LIZ.Ubude obuyisisekelo balezi zingcezu ze-DNA bungama-20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 40, 30, 30 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 kanye nesisekelo sama-600.Amasampula afakwe ku-94 ° C imizuzu engu-3, ​​bese epholiswa eqhweni imizuzu emi-5.Amasampula afakwe ku-capillary ngayinye ngo-26 V/cm ngamasekhondi angu-9 futhi ahlukaniswa ku-capillary ngayinye egcwele isixazululo se-polymer ye-POP-7™ (Thermo Fisher Scientific) enobude obusebenzayo obungu-36 cm kanye nogesi ongu-181 V/cm i-engeli engu-60°.KUSUKA.
Yonke idatha etholiwe noma ehlaziywe phakathi nalolu cwaningo ifakiwe kulesi sihloko esishicilelwe kanye nolwazi lwaso olwengeziwe.Eminye imininingwane ehlobene nalolu cwaningo iyatholakala kubabhali abafanele uma kunesicelo esifanele.
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