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Qalab is haysta (54 × 58 × 8.5 mm) iyo dalool ballaadhan (1 × 7 mm) spectrometer sagaal midab leh ayaa la sameeyay, “laba u kala qaybiyey” oo ay soo diyaariyeen toban muraayadood oo dichroic ah, kuwaas oo loo adeegsaday sawir-muuqaal degdeg ah.Iftiinka dhacdada oo leh qayb iskutallaab ah oo ka yar cabbirka daloolka waxa loo qaybiyaa xariijin joogto ah oo ballac ah 20 nm iyo sagal qulqulo midab leh oo leh hirarka dhexda ee 530, 550, 570, 590, 610, 630, 650, 670 iyo 690 nm.Sawirada sagaal ilbiriqsi oo midab leh ayaa isku mar si hufan loogu cabbiraa dareenka sawirka.Si ka duwan sida muraayadaha muraayadaha caadiga ah, muraayadda muraayadda dichroic ee la horumariyay waxay leedahay qaabeynta laba qaybood oo gaar ah, taas oo aan kordhinayn tirada midabada isku mar la qiyaasi karo, laakiin sidoo kale waxay hagaajinaysaa xallinta sawirka midab kasta.spectrometer-ka la sameeyay ee sagaal midab leh waxaa loo isticmaalaa korantada afar-capillary.Falanqaynta tiro isku mar ah oo siddeed midabyo ah oo isku mar u guuraya xidid kasta iyada oo la adeegsanayo fluorescence sagaal midab leh oo leysarka ah.Maadaama spectrometer-ka sagaal-midabku aanu ahayn oo keliya mid aad u yar oo aan qaali ahayn, laakiin sidoo kale uu leeyahay qulqul iftiin sare leh iyo xallin ku filan inta badan codsiyada sawir-qaadista, waxaa si ballaaran loo isticmaali karaa meelo kala duwan.
Muuqaal-muuqaal sare iyo muuqaalo kala duwan ayaa noqday qayb muhiim ah oo ka mid ah xiddigiska2, dareenka fog ee indha-indheynta Dhulka3,4, xakamaynta tayada cuntada iyo biyaha5,6, ilaalinta farshaxanka iyo archeology7, forensics8, qaliin9, falanqaynta biomedical iyo ogaanshaha10,11 iwm. Field 1 Farsamo lama huraan ah ,12,13.Hababka lagu cabbiro xajmiga iftiinka ee ka soo baxa goob kasta oo ka mid ah qiiqa ka baxa goobta aragtida waxa loo qaybiyaa (1) sawir-qaadis ("xaaqin") , (3) dhererka sawirka mowjadaha19,20,21 iyo (4) images22,23,24,25.Marka la eego hababkan oo dhan, xallinta meelaha, xallinta muuqaalka iyo xallinta ku meel gaarka ah waxay leeyihiin xiriir ganacsi-off9,10,12,26.Intaa waxaa dheer, soosaarka iftiinka ayaa saameyn weyn ku leh dareenka, tusaale ahaan saamiga calaamad-ilaa-sanqa ee sawirka muuqaalka26.Qulqulka iftiinka, taas oo ah, waxtarka isticmaalka iftiinka, ayaa si toos ah u dhigma saamiga dhabta ah ee qiyaasta qiyaasta iftiinka ee dhibic kasta oo iftiin ah halkii cutubba ilaa wadarta guud ee iftiinka qiyaasta hirarka dhererka.Qaybta (4) waa hab ku habboon marka xoogga ama xajmiga iftiinka ay ka baxayaan meel kasta oo bixisa ay is beddesho waqti ama marka meel kasta oo wax bixisa ay is beddesho waqtiga sababtoo ah spectrum iftiinka ee ay soo baxaan dhammaan dhibcuhu waa isku mar.24.
Inta badan hababka kor ku xusan waxaa lagu daraa spectrometers waaweyn, adag iyo/ama qaali ah iyadoo la isticmaalayo 18 gratings ama 14, 16, 22, 23 prisms ee fasalada (1), (2) iyo (4) ama 20, 21 saxanadaha filter, dareere filtarrada. .Shaandheeyayaasha kristal tunable (LCTF)25 ama acousto-optic tunable filtarrada (AOTF)19 ee qaybta (3).Taas bedelkeeda, qaybta (4) muraayadaha muraayadaha badan waa kuwo yaryar oo aan qaali ahayn sababtoo ah qaabayntooda fudud27,28,29,30.Intaa waxaa dheer, waxay leeyihiin iftiin sare oo iftiin leh sababtoo ah iftiinka ay wadaagaan muraayad kasta oo dichroic ah (taas oo ah, gudbinta iyo iftiinka iftiinka dhacdada ee muraayad kasta oo dichroic ah) ayaa si buuxda oo joogto ah loo isticmaalaa.Si kastaba ha ahaatee, tirada xadhkaha hirarka dhererka (ie midabada) ee ay tahay in isku mar la cabbiro waxay ku kooban tahay ilaa afar.
Sawirka Spectral ee ku salaysan ogaanshaha fluorescence waxaa caadi ahaan loo isticmaalaa falanqaynta multiplex ee ogaanshaha biomedical iyo ogaanshaha 10, 13.Isku dhufashada, maadaama falanqaynta badan (tusaale, DNA gaar ah ama borotiinno) lagu calaamadeeyay midabyo kala duwan oo fluorescent ah, falanqeeye kasta oo jooga meel kasta oo ka sii daayo goobta aragtida ayaa lagu qiyaasaa iyadoo la adeegsanayo falanqaynta qaybo badan.32 wuxuu jebiyaa spectrum fluorescence ee la ogaaday ee ay sii daysay bar kasta.Inta lagu jiro hawshan, midabyo kala duwan, oo mid kastaa soo saarayo iftiin kale, ayaa midabo kara, taas oo ah, wada noolaanshaha goobta iyo waqtiga.Waqtigan xaadirka ah, tirada ugu badan ee midabaynta ee uu ku farxi karo hal laydh oo kaliya ayaa ah siddeed33.Xadkan sare laguma go'aamiyo xallinta spectral (ie, tirada midabada), laakiin waxaa lagu qiyaasaa ballaca spectrum fluorescence (≥50 nm) iyo cadadka dheeha Stokes shift (≤200 nm) ee FRET (adoo isticmaalaya FRET)10 .Si kastaba ha ahaatee, tirada midabada waa in ay ka badan tahay ama la mid tahay tirada dheeha si loo baabi'iyo isku dhafka midabada isku dhafan31,32.Sidaa darteed, waxaa lagama maarmaan ah in la kordhiyo tirada midabada isku mar la qiyaasay ilaa siddeed ama ka badan.
Dhowaan, ultra-compact heptachroic spectrometer (iyadoo la isticmaalayo isku dubarid muraayado heptychroic ah iyo dareeraha sawirka si loo cabbiro afar fluorescent fluxes) ayaa la sameeyay.spectrometer-ku waa laba ilaa saddex amar oo cabbir ka yar spectrometer-yada caadiga ah iyadoo la isticmaalayo gratings ama prisms34,35.Si kastaba ha ahaatee, way adag tahay in la dhigo in ka badan toddoba muraayadood oo dichroic ah spectrometer oo isku mar la cabbiro in ka badan toddoba midab36,37.Kordhinta tirada muraayadaha dichroic, farqiga ugu sarreeya ee dhererka waddooyinka indhaha ee qulqulaha iftiinka dichroic ayaa kordha, wayna adkaanaysaa in lagu muujiyo dhammaan iftiinka iftiinka hal diyaarad dareen ah.Dhererka dariiqa indhaha ee ugu dheer ee qulqulka iftiinka ayaa sidoo kale kor u kaca, markaa ballaca daloolka spectrometerka (tusaale ballaca ugu badan ee iftiinka lagu falanqeeyay spectrometer) ayaa hoos u dhacaya.
Iyada oo laga jawaabayo dhibaatooyinka kor ku xusan, spectrometer ultra-compact ah oo sagaal midab leh oo leh laba lakab oo "dichroic" muraayado decachromatic ah iyo sawir sawir ah oo loogu talagalay sawirka muuqaalka degdega ah [qaybta (4)] ayaa la sameeyay.Marka la barbar dhigo spectrometer-yadii hore, spectrometer-ka la horumariyay wuxuu leeyahay farqi yar oo dhererka jidka ugu sarreeya iyo dhererka dariiqa indhaha ee ugu yar.Waxaa lagu dabaqay afar-xajiseed electrophoresis si loo ogaado laysarka sagal-midab leh fluorescence iyo in la qiyaaso guuritaanka isku mar ah ee siddeed midab ee xidid kasta.Maadaama spectrometer-ka la horumariyay aanu ahayn mid aad u yar oo aan qaali ahayn, laakiin sidoo kale uu leeyahay qulqul iftiin sare leh iyo xallin ku filan inta badan codsiyada sawirada muuqaalka ah, waxaa si ballaaran loo isticmaali karaa meelo kala duwan.
spectrometer-ka dhaqanka ee sagaal-midabka ah waxa lagu muujiyay berdaha.1a.Naqshaddeedu waxay raacdaa tii hore ee ultra-yar-yar oo toddoba midab leh 31. Waxay ka kooban tahay sagaal muraayado dichroic ah oo si siman u habaysan xagal 45 ° dhinaca midig ah, iyo sawirka sawirka (S) wuxuu ku yaalaa korka sagaal muraayadaha dichroic.Iftiinka ka soo galaya xagga hoose (C0) waxa loo qaybiyaa sagal muraayado dichroic ah oo kala duwan oo u qaybiya sagaal iftiin oo kor u socda (C1, C2, C3, C4, C5, C6, C7, C8 iyo C9).Dhammaan sagaalka durdurrada midabka waxaa si toos ah loogu quudiyaa dareeraha sawirka waxaana la ogaanayaa isku mar.Daraasaddan, C1, C2, C3, C4, C5, C6, C7, C8, iyo C9 waxay u kala horreeyaan mowjadaha dhererka waxayna u taagan yihiin magenta, violet, buluug, cyan, cagaar, jaalle, oranji, casaan-orange, iyo cas, siday u kala horreeyaan.In kasta oo calaamadahan midabada ah lagu isticmaalo dukumeentigan, sida ku cad sawirka 3, sababtoo ah waxay ka duwan yihiin midabada dhabta ah ee ay arkaan isha bini'aadamka.
Jaantusyada jaantusyada spectrometer-ka cusub ee caadiga ah iyo kuwa cusub.(a) spectrometer-ka caadiga ah ee sagaal midab leh oo leh isku dubaridan sagaal muraayadood oo dichroic ah.(b) spectrometer cusub oo sagaal midab leh oo leh muraayad laba lakab ah.Dhacdada iftiinka iftiinka C0 waxa loo qaybiyaa sagaal ilbiriqsi oo midab leh C1-C9 waxaana lagu ogaaday dareenka sawirka S.
spectrometer-ka cusub ee sagaal midab leh ee la sameeyay waxa uu leeyahay shaadhka muraayada laba-lakab ee dichroic iyo dareeraha sawirka, sida ku cad sawirka 1b.Heerka hoose, shan muraayadood oo dichroic ah ayaa u janjeera 45° dhanka midig, oo ku toosan dhanka midig oo ka bilaabmaya dhexda isku xidhka qalabyada.Heerka sare, shan muraayadood oo dheeraad ah oo dichroic ah ayaa u janjeera 45 ° bidix waxayna ku yaalliin bartamaha ilaa bidix.Muraayada dichroic-ka bidix ee lakabka hoose iyo muraayadda dichroic-ka midig ee lakabka sare ayaa midba midka kale isku dhejiyaa.Iftiinka iftiinka dhacdada (C0) wuxuu hoos u kala qaybsan yahay afar dareere koromotik ah (C1-C4) oo ay wataan shan muraayadood oo dichroic ah oo midigta ah iyo shanta dareeraha chromatic fluxes (C5-C4) ee shan muraayadaha dichroic ee bidixda C9).Sida spectrometer-ka caadiga ah ee sagaal-midabka ah, dhammaan sagaalka durdurrada midabka si toos ah ayaa loogu duraa dareenka sawirka (S) waxaana la ogaanayaa isku mar.Isbarbardhigga jaantusyada 1a iyo 1b, midku wuxuu arki karaa in marka laga hadlayo spectrometer-ka cusub ee sagaal-midabka ah, labadaba farqiga ugu badan iyo dhererka dariiqa indhaha ee ugu dheer ee sagaasha midabada midabku waa kala badh.
Dhismaha faahfaahsan ee muraayadda laba-lakab ee aadka u yar-yar ee 29 mm (ballaaran) × 31 mm (dherer) × 6 mm (dherer) ayaa lagu muujiyey sawirka 2. Qalabka muraayadda jajab tobanle wuxuu ka kooban yahay shan muraayadood oo dichroic ah oo midigta ah. (M1-M5) iyo shan muraayadood oo dichroic ah oo bidixda ah (M6-M9 iyo M5 kale), muraayad kasta oo dichroic ah ayaa lagu dhejiyaa garabka sare ee aluminium.Dhammaan muraayadaha dichroic waa la rog-rogay si loo magdhabo barokaca barbar socda sababtoo ah qulqulka qulqulka muraayadaha.Hooska M1, shaandhada band-pass (BP) ayaa go'an.M1 iyo cabbirada BP waa 10mm (dhinaca dheer) x 1.9mm (dhinaca gaaban) x 0.5mm ( dhumucdiisuna tahay ).Cabbirrada muraayadaha dichroic ee soo hadhay waa 15 mm × 1.9 mm × 0.5 mm.Dhererka shaxanka ee u dhexeeya M1 iyo M2 waa 1.7 mm, halka cabbirka cabbirka ee muraayadaha kale ee dichroic uu yahay 1.6 mm.On berde.2c wuxuu isku daraa qulqulka iftiinka dhacdada C0 iyo sagaal ilbiriqsiyo midab leh C1-C9, oo ay kala soocaan matrix-ka-qolka muraayadaha.
Dhismaha muraayad laba-lakab ah oo dichroic ah.(a) Aragtida aragtida iyo (b) aragti iskutallaab ah oo ah muraayad laba lakab ah oo isku dhafan (cabir 29 mm x 31 mm x 6 mm).Waxay ka kooban tahay shan muraayadood oo dichroic ah (M1-M5) oo ku yaal lakabka hoose, shan muraayadood oo dichroic ah (M6-M9 iyo M5 kale) oo ku yaal lakabka sare, iyo filter filter (BP) oo ku yaal meel ka hooseeya M1.(c) Aragtida isdhaafka ah ee jihada toosan, oo leh C0 iyo C1-C9 oo is dulsaaran.
Balaadhka daloolka ee jihada toosan, ee uu tilmaamayo ballaca C0 ee sawirka 2, c, waa 1 mm, iyo jihada toosan ee diyaaradda sawirka 2, c, oo lagu bixiyo naqshadaynta garabka aluminium. - 7 mm.Taasi waa, spectrometer-ka cusub ee sagaal midab leh wuxuu leeyahay cabbirka daloolka weyn ee 1 mm × 7 mm.Waddada indhaha ee C4 waa tan ugu dheer C1-C9, iyo dariiqa indhaha ee C4 ee gudaha muraayadda muraayadda dichroic, sababtoo ah cabbirka aadka u yar ee kor ku xusan (29 mm × 31 mm × 6 mm), waa 12 mm.Isla mar ahaantaana, dhererka jidka indhaha ee C5 waa kan ugu gaaban C1-C9, iyo dhererka dariiqa indhaha ee C5 waa 5.7mm.Sidaa darteed, farqiga ugu badan ee dhererka wadada indhaha waa 6.3 mm.Dhererka dariiqyada indhaha ee kor ku xusan ayaa lagu saxay dhererka jidka indhaha ee gudbinta indhaha ee M1-M9 iyo BP (laga bilaabo quartz).
Qalabka muuqaalka ah ee М1-М9 iyo VR waxaa loo xisaabiyaa si qulqulka С1, С2, С3, С4, С5, С6, С7, С8 iyo С9 ay ku dhex jiraan mawjadaha dhererka 520-540, 540-560, 560-580, -600, 600-620, 620-640, 640-660, 660-680, iyo 680-700 nm, siday u kala horreeyaan.
Sawirka shaxanka la soo saaray ee muraayadaha decachromatic ayaa lagu muujiyay sawirka 3a.M1-M9 iyo BP waxay ku dheggan yihiin jiirada 45-degree iyo diyaaradda toosan ee taageerada aluminiumka, siday u kala horreeyaan, halka M1 iyo BP ay ku qarsoon yihiin gadaasha sawirka.
Soo saarida isku dubaridan muraayadaha decan iyo muujinteeda.(a) Muraayadyo kala duwan oo la sameeyay.(b) 1 mm × 7 mm sawir kala qaybsan oo sagaal midab leh oo lagu saadaaliyay xaashi la hor dhigay isku dubaridan muraayadaha dekachromatic iyo dib u iftiiminaya iftiin cad.(c) Muraayadyo kala duwan oo qurxin leh oo iftiin cad gadaal ka yimid.(d) Sagaal-midab kala qaybsanaan oo ka soo baxa muraayadda muraayadda, oo lagu arkay iyada oo la gelinayo daasad akrilik ka buuxo oo qiiq ka buuxo oo la hor dhigay safafka muraayadda decane ee c oo madoobaynaya qolka.
Qiyaasta gudbinta la qiyaasay ee M1-M9 C0 ee xagasha dhacdooyinka 45 ° iyo qiyaasta gudbinta qiyaasta ee BP C0 ee xagasha dhacdooyinka 0 ° ayaa lagu muujiyay Fig.4a.Muuqaalka gudbinta ee C1-C9 marka loo eego C0 waxa lagu muujiyay berdihii.4b.Muuqaaladan waxaa laga soo xisaabiyay spectra ee ku jira berdihii.4a si waafaqsan dariiqa indhaha ee C1-C9 ee sawirka 4a.1b iyo 2c.Tusaale ahaan, 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)], halkay TS (X) iyo [1 - TS (X)] waa gudbinta iyo milicsiga spectra X, siday u kala horreeyaan.Sida ku cad Jaantuska 4b, xadhkaha xadhkaha (bandwidth ≥50%) ee C1, C2, C3, C4, C5, C6, C7, C8 iyo C9 waa 521-540, 541-562, 563-580, 581-602, 603 -623, 624-641, 642-657, 659-680 iyo 682-699 nm.Natiijooyinkani waxay la socdaan heerarka la sameeyay.Intaa waxaa dheer, waxtarka isticmaalka iftiinka C0 waa mid sarreeya, taas oo ah, celceliska ugu sarreeya ee C1-C9 gudbinta iftiinka waa 92%.
Muuqaalka gudbinta muraayadda dichroic iyo qulqulka sagaal midab ee kala qaybsan.(a) M1-M9 oo la qiyaasay 45° dhacdo ah iyo BP oo ah 0° dhacdo.(b) Muuqaalka gudbinta ee C1-C9 marka loo eego C0 laga soo xisaabiyay (a).
On berde.3c, isku dubaridka muraayadaha dichroic waxay ku yaalaan si toosan, sidaas darteed dhinaceeda midig ee Jaantuska 3a waa dhinaca sare iyo iftiinka cad ee LED-ka isku-dhafka ah (C0) ayaa dib u iftiimaya.Isku dhafka muraayadaha decachromatic ee lagu muujiyay sawirka 3a waxaa lagu dhejiyay 54 mm (dherer) × 58 mm ( qoto dheer) × 8.5 mm ( dhumucdiisuna)On berde.3d, marka lagu daro gobolka ka muuqda fig.3c, haan akrilik ka buuxo oo qiiq ka buuxo ayaa la hor dhigay muraayado badan oo qurxin ah, iyadoo nalalka qolka la demiyay.Natiijo ahaan, sagaal durdurro dichroic ah ayaa ka dhex muuqda haanta, kuwaas oo ka soo baxaya muraayado kala duwan.Qulqul kastaa wuxuu leeyahay qayb iskutallaabta leydi leh oo cabbirkeedu dhan yahay 1 × 7 mm, taas oo u dhiganta cabbirka daloolka spectrometer-ka cusub ee sagaal midab leh.Jaantuska 3b, xaashida xaashida ah ayaa la hor dhigayaa isku dubbaridka muraayadaha dichroic ee Jaantuska 3c, iyo sawirka 1 x 7 mm ee sagaal durdurrada dichroic ee lagu saadaaliyay warqadda ayaa laga arkayaa jihada dhaqdhaqaaqa warqadda.durdurrada.Sagaalka midab kala soocida durdurrada berdaha.3b iyo d waa C4, C3, C2, C1, C5, C6, C7, C8 iyo C9 kor ilaa hoos, kuwaas oo sidoo kale laga arki karo jaantusyada 1 iyo 2. 1b iyo 2c.Waxaa lagu arkay midabyo u dhigma hirarkooda.Sababtoo ah xoogga iftiinka cad ee hooseeya ee LED (eeg sawirka Dheeraadka ah. S3) iyo dareenka kamarada midabka ee loo isticmaalo in lagu qabto C9 (682-699 nm) ee sawirka. Socodyada kale ee kala qaybsanaanta ayaa daciif ah.Si la mid ah, C9 si liidata ayaa loogu arki jiray isha qaawan.Dhanka kale, C2 (qulqulka labaad ee sare) wuxuu u muuqdaa cagaar Jaantuska 3, laakiin wuxuu u muuqdaa jaale badan isha qaawan.
Kala-guurka Jaantuska 3c ilaa d waxa lagu muujiyay Fiidiyowga Dheeraadka ah 1. Isla markiiba ka dib markii iftiinka cad ee LED-ku uu dhex maro shaxanka muraayadda decachromatic, waxay isku mar u kala baxdaa sagaal durdurro midab leh.Dhammaadkii, qiiqa tamarku si tartiib tartiib ah ayuu kor ilaa hoose uga soo baxay, sidaas darteed sagaal budo ah oo midab leh ayaa sidoo kale la waayay kor ilaa hoos.Taas bedelkeeda, Fiidiyowga Dheeriga ah 2, markii hirarka dhererka qulqulka iftiinka ee isku dhafka muraayadaha decachromatic laga beddelay mid dheer ilaa gaaban sida 690, 671, 650, 632, 610, 589, 568, 550 iyo 532 nm ., Kaliya kuwa u dhigma ee kala qaybsan ee sagaalka durdurrada kala qaybsan sida C9, C8, C7, C6, C5, C4, C3, C2, iyo C1 ayaa la soo bandhigay.Kaydka akril waxaa lagu beddelaa barkad quartz ah, iyo jeexjeexyada qulqulka qulqulaya ayaa si cad looga arki karaa jihada kor u kacda.Intaa waxaa dheer, muuqaal-hoosaadka 3 waa la tafatiray si ay qaybta isbeddelka mawjada ee qayb-hoosaadka 2 dib loo daawado.Tani waa tibaaxaha ugu hadal-haynta badan ee sifooyinka muraayadaha muraayadaha ee decochromatic.
Natiijooyinka kor ku xusan waxay muujinayaan in muraayadda muraayadda decachromatic ee la soo saaray ama muraayadda cusub ee sagaal midab leh ay u shaqeyso sidii loogu talagalay.spectrometer-ka cusub ee sagaal midab leh ayaa lagu sameeyay iyadoo lagu rakibayo isku dubaridan muraayado decachromatic ah oo leh adapters si toos ah looxa dareeme sawirka.
Qulqulka iftiinka leh ee dhererkiisu yahay 400 ilaa 750 nm, oo ay sii daayaan afar dhibcood oo shucaac ah φ50 μm, oo ku yaala 1 mm kala-duwanaanshaha jihada jaantuska ee diyaaradda 2c, siday u kala horreeyaan Cilmi-baarista 31, 34. Qalabka afar-lens wuxuu ka kooban yahay afar lenses φ1 mm oo leh dhumucdiisuna tahay 1.4 mm iyo garoon 1 mm.Afarta durdurrada is-daba-joogga ah (afar C0) ayaa ku dhacay DP-ga spectrometer cusub oo sagaal midab leh, oo u dhexeeya 1 mm.Muraayadaha kala duwan ayaa u qaybiya durdur kasta (C0) oo u qaybiya sagal midab leh (C1-C9).Natiijadu waxay tahay 36 durdur (afar qaybood oo C1-C9 ah) ayaa markaa si toos ah loogu duraa dareeraha sawirka CMOS (S) ee si toos ah ugu xidhan muraayado badan oo dichroic ah.Natiijo ahaan, sida ku cad sawirka 5a, sababtoo ah farqiga yar ee ugu sarreeya ee indhaha iyo dariiqa gaaban ee ugu sarreeya, sawirada dhammaan 36 durdurrada ayaa la ogaaday si isku mid ah oo cad oo leh cabbir isku mid ah.Marka loo eego muuqalada hoose (eeg Jaantuska Dheeraadka ah S4), xoogga sawirka afarta kooxood ee C1, C2 iyo C3 waa mid hooseeya.Lix iyo soddon sawir ayaa cabbirkoodu ahaa 0.57 ± 0.05 mm (macnaha ± SD).Haddaba, weynaynta sawirku waxa uu celcelis ahaan ahaa 11.4.Kala fogaanshaha tooska ah ee u dhexeeya sawiradu waa celcelis ahaan 1 mm (isku-fognaan la mid ah sida array lens) iyo kala dheeraynta toosan celcelis ahaan 1.6 mm (fog la mid ah sida muraayadda muraayadda dichroic).Sababtoo ah cabbirka sawirku aad ayuu uga yar yahay masaafada u dhexeysa sawirrada, sawir kasta waxaa loo cabbiri karaa si madax bannaan (oo leh isgoys hoose).Dhanka kale, sawirada siddeed iyo labaatanka durdur ee lagu duubay spectrometer-ka caadiga ah ee toddobada midab ee loo adeegsaday daraasaddeennii hore ayaa lagu muujiyay sawirka 5 B. Iskudubbada toddobada muraayadood ee dichroic ayaa la abuuray iyada oo laga saaray labada muraayadood ee saxda ah ee saxda ah ee sagaal dichroic. muraayadaha sawirka 1a.Dhammaan sawiradu maaha kuwo fiiqan, cabbirka sawirku wuxuu ka korayaa C1 ilaa C7.Labaatan iyo siddeed sawir ayaa cabbirkoodu yahay 0.70 ± 0.19 mm.Sidaa darteed, way adag tahay in la ilaaliyo xallinta sawirka sare ee sawirada oo dhan.Isku-dhafka kala duwanaanshaha (CV) ee cabbirka sawirka 28 ee Jaantuska 5b wuxuu ahaa 28%, halka CV-ga cabbirka sawirka 36 ee Jaantuska 5a uu hoos ugu dhacay 9%.Natiijooyinka kor ku xusan waxay muujinayaan in spectrometer-ka cusub ee sagaal-midabka ah uusan kordhinaynin tirada midabada isku mar la qiyaaso min toddobo ilaa sagaal, laakiin sidoo kale wuxuu leeyahay xallin sare oo muuqaal ah midab kasta.
Isbarbardhigga tayada sawirka kala qaybsan ee ay sameeyeen spectrometer-yada caadiga ah iyo kuwa cusub.(a) Afar kooxood oo ah sawirro kala soocan oo sagaal midab leh (C1-C9) oo uu sameeyay spectrometer-ka cusub ee sagaal-midabka ah.(b) Afar qaybood oo ah sawirro toddoba midab leh (C1-C7) oo lagu sameeyay spectrometer toddoba midab leh.Fluxes (C0) oo leh mowjadaha dhererka laga bilaabo 400 ilaa 750 nm laga bilaabo afar dhibcood oo qiiqa ah ayaa la isku daraa oo dhacdo spectrometer kasta, siday u kala horreeyaan.
Sifooyinka spectrometer-ka sagaal-midabka ah ayaa lagu qiimeeyay si tijaabo ah natiijooyinka qiimeynta ayaa lagu muujiyay sawirka 6. Ogsoonow in Jaantuska 6a uu muujinayo natiijooyin isku mid ah jaantuska 5a, ie marka la eego mawjadaha 4 C0 400-750 nm, dhammaan sawirada 36 waa la ogaaday. (4 kooxood C1-C9).Taa lidkeeda, sida ku cad sawirka 6b-j, marka C0 kasta uu leeyahay mawjadda dhererka gaarka ah ee 530, 550, 570, 590, 610, 630, 650, 670, ama 690 nm, waxaa jira ku dhawaad afar sawir oo u dhigma (afar sawir). kooxuhu waxay ogaadeen C1, C2, C3, C4, C5, C6, C7, C8 ama C9).Si kastaba ha ahaatee, qaar ka mid ah sawirada ku dheggan afarta sawir ee u dhigma ayaa si aad u liidata loo ogaaday sababtoo ah C1-C9 gudbinta spectra ee lagu muujiyay shaxanka.Natiijooyinkani waxay la socdaan muuqaalka gudbinta C1-C9 ee lagu muujiyey berdihii.4b iyo fiidiyowyo dheeri ah 2 iyo 3. Si kale haddii loo dhigo, spectrometer-ka midabka ee sagaalku wuxuu u shaqeeyaa sidii la filayay isagoo ku saleysan natiijooyinka lagu muujiyey berdaha.4b.Sidaa darteed, waxaa lagu soo gabagabeeyay in qaybinta xoojinta sawirka C1-C9 ay tahay xajmiga C0 kasta.
Sifooyinka spectrometer ee sagaal midab leh.Qiyaasta cusub ee sagaal-midabka leh waxay soo saartaa afar qaybood oo ah sawirro sagaal midab leh (C1-C9) marka iftiinka shilka (afar C0) uu leeyahay mawjada dhererka (a) 400-750 nm (sida ku cad sawirka 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, siday u kala horreeyaan.
spectrometer-ka la sameeyay ee sagaal midab leh ayaa loo adeegsaday korantada afar-xabdeed (faahfaahinta, eeg Qalabka Dheeraadka ah)31,34,35.Matrixka afar-xabeedka ah wuxuu ka kooban yahay afar xabbo (dhexroorka dibadda ah 360 μm iyo dhexroorka gudaha 50 μm) oo ku yaalla 1 mm inta u dhaxaysa goobta laysarka.Tusaalooyinka ka kooban jajabyada DNA-da ee lagu calaamadeeyay 8 midabyo, kuwaas oo kala ah FL-6C (dheey 1), JOE-6C (dheee 2), dR6G (dheey 3), TMR-6C (dheey 4), CXR-6C (dheey 5), TOM- 6C (dheey 6), LIZ (dheeb 7), iyo WEN (dheey 8) oo u koraya siday u kala horreeyaan ee hirarka dhaadheer ee fluorescent, oo u kala qaybsan mid kasta oo ka mid ah afarta qaybood (markaan ka dib waxaa loo yaqaan Cap1, Cap2, Cap3, iyo Cap4).Fluorescence ka dhashay Laser-ka Cap1-Cap4 waxaa la isku daray dhowr lenses oo isku mar lagu duubay spectrometer-midab sagaal ah.Dhaqdhaqaaqa xoojinta ee sagaal-midab (C1-C9) fluorescence inta lagu jiro electrophoresis, taas oo ah, electrophoregram sagaal midab ah oo ka mid ah xidid kasta, ayaa lagu muujiyay sawirka 7a.Electrophoregram sagaal midab u dhigma ayaa laga helay Cap1-Cap4.Sida lagu muujiyey fallaadhaha Cap1 ee Jaantuska 7a, siddeedda ugu sarreysa ee sagaal kasta oo elektrophoregram ah ayaa muujinaya hal qiiqa fluorescence ee Dye1-Dye8, siday u kala horreeyaan.
Qiyaasta isku mar ah ee siddeed midabaynta iyadoo la adeegsanayo spectrometer afar-kaabaysan oo sagaal midab leh.(a) Sagaal-midab (C1-C9) electrophoregram ee xidid kasta.Siddeedda sare ee fallaadhaha Cap1 waxay muujinayaan sii daayo fluorescence ee siddeed midab (Dye1-Dye8).Midabada fallaadhaha waxay u dhigmaan midabada (b) iyo (c).(b) Muuqaalka Fluorescence ee siddeed midab (Dye1-Dye8) halkii xididba.c Electropherograms siddeed midab leh (Dye1-Dye8) halkii xididba.Meelaha ugu sarreeya ee jajabyada DNA-da ee lagu calaamadiyay Dye7 waxaa lagu muujiyey fallaadho, iyo dhererkooda Cap4 waa la tilmaamay.
Qaybinta xoojinta C1-C9 ee siddeedda ugu sarreysa ayaa lagu muujiyay berdihii.7b, siday u kala horreeyaan.Sababtoo ah C1-C9 iyo Dye1-Dye8 labaduba waxay u kala horreeyaan mawjada dhererka, siddeedda qaybinta ee Jaantuska. 7b waxay muujinayaan muraayadda fluorescence ee Dye1-Dye8 oo isku xigta bidix ilaa midig.Daraasaddan, Dye1, Dye2, Dye3, Dye4, Dye5, Dye6, Dye7, iyo Dye8 waxay ka soo muuqdaan magenta, violet, buluug, cyan, cagaar, jaalle, orange, iyo casaan, siday u kala horreeyaan.Ogow in midabada fallaadhaha ee sawirka 7a ay u dhigmaan midabada dheeha ee sawirka 7b.Xoojinta fluorescence-ka C1-C9 ee spectrum kasta ee Jaantuska 7b waa la caadiyeeyay si ay wadartoodu u siman tahay mid.Siddeed muuqaal oo u dhigma spectra fluorescence ayaa laga helay Cap1-Cap4.Qofku wuxuu si cad u eegi karaa isku dhafka muuqalka ee fluorescence ee u dhexeeya dheeha 1-dheeha 8.
Sida ku cad Jaantuska 7c, xidid kasta, sagaalka midab ee elektrophoregram ee Jaantuska 7a waxa loo beddelay korantada sided-midab leh oo lagu falanqeeyay qaybo badan oo ku salaysan siddeedda fluorescence spectra ee Jaantuska 7b (fiiri agabka dheeriga ah wixii faahfaahin ah).Maaddaama isku-ku-dhowrka muraayadda ee Jaantuska 7a aan lagu muujin Jaantuska 7c,Dye1-Dye8 waa la aqoonsan karaa oo si gaar ah ayaa loo qiyaasi karaa wakhti kasta, xitaa haddii xaddi kala duwan oo Fye1-Dye8 fluoresce ah isku mar.Tan laguma samayn karo ogaanshaha todobada midab ee caadiga ah31, laakiin waxa lagu gaadhi karaa ogaanshaha sagaal midab leh.Sida ay muujinayaan fallaadhaha Cap1 ee Jaantuska 7c, kaliya qiiqa qiiqa sii daaya Dye3 (buluug), Dye8 (casaan), Dye5 (cagaar), Dye4 (cyan), Dye2 (guduud), Dye1 (magenta), iyo Dye6 (Jaalle). ) lagu arkay siday u kala horreeyaan.Siideynta fluorescent ee dheeha 7 (orange), marka lagu daro halka ugu sarreysa ee ay tilmaantay fallaadha liimiga, dhowr meelood oo kale ayaa la arkay.Natiijadan waxaa sabab u ah xaqiiqda ah in shaybaaradu ka kooban yihiin halbeegyada cabbirka, Dye7 lagu calaamadiyay jajab DNA oo dhererkoodu kala duwan yahay.Sida ku cad Jaantuska 7c, ee Cap4 dhererkani waa 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214 iyo 220 dhererka salka.
Astaamaha ugu muhiimsan ee spectrometer-ka sagaal-midabka ah, oo la sameeyay iyadoo la adeegsanayo jaantuska muraayadaha laba-lakabka ah ee dichroic, waa cabbir yar iyo naqshad fudud.Tan iyo soo diyaarinta muraayadaha decachromatic gudaha adabtarada lagu muujiyey berdaha.3c oo si toos ah loogu dhejiyay guddiga dareenka sawirka (eeg sawirka S1 iyo S2), spectrometer-ka sagaal-midabka leh wuxuu leeyahay cabbir la mid ah kan adabtarada, ie 54 × 58 × 8.5 mm.( dhumucdiisuna) .Cabbirkan aadka u yar waa laba ilaa saddex amar oo cabbir ka yar cabbirrada caadiga ah ee isticmaala sharoobada ama prisms.Intaa waxaa dheer, maadaama spectrometer-ka midab-takoorka ah ee sagaal-midabka ah loo habeeyey si iftiinku u garaaco dusha sare ee dareenka sawirka si siman, meel bannaan ayaa si fudud loogu qoondeyn karaa spectrometer-ka sagaal-midabka ee nidaamyada sida microscopes, cytometers socodka, ama falanqeeyayaasha.Falanqeeyaha korantada korantada ee xididdada xididka ee xitaa yaraynta nidaamka.Isla mar ahaantaana, cabbirka toban muraayadaha dichroic iyo filtarrada bandpass ee loo isticmaalo spectrometer-ka sagaal-midabku waa kaliya 10 × 1.9 × 0.5 mm ama 15 × 1.9 × 0.5 mm.Sidaa darteed, in ka badan 100 muraayadaha yar yar ee dichroic iyo filtarrada bandpass, siday u kala horreeyaan, ayaa laga gooyn karaa muraayadda dichroic iyo filter filter 60 mm2, siday u kala horreeyaan.Sidaa darteed, muraayado kala duwan oo muraayadaha decachromatic ah ayaa lagu soo saari karaa qiimo jaban.
Muuqaal kale oo ka mid ah spectrometer-ka sagaal-midabka leh ayaa ah sifooyin muuqaal ah oo aad u fiican.Gaar ahaan, waxay u ogolaataa helitaanka sawirada muuqaalka sawirada sawirada, taas oo ah, helitaanka isku mar ee sawirada leh macluumaadka muuqaalka.Sawir kasta, spectrum joogto ah ayaa la helay iyada oo dhererkeedu yahay 520 ilaa 700 nm iyo xallinta 20 nm.Si kale haddii loo dhigo, sagaal midab oo iftiin ah ayaa lagu ogaanayaa sawir kasta, tusaale ahaan sagaal 20 nm oo isku mid ah oo u qaybinaya dhererka dhererka 520 ilaa 700 nm.Adiga oo bedelaya sifooyinka muuqaalka muraayadda dichroic iyo filter-ka bandpass, dhererka dhererka dhererka ee sagaalka qaybood iyo ballaca koox kasta waa la hagaajin karaa.Sagaal ogaanshaha midabka looma isticmaali karo kaliya cabirrada fluorescence oo leh sawir muuqaal ah (sida lagu sharaxay warbixintan), laakiin sidoo kale codsiyo kale oo badan oo caan ah ayaa loo isticmaali karaa sawir muuqaal ah.Inkasta oo sawirka hyperspectral uu ogaan karo boqollaal midab, waxaa la ogaaday in xitaa hoos u dhac weyn oo ku yimid tirada midabada la ogaan karo, walxo badan oo ku yaal goobta aragtida ayaa lagu aqoonsan karaa saxsanaan ku filan codsiyo badan38,39,40.Sababtoo ah xallinta meel bannaan, xallinta muuqaalka, iyo xallinta ku-meel-gaadhka ah waxay leeyihiin isbeddel xagga sawirka muuqaalka ah, yaraynta tirada midabada waxay hagaajin kartaa xallinta goobta iyo xallinta ku meel gaarka ah.Waxa kale oo ay isticmaali kartaa spectrometers fudud sida kan lagu sameeyay daraasaddan oo ay sii yarayso tirada xisaabinta.
Daraasaddan, siddeed midabyo ayaa isku mar lagu qiyaasey iyadoo la eegayo kala soocida muuqaalkooda ee fluorescence ee isku xiran iyadoo lagu salaynayo ogaanshaha sagaal midab.Ilaa sagaal midabyo ayaa isku mar lagu qiyaasi karaa, iyaga oo ku wada nool waqti iyo meel.Faa'iidada gaarka ah ee spectrometer-ka sagaal-midabka leh waa qulqulka iftiinka sare iyo daloolka weyn (1 × 7 mm).Qaabka muraayadda decane waxa ay leedahay gudbinta ugu badan ee 92% ee iftiinka daloolka daloolka mid kasta oo ka mid ah safafka dhererka mawjadda.Waxtarka isticmaalka iftiinka dhacdada ee kala duwanaanshaha hirarka dhererka 520 ilaa 700 nm waa ku dhawaad 100%.Mawjadaha dhererka ballaaran ee noocaas ah, ma jiro sharoobada kala-baxa ah oo ku siin kara waxtarka sare ee isticmaalka.Xataa haddii hufnaanta kala duwanaanshiyaha shabaqlaha diffraction uu dhaafo 90% mawjadda dhererka cayiman, sida farqiga u dhexeeya hirarka dhererka iyo hirarka gaar ah kordho, waxtarka diffraction ee mawjada kale hoos u dhaco41.Balaadhka daloolka ee ku toosan jihada diyaaradda ee Jaantuska 2c waxa laga kordhin karaa 7 mm ilaa ballaca dareeraha sawirka, sida xaalada shidma sawirka ee daraasaddan loo adeegsaday, iyada oo wax yar wax laga beddelayo isku xidhka qalabaynta.
spectrometer-ka sagaal-midabka ah waxaa loo isticmaali karaa ma aha oo kaliya electrophoresis capillary, sida lagu muujiyay daraasaddan, laakiin sidoo kale ujeedooyin kala duwan.Tusaale ahaan, sida ku cad shaxanka hoose, spectrometer-midab leh oo sagaal ah ayaa lagu dabaqi karaa mikroskoob-ka fluorescence.Diyaarada muunada waxa lagu muujiyaa dareenka sawirka ee spectrometer-ka midabka sagaalka ah iyada oo loo marayo ujeedo 10x ah.Fogaanta indhaha ee u dhaxaysa muraayada ujeedada iyo dareenka sawirku waa 200 mm, halka fogaanta indhaha ee u dhaxaysa dusha dhacdada ee spectrometer-ka midabka ah iyo sawirku waa 12 mm oo keliya.Sidaa darteed, sawirka ayaa la gooyay qiyaastii xajmiga daloolka (1 × 7 mm) ee diyaaradda dhacdada waxaana loo qaybiyay sagaal sawir oo midab leh.Taasi waa, sawir muuqaal ah oo sawir sagaal midab leh ayaa lagu qaadi karaa aagga 0.1 × 0.7 mm ee diyaaradda muunada.Intaa waxaa dheer, waxaa suurtogal ah in la helo sawir muuqaal ah oo sagaal midab leh oo ah aag weyn oo ku yaal diyaaradda muunada iyadoo la sawirayo muunada marka la eego ujeedada jihada toosan ee sawirka 2c.
Qaybaha isku dubaridka muraayadda decachromatic, kuwaas oo kala ah M1-M9 iyo BP, waxaa si gaar ah u sameeyay Asahi Spectra Co., Ltd. iyadoo la isticmaalayo hababka roobabka caadiga ah.Qalabka dielectric Multilayer ayaa si gaar ah loogu dabaqay toban taarikada quartz 60 × 60 mm cabbirka iyo 0.5 mm dhumucdiisuna, buuxinta shuruudaha soo socda: M1: IA = 45 °, R ≥ 90% at 520-590 nm, Tave ≥ 90% at 610- 610 nm.700 nm, M2: IA = 45 °, R ≥ 90% at 520-530 nm, Tave ≥ 90% at 550-600 nm, M3: IA = 45 °, R ≥ 90% at 540-550 nm, Tave 90 ≥ % at 570-600 nm, M4: IA = 45°, R ≥ 90% at 560-570 nm, Tave ≥ 90% at 590-600 nm, M5: IA = 45°, R ≥ 98% at 580-600 nm , R ≥ 98% at 680-700 nm, M6: IA = 45 °, Tave ≥ 90% at 600-610 nm, R ≥ 90% at 630-700 nm, M7: IA = 45 °, R ≥ 90% 620-630 nm, Taw ≥ 90% at 650-700 nm, M8: IA = 45°, R ≥ 90% at 640-650 nm, Taw ≥ 90% at 670-700 nm, M9: IA = 45° 90% at 650-670 nm, Tave ≥ 90% at 690-700 nm, BP: IA = 0 °, T ≤ 0.01% at 505 nm, Tave ≥ 95% at 530-690 nm at 530 nm at -690 nm iyo T ≤ 1% at 725-750 nm, halkaas oo IA, T, Tave, iyo R yihiin xagasha dhacdooyinka, gudbinta, celceliska gudbinta, iyo milicsiga iftiinka aan sinnayn.
Iftiin cad (C0) oo leh dherer dhererkiisu yahay 400-750 nm oo ay sii daysay isha iftiinka LED (AS 3000, AS ONE CORPORATION) ayaa isku dhacay oo si toos ah u dhacday DP ee isku dubaridka muraayadaha dichroic.Muuqaalka iftiinka cad ee LED-yada waxa lagu muujiyay sawirka Dheeraadka ah S3.Dhig taangiga akril (cabirkiisu yahay 150 × 150 × 30 mm) si toos ah uga soo hor jeedda muraayadda muraayadda, kasoo horjeedka PSU.Qiiqa ka dhashay markii baraf qallalan lagu dhex shubay biyaha ayaa markaas lagu shubay haan akrilik ah si loo eego sagaalka midab ee C1-C9 ee durdurrada kala qaybsan ee ka soo baxaya isku dubaridka muraayadaha decachromatic.
Haddii kale, iftiinka cad ee isku-dhafka ah (C0) ayaa la dhex maraa filter ka hor inta aan la gelin DP.Shaandheeyayaashu markii hore waxay ahaayeen filtarrada cufnaanta dhexdhexaadka ah oo leh cufnaanta indhaha ee 0.6.Ka dib isticmaal shaandhada matoorka leh (FW212C, FW212C, Thorlabs).Ugu dambeyntii, shaandhada ND dib u shid.Xadhkaha xadhkaha xadhkaha goosha ee sagaalka bandpass waxay u dhigmaan C9, C8, C7, C6, C5, C4, C3, C2 iyo C1, siday u kala horreeyaan.Unug quartz ah oo leh cabbir gudaha ah oo ah 40 (dhererka indhaha) x 42.5 (dherer) x 10 mm (ballaadh) ayaa la hor dhigay soo diyaarsan muraayadaha decochromatic, oo ka soo horjeeda BP.Qiiqa ayaa markaa lagu quudiyaa tuubo la gelinayo unugga quartz-ka si loo ilaaliyo xoogga qiiqa ee unugga quartz-ka si loo sawiro sagaalka midab ee C1-C9 ee durdurrada kala qaybsan ee ka soo baxa muraayadda muraayadda decachromatic.
Fiidiyow ka mid ah qulqulka iftiinka kala qaybsan ee sagaal midab leh ee ka soo baxaya muraayado badan oo muraayado ah ayaa lagu qabtay qaab waqti-dhaaf ah iPhone XS.Sawirada goobta ka qaad 1fps oo samee sawirada si aad u abuurto fiidyow 30fps (fiidiyow ikhtiyaari ah 1) ama 24fps (fidiyayaal ikhtiyaari ah 2 iyo 3).
Dhig saxan bir ah oo qaro weyn oo ah 50 µm (oo leh afar dalool oo dhexroor ah 50 µm oo u dhexeeya 1 mm) saxanka faafinta.Iftiin leh mawjadda dhererka 400-750 nm ayaa lagu shubay saxanka faafiyaha, oo laga helo iftiinka ka soo baxa nalka halogen iyada oo loo marayo filtar gudbin gaaban oo leh mawjadda gooyay ee 700 nm.Muuqaalka iftiinka waxa lagu muujiyay sawirka Dheeraadka ah S4.Haddii kale, iftiinku wuxuu sidoo kale maraa mid ka mid ah 10 nm filtarrada bandpass ee xuddun u ah 530, 550, 570, 590, 610, 630, 650, 670 iyo 690 nm wuxuuna ku dhuftaa saxanka faafiyaha.Natiijo ahaan, afar dhibcood oo shucaac ah oo leh dhexroor φ50 μm iyo hirarka hirarka kala duwan ayaa lagu sameeyay saxan bir ah oo ka soo horjeeda saxanka faafiyaha.
Qalab afar geesood ah oo leh afar lenses ayaa lagu rakibay spectrometer sagaal midab leh sida ku cad jaantusyada 1 iyo 2. C1 iyo C2.Afarta indhoolayaasha iyo afarta lenses waxay la mid ahaayeen daraasadihii hore31,34.Laydhka laysarka oo leh mowjadda dhererka 505 nm iyo awoodda 15 mW ayaa si isku mar ah iyo si siman uga soo ifbaxaya dhinaca ilaa dhibcaha qiiqa ee afar xabbo.Foluorescence-ka ka soo baxa barta qiiqa ka sii daayaba waxa isku xidha muraayada u dhiganta waxaana u kala qaybiyaa sagaal durdurro midab leh oo ay isku dubaridaan muraayado decachromatic ah.Natiijadii 36 durdurrada ayaa markaa si toos ah loogu duray sawirka sawirka CMOS (C11440-52U, Hamamatsu Photonics K·K.), Sawirradoodana waa la duubay.
ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit (Noolaha la dabaqay), 4 µl GeneScan™ 600 LIZ™ dheeha ayaa lagu qasay xidid kasta iyadoo la isku daray 1 µl PowerPlex® 6C Matrix Standard (Promega Corporation), 1 µl heerka cabbirka isku dhafka ah.v2.0 (Thermo Fisher Scientific) iyo 14 µl oo biyo ah.Halbeegga Matrix PowerPlex® 6C wuxuu ka kooban yahay lix jajab oo DNA ah oo lagu calaamadeeyay lix midab: FL-6C, JOE-6C, TMR-6C, CXR-6C, TOM-6C, iyo WEN, siday u kala horreeyaan mawjadaha dhererka ugu badan.Dhererka saldhigga ee jajabyada DNA-da lama shaacin, laakiin isku xigxiga dhererka saldhigga ee jajabyada DNA-da ee ku calaamadsan WEN, CXR-6C, TMR-6C, JOE-6C, FL-6C iyo TOM-6C waa la yaqaan.Isku darka ku jira ABI PRISM® BigDye® Primer Cycle Sequencing Ready Reaction Kit wuxuu ka kooban yahay jajab DNA ah oo lagu calaamadeeyay dheeha dR6G.Dhererka saldhigyada jajabyada DNA-da sidoo kale lama shaacin.GeneScan™ 600 LIZ™ Heerka Dheedhiga v2.0 waxa ku jira 36 jajab oo DNA ah oo LIZ ku suntan.Dhererka saldhigyada jajabyada DNA-da waa 20, 40, 60, 80, 100, 114, 120, 140, 160, 180, 200, 214, 220, 240, 250, 260, 280, 30,30, 30,300 360, 380, 400, 414, 420, 440, 460, 480, 500, 514, 520, 540, 560, 580 iyo 600 saldhig.Tijaabooyinku waxa laga saaray 94°C 3 daqiiqo, ka dibna baraf lagu qaboojiyey 5 daqiiqo.Tijaabooyin ayaa lagu duray xidid kasta oo ah 26 V/cm 9 s waxaana lagu kala soocay xidid kasta oo ay ka buuxaan POP-7 ™ polymer solution (Thermo Fisher Scientific) oo leh dherer wax ku ool ah oo ah 36 cm iyo danab 181 V / cm ah iyo xagal 60°.KA.
Dhammaan xogta la helay ama la falanqeeyay intii ay socotay daraasaddan waxay ku jirtaa maqaalkan la daabacay iyo macluumaadkiisa dheeraadka ah.Xogta kale ee la xidhiidha daraasaddan ayaa laga heli karaa qorayaasha kala duwan marka la eego codsi macquul ah.
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Waqtiga boostada: Jan-10-2023