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Ubonisa ikharawusi yezilayidi ezithathu ngaxeshanye.Sebenzisa amaqhosha angaphambili nalandelayo ukutyhutyha izilayidi ezithathu ngexesha, okanye sebenzisa amaqhosha esilayidi ekupheleni ukuya kwizilayidi ezithathu ngexesha.
I-Confocal laser endoscopy yindlela entsha ye-real-time optical biopsy.Imifanekiso ye-Fluorescent yomgangatho we-histological inokufumaneka ngokukhawuleza kwi-epithelium yezitho ezingenanto.Okwangoku, ukuskena kwenziwa kufutshane nezixhobo ezisekwe kwiprobe ezidla ngokusetyenziswa kuqheliselo lweklinikhi, kunye nokuguquguquka okulinganiselweyo kulawulo lokugxila.Sibonisa ukusetyenziswa kwe-parametric resonant scanner efakwe kwi-distal end ye-endoscope ukwenza i-high-speed lateral deflection.Kuye kwakrolwa umngxuma kumbindi wesibonisi ukusonga indlela yokukhanya.Olu lwakhiwo lunciphisa ubungakanani besixhobo ukuya kwi-2.4 mm ububanzi kunye ne-10 mm ubude, ukuvumela ukuba kudluliselwe phambili kumjelo osebenzayo wee-endoscopes zonyango eziqhelekileyo.I-compact lens ibonelela ngezigqibo ezisecaleni kunye ne-axial ze-1.1 kunye ne-13.6 µm, ngokulandelelanayo.Umgama wokusebenza we-0 µm kunye nebala lokujonga i-250 µm × 250 µm zifunyenwe kumazinga esakhelo ukuya kuma-20 Hz.Uchulumanco kwi-488 nm iyayivuyisa i-fluorescein, idayi evunyiweyo ye-FDA yokuthelekisa izicubu eziphezulu.Ii-Endoscopes ziphinde zacutshungulwa kwimijikelo eli-18 ngaphandle kokusilela kusetyenziswa iindlela zokuvala inzala ezivunyiweyo ngokwasekliniki.Imifanekiso ye-Fluorescent ifunyenwe kwi-colonic mucosa eqhelekileyo, i-tubular adenomas, i-hyperplastic polyps, i-ulcerative colitis, kunye ne-Crohn's colitis ngexesha le-colonoscopy yesiqhelo.Iiseli ezingatshatanga ziyakwazi ukuchongwa, kuquka i-colonocytes, iiseli ze-goblet, kunye neeseli ezivuthayo.Iimpawu ze-Mucosal ezifana nezakhiwo ze-crypt, i-crypt cavities, kunye ne-lamina propria zinokuhlukaniswa.Isixhobo sinokusetyenziswa njengesincedisi kwi-endoscopy yesiqhelo.
I-confocal laser endoscopy yindlela yokucinga yenoveli ephuhliswayo ukuze isetyenziswe njengesiqhelo kwi-endoscopy1,2,3.Ezi zixhobo eziguquguqukayo, ezixhunyiwe kwi-fiber-optic zingasetyenziselwa ukufumanisa izifo kwiiseli ze-epithelial ezidibanisa izitho ezingenanto, ezifana nekholoni.Lo maleko obhityileyo wenyama uyasebenza kakhulu kwaye ungumthombo weenkqubo ezininzi zezifo ezinjengomhlaza, usulelo kunye nokudumba.I-Endoscopy inokufikelela kwisisombululo se-subcellular, inikezela ngexesha langempela, umgangatho osondeleyo we-histological kwimifanekiso ye-vivo ukunceda oogqirha ukuba benze izigqibo zekliniki.I-biopsy yezicubu zomzimba inomngcipheko wokuphuma kwegazi kunye nokubhoboza.Imizekelo emininzi kakhulu okanye embalwa kakhulu ye-biopsy isoloko iqokelelwa.Isampulu nganye esusiweyo yonyusa iindleko zotyando.Kuthatha iintsuku ezininzi ukuba isampulu ivavanywe ngugqirha wezifo.Ngethuba leentsuku zokulinda iziphumo ze-pathology, izigulane zihlala ziba nexhala.Ngokwahlukileyo, ezinye iindlela zokucinga zekliniki ezifana ne-MRI, CT, PET, SPECT, kunye ne-ultrasound ayinaso isisombululo sendawo kunye nesantya sexeshana esifunekayo ukujonga iinkqubo ze-epithelial kwi-vivo kunye nexesha langempela, isisombululo se-subcellular.
Isixhobo esisekwe kwiprobe (Cellvizio) okwangoku sisetyenziswa ngokuqhelekileyo kwiikliniki ukwenza "i-optical biopsy".Uyilo lusekwe kwi-spatially coherent fiber optic bundle4 eqokelela kwaye ithumele imifanekiso yefluorescent.I-fiber core enye isebenza "njengomngxuma" wokucoca indawo yokukhanya okungagxininisi kwisisombululo se-subcellular.Ukuskena kwenziwa ngokusondeleyo kusetyenziswa igalvanometer enkulu, enkulu.Esi sibonelelo sinciphisa ukukwazi kwesixhobo sokulawula ukugxila.Ukubekwa ngokufanelekileyo kwe-epithelial carcinoma yokuqala kufuna ukubonwa ngaphantsi komphezulu we-tissue ukuvavanya ukuhlasela kunye nokumisela unyango olufanelekileyo.I-Fluorescein, i-arhente echaseneyo evunywe yi-FDA, ilawulwa nge-intravenously ukugqamisa iimpawu zesakhiwo se-epithelium. Ezi ze-endomicroscopes zinemilinganiselo <2.4 mm ububanzi, kwaye zinokugqithiselwa phambili ngokulula ngejelo le-biopsy yee-endoscopes zonyango eziqhelekileyo. Ezi ze-endomicroscopes zinemilinganiselo <2.4 mm ububanzi, kwaye zinokugqithiselwa phambili ngokulula ngejelo le-biopsy yee-endoscopes zonyango eziqhelekileyo. Эти эндомикроскопы имеют размеры Ezi zi-endomicroscopes zi-<2.4 mm ububanzi kwaye zinokugqithiswa lula ngejelo le-biopsy yee-endoscopes zonyango eziqhelekileyo.Ezi borescopes zingaphantsi kwe-2.4 mm ububanzi kwaye zidlula ngokulula kumjelo we-biopsy yeebhorescope zonyango eziqhelekileyo.Oku kuguquguquka kuvumela uluhlu olubanzi lwezicelo zeklinikhi kwaye luzimeleyo kubenzi be-endoscope.Izifundo ezininzi zeklinikhi zenziwe kusetyenziswa esi sixhobo sokucinga, kubandakanya ukufunyanwa kwangoko komhlaza wommizo, wesisu, wekholoni kunye nowomlomo womlomo.Iiprothokholi zomfanekiso zenziwe kwaye ukhuseleko lwenkqubo lusekiwe.
Iinkqubo zeMicroelectromechanical (MEMS) bubuchwephesha obunamandla bokuyila kunye nokuvelisa iindlela ezincinci zokuskena ezisetyenziswa kwisiphelo esikude se-endoscopes.Esi sikhundla (ngokumalunga ne-proximal) sivumela ukuguquguquka okukhulu ekulawuleni indawo yokugxila5,6.Ukongeza kwi-lateral deflection, i-distal mechanism inokwenza i-axial scans, i-post-objective scans, kunye ne-random access scans.Ezi zakhono zenza ukuba uphononongo olubanzi lweeseli ze-epithelial, kubandakanywa umfanekiso onqamlezayo othe nkqo wecandelo7, indawo enkulu yokujonga (FOV)8 ukuskena okungahambi kakuhle, kunye nokusebenza okuphuculweyo kwiingingqi ezichazwe ngumsebenzisi9.I-MEMS isombulula ingxaki enkulu yokupakisha i-injini yokuskena kunye nendawo encinci ekhoyo ekupheleni kwesixhobo.Xa kuthelekiswa neegalvanometers ezinkulu, i-MEMS ibonelela ngokusebenza okuphezulu kwisayizi encinci, isantya esiphezulu, kunye nokusetyenziswa kwamandla aphantsi.Inkqubo elula yokuvelisa inokunyuswa kwimveliso eninzi ngexabiso eliphantsi.Uyilo oluninzi lwe-MEMS luye lwaxelwa ngaphambili10,11,12.Akukho nanye itekhnoloji esele iphuhliswe ngokwaneleyo ukuvumela ukusetyenziswa kweklinikhi okuxhaphakileyo kwexesha lokwenyani kwi-vivo imaging ngokusebenzisa umjelo osebenzayo we-endoscope yezonyango.Apha, sijonge ukubonisa ukusetyenziswa kweskena se-MEMS ekupheleni kwe-endoscope kwi-vivo yokufumana umfanekiso womntu ngexesha le-endoscopy yesiqhelo yeklinikhi.
Isixhobo se-fiber optic saphuhliswa kusetyenziswa iskena se-MEMS ekupheleni kwe-distal ukuqokelela real-time kwi-vivo fluorescent imifanekiso eneempawu ezifanayo ze-histological.I-fiber yemodi enye (i-SMF) ifakwe kwi-tube ye-polymer eguquguqukayo kwaye iyavuya kwi-λex = 488 nm.Olu lungelelwaniso lufutshane ubude bencam ye-distal kwaye ivumela ukuba idluliselwe phambili ngejelo elisebenzayo leendoscopes zonyango eziqhelekileyo.Sebenzisa incam ukubeka embindini i-optic.Ezi lens ziyilelwe ukufikelela phantse diffractive axial isisombululo nge aperture yamanani (NA) = 0.41 kunye nomgama wokusebenza = 0 µm13.Iishim ezichanekileyo zenziwa ukulungelelanisa ngokuchanekileyo i-optics 14. Iskena sipakishwe kwi-endoscope ene-distal tip eqinile 2.4 mm ububanzi kunye ne-10 mm ubude (Fig. 1a).Le milinganiselo ivumela ukuba isetyenziswe kwikliniki njengento yokuncedisa ngexesha le-endoscopy (Umfanekiso 1b).Amandla aphezulu esiganeko se-laser kwi-tissue yayiyi-2 mW.
I-Confocal laser endoscopy (CLE) kunye neeskena ze-MEMS.Ifoto ebonisa (a) isixhobo esipakishweyo esinencam eqinileyo yomlinganiselo we-2.4 mm ubukhulu kunye ne-10 mm ubude kunye (b) nendlela ethe ngqo kumjelo osebenzayo we-endoscope yonyango eqhelekileyo (Olympus CF-HQ190L).(c) Umbono ongaphambili weskena obonisa isibonisi esinomkhala osembindini wama-50 µm apho kudlula umqa wovuselelo.Iskena sixhonywe kwigimbal eqhutywa liqela le quadrature comb drive drives.Ubuninzi be-resonant yesixhobo buchongwa ngubungakanani bentwasahlobo ye-torsion.(d) Umbono osecaleni weskena esibonisa iskena esixhonywe kwisitendi esineengcingo eziqhagamshelwe kwii-ankile ze-electrode ezibonelela ngeendawo zoqhagamshelo lweempawu zokuqhuba kunye nombane.
Umatshini wokuskena uquka i-reflector enyuswe kwigimbal eqhutywa liqela le-actuator ye-quadrature eqhutywa yikama ukuphambukisa umqadi ecaleni (inqwelomoya ye-XY) kwipatheni ye-Lissajous (Fig. 1c).Umngxuma oyi-50 µm ububanzi waqanjwa embindini apho umqa wovuselelo ugqithe kuwo.Iskena siqhutywa kwi-resonant frequency yoyilo, enokulungiswa ngokutshintsha imilinganiselo yomthombo we-torsion.Ii-anchors ze-Electrode zazibhalwe kwi-periphery yesixhobo ukubonelela ngamanqaku okudibanisa amandla kunye neempawu zokulawula (Umfanekiso 1d).
Inkqubo yokucinga ifakwe kwinqwelo ephathekayo enokuthi ifakwe kwigumbi lokusebenza.Igraphical interface yomsebenzisi iyilelwe ukuxhasa abasebenzisi abanolwazi oluncinci lobuchwephesha, abafana noogqirha nabongikazi.Jonga ngesandla i-scanner drive frequency, beamform mode, kunye nomfanekiso weFOV.
Ubude obupheleleyo be-endoscope bumalunga ne-4m ukuvumela ukugqithwa okupheleleyo kwezixhobo kumjelo osebenzayo we-endoscope yonyango eqhelekileyo (1.68m), kunye nobude obongezelelweyo bokuhamba.Kwisiphelo esisondeleyo se-endoscope, i-SMF kunye neengcingo ziphela kwizixhumi ezidibanisa kwi-fiber optic kunye neengcingo ze-wire station station.Ufakelo luqulethe i-laser, iyunithi yokucoca, i-amplifier high-voltage kunye ne-photomultiplier detector (PMT).Iamplifier inika amandla kunye neempawu zokuqhuba kwiskena.Iyunithi yokucoca i-optical filter idibanisa i-laser excitation kwi-SMF kwaye igqithise i-fluorescence kwi-PMT.
Ii-Endoscopes ziphinda zenziwe kwakhona emva kwenkqubo nganye yekliniki kusetyenziswa inkqubo yokuvala inzala ye-STERRAD kwaye inokumelana ukuya kuthi ga kwimijikelo eli-18 ngaphandle kokusilela.Kwisisombululo se-OPA, akukho zimpawu zomonakalo zabonwa emva kwemijikelo ye-disinfection engaphezu kwe-10.Iziphumo ze-OPA ziye zagqwesa ii-STERRAD's, zicebisa ukuba ubomi be-endoscopes bunokwandiswa ngokubulala iintsholongwane okukwinqanaba eliphezulu kunokuphinda kubulawe inzala.
Isisombululo somfanekiso sinqunywe ukusuka kumsebenzi wokusasazwa kwendawo kusetyenziswa amaso e-fluorescent kunye nobubanzi be-0.1 μm.Ukufumana isisombululo secala kunye ne-axial, ububanzi obugcweleyo kwisiqingatha esiphezulu (FWHM) se-1.1 kunye ne-13.6 µm, ngokulandelelanayo, yalinganiswa (Umfanekiso 2a, b).
Iinketho zomfanekiso.I-lateral (a) kunye ne-axial (b) isisombululo se-optics egxininisekile ibonakaliswe ngumsebenzi wokusasazeka kwenqaku (PSF) elilinganiswe kusetyenziswa ii-microspheres ze-fluorescent ezinobubanzi be-0.1 μm.Ububanzi obulinganisiweyo obupheleleyo kwisiqingatha sobuninzi (FWHM) yayiyi-1.1 kunye ne-13.6 µm, ngokulandelelanayo.Isingeniso: Iimbono ezandisiweyo zemicrosphere enye kwi-transverse (XY) kunye ne-axial (XZ) izikhombisi ziyaboniswa.(c) Umfanekiso we-Fluorescent ofunyenwe kwi-standard (USAF 1951) i-target strip (i-oval ebomvu) ebonisa ukuba amaqela 7-6 anokusombulula ngokucacileyo.(d) Umfanekiso we-10 µm idayamitha esasaziweyo yemicrospheres efluorescent ebonisa indawo yomfanekiso we-250 µm×250 µm.Ii-PSF ku-(a, b) zakhiwe kusetyenziswa iMATLAB R2019a (https://www.mathworks.com/).(c, d) Imifanekiso yeFluorescent yaqokelelwa kusetyenziswa iLabVIEW 2021 (https://www.ni.com/).
Imifanekiso ye-Fluorescent evela kwiilensi zesisombululo esiqhelekileyo ihlukanisa ngokucacileyo isethi yeekholomu kumaqela 7-6, egcina isisombululo esiphezulu secala (Umfanekiso 2c).Indawo yokujonga (FOV) ye-250 µm × 250 µm yamiselwa kwimifanekiso ye-10 µm yamaso e-fluorescent edayamitha esasazwe kumaqhina (Fig. 2d).
Indlela ezenzekelayo yokulawula inzuzo ye-PMT kunye nokulungiswa kwesigaba kuphunyezwa kwinkqubo yokucinga yeklinikhi ukunciphisa izinto ezishukumayo ezivela kwi-endoscopes, i-colon peristalsis, kunye nokuphefumla kwesigulane.Ukwakhiwa kwakhona kwemifanekiso kunye ne-algorithms yokucubungula ichazwe ngaphambili14,15.Inzuzo ye-PMT ilawulwa ngumlawuli we-proportional-integral (PI) ukunqanda ukuqina kokuqina16.Inkqubo ifunda ubukhulu bepixel intensity kwisakhelo ngasinye, ibala uthelekiso kunye neempendulo ezidityanisiweyo, kwaye imisela amaxabiso okuzuza kwe-PMT ukuqinisekisa ukuba ubunzulu bepixel bungaphakathi koluhlu oluvumelekileyo.
Ngexesha lokucinga nge-vivo, ukungafani kwesigaba phakathi kwentshukumo yeskena kunye nophawu lolawulo kunokubangela ukufiphala komfanekiso.Ezo ziphumo zinokwenzeka ngenxa yotshintsho kwiqondo lobushushu lesixhobo ngaphakathi emzimbeni womntu.Imifanekiso yokukhanya emhlophe ibonise ukuba i-endoscope yayinxibelelana ne-colonic mucosa eqhelekileyo kwi-vivo (Umfanekiso 3a).Ukufiphaza kweepikseli ezingalungelelanisiyo kunokubonwa kwimifanekiso ekrwada ye-colonic mucosa eqhelekileyo (Umfanekiso 3b).Emva kokonyango kunye nesigaba esifanelekileyo kunye nokulungelelaniswa okungafaniyo, iimpawu ze-subcellular ze-mucosa zingahlukaniswa (umzobo 3c).Ngolwazi olongezelelweyo, imifanekiso ye-confocal eluhlaza kunye nemifanekiso yexesha langempela iboniswe kwi-Fig.
Ukulungiswa komfanekiso.(a) Umfanekiso we-engile ebanzi we-endoskophu ebonisa i-endoskophu (E) ebekwe kunxibelelwano lwe-colonic mucosa eqhelekileyo (N) ukuqokelela kwi-vivo imifanekiso yefluorescent emva kolawulo lwe-fluorescein.bNgeenjongo zokubonisa, utshintsho lwesigaba esikhulu lusetyenziswa kumfanekiso wokuqala.(c) Emva kokulungiswa kwesigaba se-post-processing, iinkcukacha ze-mucosal zinokuvavanywa, kubandakanywa izakhiwo ze-crypt (iintolo), kunye ne-lumen ephakathi (l) ejikelezwe yi-lamina propria (lp).Iiseli ezingatshatanga ziyakwazi ukwahlula, kuquka ikholonocytes (c), iiseli zegoblet (g), kunye neeseli ezivuthayo (iintolo).Jonga ividiyo eyongezelelweyo 1. (b, c) Imifanekiso eyenziwe kusetyenziswa iLabVIEW ka-2021.
Imifanekiso ye-confocal fluorescence ifunyenwe kwi-vivo kwizifo ezininzi zekholoni ukubonisa ukusetyenziswa okubanzi kweklinikhi kwesixhobo.Umboniso obanzi wenziwa kuqala kusetyenziswa ukukhanya okumhlophe ukubona umfinya ongaqhelekanga.Emva koko i-endoscope iqhubela phambili ngejelo elisebenzayo lekholonoscope kwaye idibane ne-mucosa.
I-Wide-field endoscopy, i-confocal endomicroscopy, kunye ne-histology (H & E) imifanekiso iboniswa kwi-colonic neoplasia, kuquka i-adenoma ye-tubular kunye ne-hyperplastic polyp. I-Wide-field endoscopy, i-confocal endomicroscopy, kunye ne-histology (H & E) imifanekiso iboniswa kwi-colonic neoplasia, kuquka i-adenoma ye-tubular kunye ne-hyperplastic polyp. Широкопольная эндоскопия, конфокальная эндомикроскопия kunye ne-гистологические (H&E) у и гиперпластический полип. I-endoscopy yekholoni, i-confocal endomicroscopy, kunye ne-histological (H & E) imaging iboniswa kwi-colonic neoplasia, kuquka i-tubular adenoma kunye ne-hyperplastic polyp.显示结肠肿瘤（包括管状腺瘤和增生性息肉）的广角内窻镜检查、共括管状腺瘤和增生性息肉) (H&E) 图像.共 设计 设计设计 脚肠化脚肠化 躰化 增生性息肉 的 的 的 的 的 共 共 共 光 微微全 在 在 圕别 具 和 和 和 结果 结果 结果 Широкопольная эндоскопия, конфокальная микроэндоскопия kunye гистологические (H&E) изображения, показывающие опухоскопия опухоскопия опухоскопия мы и гиперпластические полипы. I-endoscopy ebanzi, i-confocal microendoscopy, kunye nemifanekiso ye-histological (H & E) ebonisa amathumba ekholoni, kuquka i-adenomas ye-tubular kunye ne-hyperplastic polyps.I-adenomas ye-tubular ibonise ukulahlekelwa kwezakhiwo eziqhelekileyo ze-crypt, ukunciphisa ubungakanani beeseli ze-goblet, ukuphazamiseka kwe-crypt lumen, kunye nokuqina kwe-lamina propria (Umfanekiso 4a-c).I-polyps ye-Hyperplastic ibonise i-stellate architecture yee-crypts, iiseli ezimbalwa ze-goblet, i-lumen efana ne-slit ye-crypts, kunye ne-lamellar crypts engaqhelekanga (Umfanekiso we-4d-f).
Umfanekiso wesikhumba esityebileyo se-mucosal kwi-vivo. I-endoscopy yokukhanya okumhlophe emele, i-confocal endomicroscope, kunye ne-histology (H&E) imifanekiso iboniswe (ac) i-adenoma, (df) i-hyperplastic polyp, (gi) i-ulcerative colitis, kunye (jl) ne-Crohn's colitis. I-endoscopy yokukhanya okumhlophe emele, i-confocal endomicroscope, kunye ne-histology (H&E) imifanekiso iboniswe (ac) i-adenoma, (df) i-hyperplastic polyp, (gi) i-ulcerative colitis, kunye (jl) ne-Crohn's colitis. Типичные изображения эндоскопии в белом свете, конфокального эндомикроскопа и гистологии (H&E) показаны для (ac) аденомы, песни, (df) аденомы, (df). нного колита и (jl) колита Крона. I-endoscopy yokukhanya okumhlophe okuqhelekileyo, i-confocal endomicroscope, kunye ne-histology (H&E) imifanekiso iboniswe (ac) adenoma, (df) hyperplastic polyp, (gi) ulcerative colitis, kunye (jl) neCrohn's colitis.显示了(ac) 腺瘤、(df) 增生性息肉、(gi) 溃疡性结肠炎和(jl) 克罗恩结肠炎的代衣全性聚焦内窥镜检查和组织学( H&E) 图像 Ibonisa(ac) 躰真、(df) 增生性息肉、(gi) 苏盖性红肠炎和(jl) 克罗恩红肠炎的体育性白光克罗恩红肠炎的体育性白光内肠肠炎性和电视学( H&E ) umfanekiso. Представлены репрезентативные эндоскопия в белом свете, конфокальная эндоскопия и гистология (ac) аденомы, (df) гипезарплигосплагия (df) l) колита Крона (H&E). I-endoscopy emele ukukhanya okumhlophe, i-endoscopy ye-confocal, kunye ne-histology ye (ac) i-adenoma, (df) i-hyperplastic polyposis, (gi) i-ulcerative colitis, kunye (jl) i-Crohn's colitis (H & E) ibonisiwe.(B) ibonisa umfanekiso we-confocal ofunyenwe kwi-vivo kwi-adenoma ye-tubular (TA) usebenzisa i-endoscope (E).Esi silonda se-precancerous sibonisa ilahleko ye-crypt Architecture eqhelekileyo (utolo), ukuphazamiseka kwe-crypt lumen (l), kunye nokuxinana kwe-crypt lamina propria (lp).I-Colonocytes (c), iiseli zegoblet (g), kunye neeseli ezivuthayo (iintolo) nazo zingachongwa.Smt.Ividiyo eyongezelelweyo 2. (e) ibonisa umfanekiso we-confocal ofunyenwe kwi-hyperplastic polyp (HP) kwi-vivo.Esi silonda sincinci sibonisa i-stellate crypt architecture (utolo), i-crypt lumen (l), kunye ne-lamina propria emile ngendlela engaqhelekanga (lp).IiColonocytes (c), iiseli zegoblet ezininzi (g) kunye neeseli ezivuthayo (iintolo) nazo zinokuchongwa.Smt.Ividiyo eyongezelelweyo 3. (h) ibonisa imifanekiso ye-confocal efunyenwe kwi-ulcerative colitis (UC) kwi-vivo.Le meko yokuvuvukala ibonisa i-architecture ephosakeleyo ye-crypt (utolo) kunye neeseli ze-goblet ezivelele (g).Iintsiba ze-fluorescein (f) zikhutshiwe kwiiseli ze-epithelial, ezibonisa ukunyuka kwe-vascular permeability.Iiseli ezininzi ezivuthayo (iintolo) zibonwa kwi-lamina propria (lp).Smt.Ividiyo eyoNgezelelweyo 4. (k) ibonisa umfanekiso we-confocal ofunyenwe kwi-vivo ukusuka kummandla we-Crohn's colitis (CC).Le meko yokuvuvukala ibonisa i-architecture ephosakeleyo ye-crypt (utolo) kunye neeseli ze-goblet ezivelele (g).Iintsiba ze-fluorescein (f) zikhutshiwe kwiiseli ze-epithelial, ezibonisa ukunyuka kwe-vascular permeability.Iiseli ezininzi ezivuthayo (iintolo) zibonwa kwi-lamina propria (lp).Smt.Ividiyo eyoNgezelelweyo 5. (b, d, h, l) Imifanekiso eyenziwe kusetyenziswa iLabVIEW ka-2021.
Isethi efanayo yemifanekiso yokuvuvukala kwekholoni iboniswa, kubandakanywa i-ulcerative colitis (UC) (Umfanekiso 4g-i) kunye ne-Crohn's colitis (Umfanekiso 4j-l).Impendulo yokuvuvukala kucingelwa ukuba ibonakaliswe ngezakhiwo ezigqwethekileyo ze-crypt ezineeseli ze-goblet eziphumayo.I-Fluorescein ikhutshelwa ngaphandle kweeseli ze-epithelial, ebonisa ukunyuka kwe-vascular permeability.Inani elikhulu leeseli ezivuthayo zingabonwa kwi-lamina propria.
Siye sabonisa ukusetyenziswa kwekliniki ye-flexible fiber-coupled confocal laser endoscope esebenzisa iskena se-MEMS esibekwe kude ukuze ufumane umfanekiso we-vivo.Kwi-resonant frequency, iireyithi zesakhelo ukuya kuthi ga kwi-20 Hz zinokufumaneka kusetyenziswa imowudi yoxinaniso ye-Lissajous ephezulu yokunciphisa izinto ezishukumayo.Umendo obonakalayo uyasongwa ukunika ukwandiswa komqadi kunye nokuvuleka kwamanani okwaneleyo ukufikelela kwisisombululo esisecaleni se-1.1 µm.Imifanekiso yeFluorescent yomgangatho we-histological yafunyanwa ngexesha le-colonoscopy yesiqhelo ye-colonic mucosa eqhelekileyo, i-tubular adenomas, i-hyperplastic polyps, i-ulcerative colitis, kunye ne-Crohn's colitis.Iiseli ezingatshatanga ziyakwazi ukuchongwa, kuquka i-colonocytes, iiseli ze-goblet, kunye neeseli ezivuthayo.Iimpawu ze-Mucosal ezifana nezakhiwo ze-crypt, i-crypt cavities, kunye ne-lamina propria zinokuhlukaniswa.I-hardware echanekileyo i-micro-machined ukuqinisekisa ukulungelelaniswa okuchanekileyo kwezinto ezibonakalayo zomntu ngamnye kunye nomatshini ngaphakathi kwe-2.4mm ububanzi x 10mm isixhobo sobude.Uyilo lwe-optical lunciphisa ubude bencam ye-distal eqinileyo ngokwaneleyo ukuvumela ukudlula ngokuthe ngqo kubukhulu obuqhelekileyo (i-3.2 mm ububanzi) umjelo osebenzayo kwii-endoscopes zonyango.Ngoko ke, kungakhathaliseki ukuba ngumenzi, isixhobo sinokusetyenziswa ngokubanzi oogqirha kwindawo yokuhlala.Uchulumanco lwenziwa e-λex = 488 nm ukonwabisa i-fluorescein, idayi evunyiweyo ye-FDA, ukufumana umahluko ophezulu.Isixhobo saphinda sacutshungulwa ngaphandle kweengxaki kwimijikelo ye-18 kusetyenziswa iindlela zokuvala inzala ezamkelwe eklinikhi.
Ezinye izixhobo ezimbini zoyilo ziye zaqinisekiswa ngokonyango.I-Cellvizio (i-Mauna Kea Technologies) yi-probe-based confocal laser endoscope (pCLE) esebenzisa inyanda yeentambo ze-multimode ezihambelanayo ze-fiber optic ukuqokelela kunye nokuhambisa imifanekiso ye-fluorescence1.Isibuko segalvo esibekwe kwisikhululo esisezantsi senza iskena esisecaleni ekupheleni kweproximal.Amacandelo okukhanya aqokelelwa kwinqwelo-moya ethe tye (XY) enobunzulu obuyi-0 ukuya kuma-70 µm.Iikiti zeMicroprobe zifumaneka kwi-0.91 (19 G inaliti) ukuya kwi-5 mm ububanzi.Isisombululo esisecaleni se-1 ukuya kwi-3.5 µm siphunyeziwe.Imifanekiso iqokelelwe kwireyithi yesakhelo se-9 ukuya kwi-12 Hz kunye nentsimi ene-dimensional yokujonga ukusuka kwi-240 ukuya kwi-600 µm.Iqonga lisetyenziswe eklinikhi kwiindawo ezahlukeneyo ezibandakanya i-bile duct, isinyi, ikholoni, ummizo, imiphunga kunye neepancreas.I-Optiscan Pty Ltd iphuhlise i-endoscope-based confocal laser endoscope (eCLE) enenjini yokuskena eyakhelwe kwi-tube yokufaka (isiphelo se-distal) ye-endoscope yobungcali (EC-3870K, i-Pentax Precision Instruments) 17.Icandelo le-optical liqhutywe ngokusebenzisa i-fiber-mode-mode, kwaye ukuskena kwecala kuqhutywe kusetyenziswa i-cantilever mechanism ngokusebenzisa i-resonant tuning fork.I-activator ye-Shape Memory Memory (Nitinol) isetyenziselwa ukudala i-axial displacement.Ubungakanani obupheleleyo bemodyuli edibeneyo yi-5 mm.Ukugxininisa, i-lens ye-GRIN ene-aperture yamanani ye-NA = 0.6 isetyenziswa.Imifanekiso ethe tyaba ifunyenwe ngezigqibo ezisecaleni kunye ne-axial ye-0.7 kunye ne-7 µm, ngokulandelanayo, kwireyithi yesakhelo se-0.8-1.6 Hz kunye nentsimi yembono ye-500 µm × 500 µm.
Sibonisa isisombululo se-subcellular kwi-vivo fluorescence imaging acquisition evela kumzimba womntu ngokusebenzisa i-endoscope yonyango usebenzisa i-distal end MEMS scanner.I-Fluorescence inikezela ngokungafaniyo komfanekiso ophezulu, kunye ne-ligands ezibophelela kwiithagethi ze-cell surface zingabhalwa nge-fluorophores ukubonelela nge-molecular identity yokuphucula ukuxilongwa kwezifo18.Ezinye iindlela zobuchule bokubona kwi-vivo microendoscopy nazo ziyaphuhliswa. I-OCT isebenzisa ubude obufutshane bokuhambelana ukusuka kumthombo wokukhanya obanzi ukuqokelela imifanekiso kwinqwelomoya ethe nkqo enobunzulu> 1 mm19. I-OCT isebenzisa ubude obufutshane bokuhambelana ukusuka kumthombo wokukhanya obanzi ukuqokelela imifanekiso kwinqwelomoya ethe nkqo enobunzulu> 1 mm19. ОКТ использует короткую длину когерентности широкополосного источника света для сбора изображений в вертикальной плоскости с 196 I-OCT isebenzisa ubude obufutshane bokuhambelana komthombo wokukhanya kwe-broadband ukufumana imifanekiso kwinqwelomoya ethe nkqo nge>1 mm ubunzulu19. OCT 使用宽带光源的短相干长度來收集垂直平面中深度> 1 mm19 的图像.1 mm19的图像. ОКТ использует короткую длину когерентности широкополосного источника света для сбора изображений на глубине >1 inqaku lencwadi ebhaliweyo. I-OCT isebenzisa ubude obufutshane bokuhambelana komthombo wokukhanya kwe-broadband ukufumana imifanekiso> 1 mm19 kwindiza ethe nkqo.Nangona kunjalo, le ndlela yokuthelekisa iphantsi ixhomekeke kwingqokelela yokukhanya esasaziweyo ngasemva kunye nokulungiswa komfanekiso kuthintelwe zii-artifacts ze-spekle.I-Photoacoustic endoscopy ivelisa kwimifanekiso ye-vivo esekwe kukwandiswa kwe-thermoelastic ngokukhawuleza kwiithishu emva kokufunxwa kwe-laser pulse eyenza amaza omsindo20. Le ndlela ibonise ubunzulu bomfanekiso> 1 cm kwikholoni yomntu kwi-vivo ukujonga unyango. Le ndlela ibonise ubunzulu bomfanekiso> 1 cm kwikholoni yomntu kwi-vivo ukujonga unyango. Этот подход продемонстрировал глубину визуализации > 1 см в толстой кишке человека in vivo для мониторинга терапии. Le ndlela ibonise ubunzulu bomfanekiso> 1 cm kwikholoni yomntu kwi-vivo yokubeka iliso kunyango.這种方法已经证明在体内人结肠中成像深度> 1 厘米以监测治疗.這种方法已经证明在体内人结肠中成像深度> 1 Этот подход был продемонстрирован на глубине изображения > 1 см в толстой кишке человека in vivo для мониторинга терапии. Le ndlela ibonakaliswe kubunzulu bomfanekiso> 1 cm kwikholoni yomntu kwi-vivo ukujonga unyango.Umahluko uveliswa ikakhulu yi-hemoglobin kwi-vasculature.I-Multiphoton endoscopy yenza imizobo ye-fluorescence ephezulu echaseneyo xa iifotoni ze-NIR ezimbini okanye ngaphezulu zibetha ii-biomolecules zethishu ngaxeshanye21. Le ndlela inokufezekisa ubunzulu bomfanekiso> 1 mm kunye ne-phototoxicity ephantsi. Le ndlela inokufezekisa ubunzulu bomfanekiso> 1 mm kunye ne-phototoxicity ephantsi. Этот подход может обеспечить глубину изображения > 1 мм с низкой фототоксичностью. Le ndlela inokubonelela ngobunzulu bomfanekiso> 1 mm kunye ne-phototoxicity ephantsi.這种方法可以实现>1 毫米的成像深度，光毒性低.這种方法可以实现>1 毫米的成像深度，光毒性低. Этот подход может обеспечить глубину изображения > 1 мм с низкой фототоксичностью. Le ndlela inokubonelela ngobunzulu bomfanekiso> 1 mm kunye ne-phototoxicity ephantsi.Uxinzelelo oluphezulu lwe-laser femtosecond laser pulses luyafuneka kwaye le ndlela ayizange iqinisekiswe ngokweklinikhi ngexesha le-endoscopy.
Kule prototype, iskena senza kuphela ukuphambuka kwecala, ngoko ke inxalenye ye-optical ikwi-horizontal (XY) indiza.Isixhobo siyakwazi ukusebenza kwinqanaba eliphezulu lesakhelo (20 Hz) kunezibuko zegalvanic (12 Hz) kwinkqubo yeCellvizio.Yandisa ireyithi yesakhelo ukunciphisa ii-artifacts ezinyakazayo kunye nokunciphisa ireyithi yesakhelo ukunyusa umqondiso.Isantya esiphezulu kunye ne-algorithms ezenzekelayo iyafuneka ukuze kuncitshiswe izinto ezinkulu ezishukumayo ezibangelwa yintshukumo ye-endoscopic, intshukumo yokuphefumla, kunye nokuhamba kwamathumbu.I-Parametric resonant scanners iboniswe ukuba ifezekise ukufuduswa kwe-axial ngaphezulu kwamakhulu e-microns22. Imifanekiso ingaqokelelwa kwindiza ethe nkqo (XZ), i-perpendicular kumphezulu we-mucosal, ukubonelela ngembono efanayo naleyo ye-histology (H & E). Imifanekiso ingaqokelelwa kwindiza ethe nkqo (XZ), i-perpendicular kumphezulu we-mucosal, ukubonelela ngembono efanayo naleyo ye-histology (H & E). Изображения могут быть получены в вертикальной плоскости (XZ), перпендикулярной поверхности слизистой облачки, чтобы обеников гистологии (H&E). Imifanekiso inokuthatyathwa kwinqwelo-moya ethe nkqo (XZ) i-perpendicular kumphezulu we-mucosal ukubonelela ngomfanekiso ofanayo nakwi-histology (H & E).可以在垂直于粘膜表面的垂直平面(XZ) 中收集图像，以提供与组织学(H&E) 相同的视图.可以在垂直于粘膜表面的垂直平面(XZ) 中收集图像，以提供与组织学(H&E) Изображения могут быть получены в вертикальной плоскости (XZ), перпендикулярной поверхности слизистой облачки, чтобы обеников гистологическом исследовании (H&E). Imifanekiso inokuthatyathwa kwinqwelo-moya ethe nkqo (XZ) i-perpendicular kumphezulu we-mucosal ukubonelela ngomfanekiso ofanayo novavanyo lwe-histological (H & E).Iskena sinokubekwa kwindawo yangemva kwenjongo apho umqadi wokukhanyisa iwela ecaleni kwe-axis ephambili ye-optical ukunciphisa ubuntununtunu kwi-aberration8.Phantse i-diffraction-limited focal volumes inokuphambuka kumhlaba wokujonga ngokungqongqo.Ukuskena kofikelelo ngokungacwangciswanga kunokwenziwa ukujikisa izibonisi kwindawo ezichazwe ngumsebenzisi9.Intsimi yokujonga ingancitshiswa ukugqamisa iindawo ezingafanelekanga zomfanekiso, ukuphucula umlinganiselo wesignali-kwingxolo, umahluko, kunye nomlinganiselo wesakhelo.Iiskena zinokuveliswa ngobuninzi kusetyenziswa iinkqubo ezilula.Amakhulu ezixhobo angenziwa kwi-silicon yafer wafer ukunyusa imveliso yexabiso eliphantsi lokuvelisa ubunzima kunye nokusabalalisa ngokubanzi.
Indlela yokukhanya egotyiweyo inciphisa ubungakanani bencam eqinileyo ye-distal, iyenza kube lula ukusebenzisa i-endoscope njengesixhobo ngexesha le-colonoscopy yesiqhelo.Kwimifanekiso ye-fluorescent ebonisiweyo, iimpawu ze-subcellular ze-mucosa zingabonwa ukwahlula i-adenomas ye-tubular (i-precancerous) kwi-polyps ye-hyperplastic (benign).Ezi ziphumo zibonisa ukuba i-endoscopy inokunciphisa inani le-biopsies engadingekile23.Iingxaki eziqhelekileyo ezinxulumene notyando zinokuncitshiswa, amaxesha okubeka iliso anokuphuculwa, kwaye uhlalutyo lwe-histological lwezilonda ezincinci zinokuncitshiswa.Sikwabonisa kwimifanekiso ye-vivo yezigulana ezinesifo sokukrala kwamathumbu, kuquka i-ulcerative colitis (UC) kunye ne-Crohn's colitis.I-colonoscopy yokukhanya okumhlophe eqhelekileyo ibonelela ngombono we-macroscopic womphezulu we-mucosal kunye namandla alinganiselweyo okuvavanya ngokuchanekileyo ukuphiliswa kwe-mucosal.I-Endoscopy ingasetyenziselwa kwi-vivo ukuvavanya ukusebenza kwezonyango zebhayoloji ezifana ne-anti-TNF24 antibodies.Uvavanyo oluchanekileyo kwi-vivo lunokunciphisa okanye luthintele ukuphindaphinda kwesifo kunye neengxaki ezifana nokuhlinzwa kunye nokuphucula umgangatho wobomi.Akukho ziziphumo ezibi kakhulu eziye zaxelwa kwizifundo zeklinikhi ezinxulumene nokusetyenziswa kwe-endoscopes ene-fluorescein kwi-vivo25. Amandla e-laser kwi-mucosal surface anqunyelwe kwi-<2 mW ukunciphisa umngcipheko wokulimala kwe-thermal kunye nokuhlangabezana neemfuno ze-FDA zengozi engabalulekanga26 nge-21 CFR 812. Amandla e-laser kwi-mucosal surface anqunyelwe kwi-<2 mW ukunciphisa umngcipheko wokulimala kwe-thermal kunye nokuhlangabezana neemfuno ze-FDA zengozi engabalulekanga26 nge-21 CFR 812. Мощность лазера на поверхности слизистой оболочки была ограничена до <2 мВт, чтобы свести к минимуму риск термического поврежденият FDA icandelo lemveliso Ngokukodwa umfanekiso weskrini 21 CFR 812. Amandla e-laser kwi-mucosal surface ayelinganiselwe kwi-<2 mW ukunciphisa umngcipheko wokulimala kwe-thermal kunye nokuhlangabezana neemfuno ze-FDA kumngcipheko ongenakuphikiswa26 phantsi kwe-21 CFR 812.粘膜表面的激光功率限制在<2 mW，以最大限度地降低热损伤风险，并满足FDA 21 CFR 812 对26粘膜表面的激光功率限制在<2 mW Мощность лазера на поверхности слизистой оболочки была ограничена до <2 мВт, чтобы свести к минимуму риск термического поврежденият1бения 1 FDA iprosesa yedatha 2 относительно незначительного риска26. Amandla e-laser kwi-mucosal surface anqunyelwe kwi-<2 mW ukunciphisa umngcipheko wokulimala kwe-thermal kunye nokuhlangabezana neemfuno ze-FDA 21 CFR 812 zengozi engabalulekanga26.
Uyilo lwesixhobo lunokuguqulwa ukuphucula umgangatho womfanekiso.Ii-optics ezikhethekileyo ziyafumaneka ukunciphisa i-spherical aberration, ukuphucula ukusonjululwa komfanekiso kunye nokunyusa umgama wokusebenza.I-SIL inokulungelelaniswa ukuba ihambelane kangcono nesalathisi se-refractive ye-tissue (~ 1.4) ukuphucula ukudibanisa ukukhanya.I-frequency drive inokulungiswa ukwandisa i-angle esecaleni yesikena kunye nokwandisa intsimi yomfanekiso.Ungasebenzisa iindlela ezizenzekelayo ukususa iifreyimu zomfanekiso kunye nentshukumo ebalulekileyo yokunciphisa esi siphumo.I-field-programmable gate array (FPGA) ene-high-speed data acquisition iya kusetyenziswa ukubonelela ngokulungiswa kwe-real-time real-time correction.Uncedo olukhulu lweklinikhi, iindlela ezizenzekelayo kufuneka zilungise ukutshintshwa kwesigaba kunye nezixhobo ezishukumayo zokutolika umfanekiso wexesha lokwenyani.I-monolithic 3-axis parametric resonant scanner inokuphunyezwa ukwazisa i-axial scanning 22. Ezi zixhobo ziphuhliselwe ukuphumeza ukufuduka okuthe nkqo okungazange kubonwe ngaphambili >400 µm ngokulungelelanisa i-frequency drive kulawulo olubonisa ukuthamba/ukuqinisa amandla27. Ezi zixhobo ziphuhliselwe ukuphumeza ukufuduka okuthe nkqo okungazange kubonwe ngaphambili >400 µm ngokulungelelanisa i-frequency drive kulawulo olubonisa ukuthamba/ukuqinisa amandla27. Эти устройства были разработаны для достижения беспрецедентного вертикального смещения > 400 iiveki ezingaphezulu уется смешанной динамикой смягчения/жесткости27. Ezi zixhobo ziyilelwe ukuphumeza ukufuduka okuthe nkqo okungazange kubonwe kwe> 400 µm ngokuseta i-frequency drive kwimowudi ephawulwa ngomxube othambileyo/oqinileyo27.7移27.這些 許备 的开发是為了在 具有 混合 软化 硬化 学 状态下 调整 驱动 0垂直 位移 27. Эти устройства были разработаны для достижения беспрецедентных вертикальных смещений >400 мкм путем настройки частототыва смещений й размягчения/затвердевания27. Ezi zixhobo ziyilelwe ukuphumeza ukufuduka okungazange kubonwe ngaphambili> 400 µm ngokulungelelanisa i-trigger frequency kwimodi yokuthambisa / ukuqina kwekinetics27.Kwixesha elizayo, umfanekiso othe nkqo onqamlezileyo unokunceda ekuboniseni umhlaza wangoko (T1a).I-capacitive sensing circuit inokuphunyezwa ukulandelela ukuhamba kwe-scanner kunye nokulungisa i-shift shift 28.Ukulungelelaniswa kwesigaba esizenzekelayo usebenzisa i-sensor circuit inokutshintsha i-manual instrument calibration ngaphambi kokusetyenziswa.Ukuthembeka kwesixhobo kunokuphuculwa ngokusebenzisa iindlela ezithembekileyo zokutywinwa kwesixhobo ukwandisa inani lemijikelezo yokucubungula.I-teknoloji ye-MEMS ithembisa ukukhawulezisa ukusetyenziswa kwe-endoscopes ukujonga i-epithelium yezitho ezingenanto, ukuxilonga izifo, kunye nokubeka iliso kunyango ngendlela encinci.Ngophuhliso olongezelelweyo, le ndlela intsha yokucinga inokuba sisisombululo esinexabiso eliphantsi ukuze sisetyenziswe njengesincedisi kwii-endoscopes zonyango kuvavanyo olukhawulezileyo lwe-histological kwaye lunokuthi ekugqibeleni luthathe indawo yohlalutyo lwesiqhelo lokugula.
I-ray tracing simulations yenziwa kusetyenziswa i-ZEMAX optical design software (inguqulo ye-2013) ukumisela iiparameters ze-optical optics ezigxininisiweyo.Iikhrayitheriya zoyilo zibandakanya ukusombululwa kwe-axial ekufutshane ne-diffractive, umgama wokusebenza = 0 µm, kunye nebala lokujonga (FOV) ngaphezulu kwe-250 × 250 µm2.Ukuvuselela kwi-wavelength λex = 488 nm, i-fiber yemodi enye (SMF) isetyenzisiwe.I-achromatic doublets isetyenziselwa ukunciphisa ukuhluka kweqoqo le-fluorescence (Umfanekiso 5a).I-beam idlula kwi-SMF kunye ne-mode field diameter ye-3.5 μm kwaye ngaphandle kwe-truncation idlula embindini we-reflector kunye nobubanzi be-aperture ye-50 μm.Sebenzisa i-lens enzima yokuntywila (i-hemispherical) enesalathisi esiphezulu se-refractive (n = 2.03) ukunciphisa isiganeko se-beam spherical aberration kunye nokuqinisekisa uqhagamshelwano olupheleleyo kunye nomphezulu we-mucosal.I-optics egxininisekile inikezela i-NA iyonke = 0.41, apho i-NA = i-nsinα, n isalathiso se-refractive ye-tissue, i-α yi-angle ephezulu yokuguqulwa kwe-beam.I-diffraction-limited lateral and axial resolutions yi-0.44 kunye ne-6.65 µm, ngokulandelelana, usebenzisa i-NA = 0.41, λ = 488 nm, kunye ne-n = 1.3313.Iilensi ezithengiswayo kuphela ezinobubanzi obungaphandle (OD) ≤ 2 mm zicatshangelwe.Umzila we-optical uhlanganiswe, kwaye i-boam eshiya i-SMF idlula kwi-aperture ephakathi ye-scanner kwaye ibonakaliswe emva kwesibuko esisisigxina (0.29 mm ububanzi).Olu lungelelwaniso lwenza mfutshane ubude besiphelo somda oqinileyo ukuququzelela ukudlula phambili kwe-endoscope kumgangatho (3.2 mm ububanzi) umjelo osebenzayo wee-endoscopes zonyango.Olu phawu lwenza kube lula ukuyisebenzisa njengesixhobo ngexesha le-endoscopy yesiqhelo.
Isikhokelo sokukhanya esongwayo kunye nokupakishwa kwe-endoscope.(a) Umqadi wovuselelo uyaphuma kwi-OBC kwaye udlule kwindawo esembindini yeskena.Umqadi uyandiswa kwaye ubonakaliswe kwisibuko sesetyhula esisisigxina sibuyele kwiskena sokuphambuka kwecala.I-optics egxininisekileyo iquka iperi ye-achromatic doublet lens kunye ne-immersion eqinile (i-hemispherical) lens enika uqhagamshelwano kunye nomgangatho we-mucosal.I-ZEMAX 2013 (https://www.zemax.com/) yoyilo lwe-optical kunye ne-ray tracing simulation.(b) Ibonisa indawo yezixhobo ezahlukeneyo zesixhobo, kubandakanywa ifayibha yemowudi enye (SMF), iskena, izibuko, kunye neelensi.I-Solidworks 2016 (https://www.solidworks.com/) yayisetyenziselwa imodeli ye-3D yokupakishwa kwe-endoscope.
I-SMF (#460HP, Thorlabs) enemowudi yentsimi ye-3.5 µm kumgama ongamaza angama-488 nm yasetyenziswa “njengomngxuma” wokuhluza indawo yokukhanya okungagxininiswanga (Fig. 5b).Ii-SMF zifakwe kwiityhubhu zepolymer eziguquguqukayo (#Pebax 72D, Nordson MEDICAL).Ubude obumalunga neemitha ezi-4 busetyenziselwa ukuqinisekisa umgama owaneleyo phakathi kwesigulane kunye nenkqubo yokucinga.Iperi ye-2 mm MgF2 eqatywe iilensi eziphindwe kabini ze-achromatic (#65568, #65567, Edmund Optics) kunye ne-2 mm yelensi ye-hemispherical engafakwanga (#90858, Edmund Optics) zasetyenziselwa ukugxila kwi-beam kunye nokuqokelela fluorescence.Faka ityhubhu yokuphela kwentsimbi (i-4 mm ubude, i-2.0 mm OD, i-ID ye-1.6 mm) phakathi kwe-resin kunye ne-tube yangaphandle ukuhlukanisa i-vibration ye-scanner.Sebenzisa i-adhesives yonyango ukukhusela isixhobo kumanzi omzimba kunye neenkqubo zokuphatha.Sebenzisa iityhubhu yokunciphisa ubushushu ukukhusela izihlanganisi.
Iskena esidibeneyo senziwe kumgaqo we-parametric resonance.Faka i-50 µm indawo yokuvula embindini wesibonisi ukusasaza umqadi wovuselelo.Ukusebenzisa iseti ye-quadrature comb-driven drives, umqadi owandisiweyo uyaphambukiswa ngokunqamlezayo kwicala le-orthogonal (inqwelomoya ye-XY) kwimowudi yaseLissajous.Ibhodi yokufumana idatha (#DAQ PCI-6115, NI) yasetyenziselwa ukuvelisa iimpawu ze-analog ukulawula iskena.Amandla anikezelwe sisikhulisa sombane esiphezulu (#PDm200, PiezoDrive) ngeengcingo ezibhityileyo (#B4421241, MWS Wire Industries).Yenza i-wiring kwi-armature ye-electrode.Iskena sisebenza kwiifrikhwensi ezikufutshane ne-15 kHz (i-axis ekhawulezayo) kunye ne-4 kHz (i-axis ecothayo) ukufikelela kwi-FOV ukuya kuma-250 µm × 250 µm.Ividiyo inokudubula kwireyithi yesakhelo se-10, 16, okanye i-20 Hz.Ezi reyithi zesakhelo zisetyenziselwa ukungqamanisa umyinge wophiko lwepateni yokuskena yeLissajous, exhomekeke kwixabiso le-X kunye no-Y yophindaphindo lwezihlandlo zeskena29.Iinkcukacha zokurhweba phakathi komlinganiselo wesakhelo, ukusonjululwa kwepixel, kunye nokuxinana kwepateni yokuskena kubonisiwe kumsebenzi wethu wangaphambili14.
I-laser yesimo esiqinileyo (#OBIS 488 LS, ehambelanayo) ibonelela λex = 488 nm ukuvuyisa i-fluorescein yokuchasana komfanekiso (Umfanekiso 6a).I-pigtails ye-Optical ixhunyiwe kwiyunithi yokucoca nge-FC / APC izixhumi (ilahleko ye-1.82 dB) (umzobo 6b).Umqadi uphambukiswa sisibuko se-dichroic (#WDM-12P-111-488 / 500: 600, Oz Optics) kwi-SMF ngenye i-FC / APC isixhumi.Ngokuhambelana ne-21 CFR 812, amandla esiganeko kwi-tissue akhawulelwe kubuninzi be-2 mW ukuhlangabezana neemfuno ze-FDA kumngcipheko ongenakuphikiswa.I-Fluorescence yagqithiswa kwisibuko se-dichroic kunye nesihlungi sokudlulisa ixesha elide (#BLP01-488R, Semrock).I-Fluorescence yathunyelwa kwi-photomultiplier tube (PMT) detector (#H7422-40, Hamamatsu) nge-FC/PC isiqhagamshelene usebenzisa i- ~ 1 m ubude befiber yemultimode ene-50 µm core diameter.Iimpawu zeFluorescent zandiswa ngesantya esiphezulu seamplifier yangoku (#59-179, Edmund Optics).I-software ekhethekileyo (i-LabVIEW 2021, NI) iphuhliselwe ukufumana idatha yexesha langempela kunye nokwenziwa komfanekiso.Amandla e-laser kunye nezicwangciso zokuzuza kwe-PMT zichongwa yi-microcontroller (#Arduino UNO, Arduino) usebenzisa ibhodi yesekethe eprintiweyo ekhethekileyo.I-SMF kunye neengcingo ziphela kwiikhonkco kwaye zidibanise kwi-fiber optic (F) kunye neengcingo (W) izibuko kwisikhululo sesiseko (Umfanekiso 6c).Inkqubo yokucinga iqulethwe kwinqwelo ephathekayo (Umfanekiso 6d). I-transformer yodwa yasetyenziselwa ukunciphisa ukuvuza kwangoku ukuya ku-<500 μA. I-transformer yodwa yasetyenziselwa ukunciphisa ukuvuza kwangoku ukuya ku-<500 μA. Для ограничения тока утечки до <500 мкА использовался изолирующий трансформатор. Isiguquli sokwahlula sisetyenziselwe ukunciphisa ukuvuza kwangoku ukuya ku-<500 µA.使用隔离变压器将泄漏电流限制在<500 μA. <500 μA. Используйте изолирующий трансформатор, чтобы ограничить ток утечки до <500 мкА. Sebenzisa isiguquli sokwahlula ukunqanda ukuvuza kwangoku ukuya ku-<500µA.
inkqubo yokubona.(a) I-PMT, i-laser kunye ne-amplifier zikwisikhululo esisezantsi.(b) Kwibhanki yokucoca, i-laser (eluhlaza okwesibhakabhaka) iqhuba intambo ye-fiber optic nge-FC / APC isixhumi.I-beam igxothwa ngesibuko se-dichroic (DM) kwi-fiber mode eyodwa (SMF) nge-second FC / APC isixhumi.I-Fluorescence (eluhlaza) ihamba nge-DM kunye nesihluzo sokudlula ixesha elide (LPF) ukuya kwi-PMT nge-multimode fiber (MMF).(c) Isiphelo esikufuphi se-endoscope siqhagamshelwe kwi-fiber optic (F) kunye neengcingo (W) zezibuko zesikhululo sesiseko.(d) I-Endoscope, imonitha, isitishi esisisiseko, ikhompyutha, kunye ne-indoskophu yokwahlula kwinqwelo ephathwayo.(a, c) I-Solidworks 2016 yayisetyenziselwa imodeli ye-3D ye-imaging system kunye namacandelo e-endoscope.
Usombululo olusecaleni kunye ne-axial ye-optics egxininisekileyo yalinganiswa ukusuka kwindawo yosasazo lwe-microspheres ekhanyayo (#F8803, Thermo Fisher Scientific) 0.1 µm ububanzi.Qokelela imifanekiso ngokuguqulela i-microsphere ngokuthe tye kwaye ngokuthe nkqo kwi-1 µm amanyathelo usebenzisa inqanaba lomgca (# M-562-XYZ, DM-13, Newport).Isitaki somfanekiso usebenzisa i-ImageJ2 ukufumana imifanekiso enqamlezileyo yeemicrospheres.
I-software ekhethekileyo (i-LabVIEW 2021, NI) iphuhliselwe ukufumana idatha yexesha langempela kunye nokwenziwa komfanekiso.Kwikhiwane.7 ibonisa amagqabantshintshi eendlela ezisetyenziswayo ekuqhubeni inkqubo.I-interface yomsebenzisi iqulethe i-data acquisition (DAQ), ipaneli ephambili kunye nephaneli yokulawula.Iphaneli yokuqokelela idatha isebenzisana nephaneli ephambili ukuqokelela nokugcina idatha eluhlaza, ukubonelela ngegalelo kwimimiselo yokuqokelela idatha, kunye nokulawula izicwangciso zomqhubi we-scanner.Iphaneli ephambili ivumela umsebenzisi ukuba akhethe ubumbeko olufunwayo lokusebenzisa i-endoscope, kubandakanya isiginali yokulawula iskena, ireyithi yesakhelo sevidiyo, kunye neeparamitha zokufumana.Le ndawo yolawulo ikwavumela umsebenzisi ukuba abonise kwaye alawule ukuqaqamba kunye nokuchasana komfanekiso.Ukusebenzisa idatha ekrwada njengegalelo, i-algorithm ibala ukusetwa kwenzuzo efanelekileyo ye-PMT kwaye ilungelelanise ngokuzenzekelayo le parameter usebenzisa inkqubo yokulawula impendulo ye-proportional-integral (PI) 16.Ibhodi yokulawula isebenzisana nebhodi ephambili kunye nebhodi yokufumana idatha ukulawula amandla e-laser kunye nokuzuza kwe-PMT.
Inkqubo yoyilo lwesoftware.Ujongano lomsebenzisi luquka iimodyuli (1) ukufunyanwa kwedatha (DAQ), (2) iphaneli ephambili kunye (3) nephaneli yokulawula.Ezi nkqubo ziqhutywa ngaxeshanye kwaye zinxibelelana ngemigca yemiyalezo.Isitshixo yi-MEMS: INkqubo yeMicroelectromechanical, TDMS: Ukuhamba koLawulo lweDatha yoBugcisa, i-PI: i-Proportional Integral, i-PMT: I-Photomultiplier.Iifayile zemifanekiso kunye nevidiyo zigcinwa kwiifomathi ze-BMP kunye ne-AVI, ngokulandelelanayo.
I-algorithm yokulungiswa kwesigaba isetyenziselwa ukubala ukusasazwa kokuqina kwe-pixel yomfanekiso kumaxabiso ahlukeneyo esigaba ukumisela elona xabiso liphezulu elisetyenziselwa ukulola umfanekiso.Ukulungiswa kwexesha langempela, uluhlu lokuskena kwesigaba ± 2.86 ° kunye nesinyathelo esikhulu se-0.286 ° ukunciphisa ixesha lokubala.Ukongeza, ukusebenzisa iinxalenye zomfanekiso kunye neesampuli ezimbalwa kunciphisa ngakumbi ixesha lokubala isakhelo somfanekiso ukusuka kwimizuzwana ye-7.5 (1 Msample) ukuya kwimizuzwana eyi-1.88 (250 Ksample) kwi-10 Hz.Ezi parameters zegalelo zikhethwe ukunika umgangatho owaneleyo womfanekiso kunye ne-latency encinci ngexesha lokucinga kwe-vivo.Imifanekiso ephilayo kunye neevidiyo zirekhodwa kwiifomathi ze-BMP kunye ne-AVI, ngokulandelelanayo.Idatha eluhlaza igcinwe kwiFomathi yokuLawula iDatha yoBugcisa (TMDS).
I-post-processing yemifanekiso ye-vivo yokuphucula umgangatho kunye ne-LabVIEW 2021. Ukuchaneka kulinganiselwe xa usebenzisa i-algorithms yokulungiswa kwesigaba ngexesha le-imaging ye-vivo ngenxa yexesha elide lokubala elifunekayo.Kuphela iindawo zemifanekiso eziqingqiweyo kunye namanani esampula asetyenziswayo.Ukongeza, i-algorithm ayisebenzi kakuhle kwimifanekiso enezixhobo ezishukumayo okanye umahluko ophantsi kwaye ikhokelela kwiimpazamo zokubala zesigaba30.Izakhelo zomntu ngamnye ezinomahluko ophezulu kwaye akukho zixhobo zishukumayo zikhethwe ngesandla kwisigaba solungiso olucokisekileyo kunye noluhlu lwesigaba sokuskena ± 0.75 ° kumanyathelo ayi-0.01 °.Yonke indawo yomfanekiso isetyenzisiwe (umzekelo, iSampulu e-1 yomfanekiso orekhodwe nge-10 Hz).Itheyibhile ye-S2 ichaza iiparameters zemifanekiso ezisetyenziselwa ixesha langempela kunye ne-post-processing.Emva kokulungiswa kwesigaba, isihluzo esiphakathi sisetyenziselwa ukunciphisa ngakumbi ingxolo yomfanekiso.Ukukhanya kunye nokuchasana kuphuculwe ngakumbi ngokwandisa i-histogram kunye nokulungiswa kwe-gamma31.
Ulingo lwezonyango luye lwavunywa yiBhodi yokuHlola amaziko oNyango eMichigan kwaye lwaqhutywa kwiSebe leeNkqubo zezoNyango.Olu pho nonongo lubhaliswe kwi-intanethi nge-ClinicalTrials.gov (NCT03220711, umhla wokubhalisa: 07/18/2017).Iikhrayitheriya zokubandakanywa zibandakanya izigulane (ezineminyaka eyi-18 ukuya kwi-100 iminyaka) kunye ne-colonoscopy ekhethiweyo ecetywayo ngaphambili, ingozi eyongeziweyo yomhlaza we-colorectal, kunye nembali yesifo sesibindi esivuthayo.Imvume enolwazi ifunyenwe kwisifundo ngasinye esivumileyo ukuthatha inxaxheba.Iikhrayitheriya zokukhutshelwa ngaphandle yayizizigulana ezikhulelweyo, ezazine-hypersensitivity eyaziwayo kwi-fluorescein, okanye zaziphantsi kwechemotherapy okanye unyango lwemitha.Olu phononongo lubandakanya izigulane ezilandelelanayo ezicwangciselwe i-colonoscopy yesiqhelo kwaye yayimele iZiko lezoNyango laseMichigan.Uphononongo luqhutywe ngokuhambelana neSibhengezo saseHelsinki.
Phambi kotyando, linganisa i-endoskopu usebenzisa i-10 µm amaso e-fluorescent (#F8836, Thermo Fisher Scientific) efakwe kwimold ye-silicone.I-silicone sealant eguquguqukayo (#RTV108, Momentive) yagalelwa kwi-3D eprintiweyo ye-8 cm3 yokubumba yeplastiki.Lahla amaso e-fluorescent phezu kwe-silicone kwaye ushiye de i-medium medium yome.
Ikholoni yonke ihlolwe kusetyenziswa ikholonoscope yonyango eqhelekileyo (i-Olympus, CF-HQ190L) kunye nokukhanya okumhlophe kokukhanya.Emva kokuba i-endoscopist inqume indawo yesi sifo, indawo ihlanjwa nge-5-10 ml ye-acetic acid eyi-5%, kwaye emva koko ngamanzi angcolileyo ukususa i-mucus kunye ne-debris.I-5 ml ye-dose ye-5 mg / ml ye-fluorescein (i-Alcon, i-Fluorescite) ifakwe nge-intravenously okanye i-sprayed topical on the mucosa isebenzisa i-cannula eqhelekileyo (M00530860, i-Boston Scientific) eyagqithiswa kumjelo osebenzayo.
Sebenzisa isinkcenkceshelo ukugungxula idayi engaphezulu okanye inkunkuma kumphezulu we-mucosal.Susa i-nebulizing catheter kwaye ugqithise i-endoscope kumjelo osebenzayo ukufumana imifanekiso ye-ante-mortem.Sebenzisa isikhokelo se-endoscopic esikwibala elibanzi ukubeka incam ekude kwindawo ekujoliswe kuyo. Ixesha elipheleleyo elisetyenziselwa ukuqokelela imifanekiso ye-confocal yayingu <10 min. Ixesha elipheleleyo elisetyenziselwa ukuqokelela imifanekiso ye-confocal yayingu <10 min. Общее время, затраченное на сбор конфокальных изображений, составило <10 мин. Ixesha elipheleleyo elithathiweyo ukuqokelela imifanekiso ye-confocal yayingu <10 min.Ixesha elipheleleyo lokufumana imifanekiso ye-confocal yayingaphantsi kwemizuzu ye-10.Ividiyo yokukhanya emhlophe ye-Endoscopic yacutshungulwa kusetyenziswa inkqubo ye-imaging ye-Olympus EVIS EXERA III (CLV-190) kwaye irekhodwe usebenzisa i-Elgato HD irekhoda yevidiyo.Sebenzisa iLabVIEW 2021 ukurekhoda nokugcina iividiyo ze-endoscopy.Emva kokuba i-imaging igqityiwe, i-endoscope iyasuswa kwaye izicubu eziza kubonwa zisikwa kusetyenziswa i-biopsy forceps okanye umgibe. Izicubu zicutshungulwe kwi-histology yesiqhelo (H&E), kwaye zavavanywa yingcali ye-GI pathologist (HDA). Izicubu zicutshungulwe kwi-histology yesiqhelo (H&E), kwaye zavavanywa yingcali ye-GI pathologist (HDA). Ткани были обработаны для обычной гистологии (H&E) и оценены экспертом-патологом желудочно-кишечного тракта (HDA). Izicubu zacutshungulwa kwi-histology yesiqhelo (H&E) kwaye zavavanywa yingcali ye-gastrointestinal pathologist (HDA).对组织进行常规组织学(H&E) 处理，并由专家GI 病理学家(HDA) 进评估。对组织进行常规组织学(H&E) 处理，并由专家GI 病理学家(HDA) 进评估。 Ткани были обработаны для обычной гистологии (H&E) и оценены экспертом-патологом желудочно-кишечного тракта (HDA). Izicubu zacutshungulwa kwi-histology yesiqhelo (H&E) kwaye zavavanywa yingcali ye-gastrointestinal pathologist (HDA).Iimpawu ze-spectral ze-fluorescein ziqinisekisiwe ngokusebenzisa i-spectrometer (i-USB2000 +, i-Ocean Optics) njengoko kuboniswe kuMfanekiso S2.
I-Endoscopes ivalwa emva kokusetyenziswa komntu ngamnye (Umfanekiso 8).Iinkqubo zokucoca zenziwa phantsi kolawulo kunye nokuvunywa kweSebe loLawulo lweNtsholongwane kunye ne-Epidemiology yeZiko lezoNyango laseMichigan kunye neYunithi yaseCentral Sterile Processing. Ngaphambi kophononongo, izixhobo zavavanywa kwaye zaqinisekiswa ukuvalwa inzala yi-Advanced Sterilization Products (ASP, Johnson & Johnson), iqumrhu lezorhwebo elibonelela ngothintelo losulelo kunye neenkonzo zokuqinisekisa inzala. Ngaphambi kophononongo, izixhobo zavavanywa kwaye zaqinisekiswa ukuvalwa inzala yi-Advanced Sterilization Products (ASP, Johnson & Johnson), iqumrhu lezorhwebo elibonelela ngothintelo losulelo kunye neenkonzo zokuqinisekisa inzala. Перед исследованием инструменты были протестированы и одобрены для стерилизации компанией Advanced Sterilization Products (ASP, Johnson & Johnson), коммерческой ордцеской, и попрофилактике инфекций и проверке стерилизации. Phambi kokuphononongwa, izixhobo ziye zavavanywa kwaye zamkelwa ukuvalwa inzala ziiMveliso eziPhezulu zokuvala inzala (ASP, Johnson & Johnson), umbutho wezorhwebo obonelela ngothintelo losulelo kunye neenkonzo zokuqinisekisa ukuvala inzala. Перед исследованием инструменты были стерилизованы и проверены IiMveliso zokuSebenza eziNgqongileyo (ASP, uJohnson & Johnson), коммерческой организацией, которая предостакефипля предоставлязей й и проверке стерилизации. Izixhobo ziye zavalwa inzala zaza zahlolwa phambi kokuba zifundwe yi-Advanced Sterilization Products (ASP, Johnson & Johnson), umbutho wezorhwebo obonelela ngothintelo losulelo kunye neenkonzo zokuqinisekisa ukuvala inzala.
Isixhobo sokurisayikilisha.(a) Ii-Endoscopes zifakwa kwiitreyi emva kokwenziwa inzala ngakunye kusetyenziswa inkqubo ye-STERRAD.(b) I-SMF kunye neengcingo ziphelisiwe nge-fiber optic kunye neziqhagamshelo zombane, ngokulandelelanayo, ezivaliweyo ngaphambi kokulungiswa kwakhona.
Coca ii-endoscopes ngokwenza oku kulandelayo: (1) sula i-endoscope ngelaphu elingena-lint elifakwe kwisicoci se-enzymatic ukusuka kwiproximal ukuya kwi-distal;(2) Gxininisa isixhobo kwisisombululo se-enzymatic detergent imizuzu emi-3 ngamanzi.ilaphu elingena-lint.Izixhumi zombane kunye nefiber optic zigqunyiwe kwaye zisuswe kwisisombululo;(3) I-endoscope isongiwe kwaye ifakwe kwi-tray yesixhobo sokwenziwa inzala usebenzisa i-STERRAD 100NX, i-hydrogen peroxide yegesi yeplasma.ubushushu obuphantsi nokobume bokufuma obuphantsi.
Iiseti zedatha ezisetyenzisiweyo kunye/okanye ezihlalutyiweyo kuphononongo lwangoku ziyafumaneka kubabhali abachaphazelekayo ngesicelo esinengqiqo.
I-Pilonis, ND, Januszewicz, W. & di Pietro, M. Confocal laser endomicroscopy kwi-endoscopy ye-gastro-intestinal: Imiba yobugcisa kunye nezicelo zeklinikhi. I-Pilonis, ND, Januszewicz, W. & di Pietro, M. Confocal laser endomicroscopy kwi-endoscopy ye-gastro-intestinal: Imiba yobugcisa kunye nezicelo zeklinikhi.I-Pilonis, ND, Januszewicz, V. i di Pietro, M. Confocal laser endomicroscopy kwi-endoscopy yesisu: imiba yobugcisa kunye nesicelo seklinikhi. I-Pilonis, ND, Januszewicz, W. & di Pietro, M. Pilonis, ND, Januszewicz, W. & di Pietro, M. 共载肠分别在在共公司设计在机机：Imiba yobuchwephesha kunye nezicelo zeklinikhi.I-Pilonis, ND, Januszewicz, V. i di Pietro, M. Confocal laser endoscopy kwi-endoscopy yesisu: imiba yobugcisa kunye nezicelo zeklinikhi.translation heparin emathunjini.7, 7 (2022).
Al-Mansour, MR et al.Uhlalutyo loKhuseleko kunye nokuSebenza kwe-SAGES TAVAC Confocal Laser Endomicroscopy.Ukusebenza.I-Endoscopy 35, 2091-2103 (2021).
Fugazza, A. et al.I-Confocal laser endoscopy kwizifo zesisu kunye ne-pancreatobiliary: ukuphononongwa okucwangcisiweyo kunye nohlalutyo lwe-meta.Inzululwazi yezoNyango.itanki yokugcina.yangaphakathi 2016, 4638683 (2016).