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I-Confocal laser endoscopy iyindlela entsha ye-real-time optical biopsy.Izithombe ze-fluorescent zekhwalithi ye-histological zingatholakala ngokushesha ku-epithelium yezitho ezingenalutho.Okwamanje, ukuskena kwenziwa cishe ngamathuluzi asekelwe ku-probe avame ukusetshenziswa emisebenzini yomtholampilo, nokuguquguquka okulinganiselwe ekulawuleni okugxilile.Sibonisa ukusetshenziswa kwesikena se-parametric resonant esifakwe ekugcineni kwe-endoscope ukuze senze ukuchezuka kwe-lateral kwesivinini esikhulu.Imbobo ifakwe phakathi nendawo yesibonisi ukuze kufihlwe indlela yokukhanya.Lo mklamo wehlisa usayizi wensimbi ube ngu-2.4 mm ububanzi no-10 mm ubude, okuwuvumela ukuthi udluliselwe phambili ngomzila osebenzayo wama-endoscopes wezokwelapha ajwayelekile.I-compact lens ihlinzeka ngokulungiswa kwe-lateral ne-axial okungu-1.1 no-13.6 µm, ngokulandelanayo.Ibanga lokusebenza elingu-0 µm kanye nenkundla yokubuka engu-250 µm × 250 µm kufinyelelwa ngamanani ozimele afika ku-20 Hz.Injabulo ku-488 nm ijabulisa i-fluorescein, udayi ogunyazwe yi-FDA wokugqama kwezicubu eziphezulu.Ama-Endoscopes acutshungulwe kabusha imijikelezo engu-18 ngaphandle kokwehluleka kusetshenziswa izindlela zokuvala inzalo ezigunyazwe ngokomtholampilo.Izithombe ezikhanyayo zitholwe ku-colonic mucosa evamile, i-tubular adenomas, i-hyperplastic polyps, i-ulcerative colitis, kanye ne-Crohn's colitis ngesikhathi se-colonoscopy evamile.Amaseli angawodwa angabonakala, okuhlanganisa ama-colonocyte, amaseli e-goblet, namaseli avuthayo.Izici ze-mucosal ezifana nezakhiwo ze-crypt, i-crypt cavities, ne-lamina propria zingahlukaniswa.Ithuluzi lingasetshenziswa njengesengezo se-endoscopy evamile.
I-Confocal laser endoscopy iyindlela entsha yokucabanga eyenzelwe ukusetshenziswa emtholampilo njengesengezo se-endoscopy1,2,3.Lezi zinsimbi eziguquguqukayo, ezixhunywe nge-fiber-optic zingasetshenziswa ukuthola izifo kumaseli e-epithelial ahambisana nezitho ezingenalutho, njengekholoni.Lolu ngqimba oluncane lwezicubu lusebenza kakhulu futhi luwumthombo wezinqubo eziningi zezifo ezinjengomdlavuza, ukutheleleka, nokuvuvukala.I-Endoscopy ingafinyelela ukulungiswa kwe-subcellular, inikeze isikhathi sangempela, ikhwalithi ye-histological ezithombeni ze-vivo ukusiza odokotela benze izinqumo zomtholampilo.I-biopsy yezicubu zomzimba ithwala ingozi yokopha nokubhobozwa.Izifanekiso ze-biopsy eziningi kakhulu noma ezimbalwa kakhulu zivame ukuqoqwa.Isampula ngayinye ekhishiwe inyusa izindleko zokuhlinzwa.Kuthatha izinsuku ezimbalwa ukuthi isampula lihlolwe udokotela wezifo.Phakathi nezinsuku zokulinda imiphumela ye-pathology, iziguli zivame ukuba nokukhathazeka.Ngokuphambene nalokho, ezinye izindlela zokucabanga ngomtholampilo ezifana ne-MRI, CT, PET, SPECT, kanye ne-ultrasound azikho ukulungiswa kwendawo kanye nesivinini sesikhashana esidingekayo ukuze kubonakale ngeso lengqondo izinqubo ze-epithelial ku-vivo ngesikhathi sangempela, ukulungiswa kwe-subcellular.
Ithuluzi elisekelwe ku-probe (Cellvizio) okwamanje livame ukusetshenziswa emitholampilo ukwenza "i-optical biopsy".Idizayini isuselwe ku-fiber optic bundle4 ehambisana ngendawo eqoqa futhi idlulise izithombe ze-fluorescent.I-fiber core eyodwa isebenza "njengembobo" yokuhlunga ukukhanya okungagxilile ukuze kulungiswe ama-subcellular.Ukuskena kwenziwa cishe kusetshenziswa i-galvanometer enkulu, enkulu.Lokhu kunikezwa kukhawulela ikhono lethuluzi lokulawula ukugxila.Isiteji esifanele se-epithelial carcinoma yakuqala sidinga ukubonwa ngaphansi kwethishu ukuze kuhlolwe ukuhlasela nokunquma ukwelashwa okufanele.I-Fluorescein, i-ejenti yokuqhathanisa egunyazwe i-FDA, isetshenziswa ngomthambo ukuze kugqanyiswe izici zesakhiwo se-epithelium. Lawa ma-endomicroscope anobukhulu obungu-<2.4 mm ububanzi, futhi angadluliselwa phambili kalula ngeshaneli ye-biopsy yama-endoscopes wezokwelapha ajwayelekile. Lawa ma-endomicroscope anobukhulu obungu-<2.4 mm ububanzi, futhi angadluliselwa phambili kalula ngeshaneli ye-biopsy yama-endoscopes wezokwelapha ajwayelekile. Эти эндомикроскопы имеют размеры <2,4 мм в диаметре и могут быть легко проведены через биопсийный канал стандартных медицински медицински. Lawa ma-endomicroscopes anobubanzi obungu-<2.4 mm futhi angadlula kalula kumzila we-biopsy wama-endoscopes wezokwelapha ajwayelekile.Lawa ma-borescopes angaphansi kuka-2.4 mm ububanzi futhi adlula kalula kumzila we-biopsy wama-borescope wezokwelapha ajwayelekile.Lokhu kuvumelana nezimo kuvumela inhlobonhlobo yezicelo zomtholampilo futhi kuzimele kubakhiqizi be-endoscope.Kuye kwenziwa izifundo eziningi zomtholampilo kusetshenziswa le divayisi yokuthwebula izithombe, okuhlanganisa ukutholwa kusenesikhathi komdlavuza womminzo, wesisu, wekholoni, nowomlomo.Amaphrothokholi ezithombe athuthukisiwe futhi ukuphepha kwenqubo sekusunguliwe.
Amasistimu we-Microelectromechanical (MEMS) ubuchwepheshe obunamandla bokuklama nokwenza izindlela zokuskena ezincane ezisetshenziswa ekugcineni kwama-endoscopes.Lesi sikhundla (esihlobene ne-proximal) sivumela ukuguquguquka okukhulu ekulawuleni indawo yokugxila5,6.Ngokungeziwe ekuchezukeni kwezinhlangothi, indlela ekude ingakwazi futhi ukwenza ukuskena kwe-axial, ukuskena kwenhloso yangemuva, nokuskena kokufinyelela okungahleliwe.Lawa makhono anika amandla ukuphenywa kweseli ye-epithelial ebanzi, okuhlanganisa i-imaging ye-cross-sectional eqondile7, inkambu enkulu yokubuka (i-FOV)8 ukuskena okungahlangani, nokusebenza okuthuthukisiwe ezifundeni ezichazwe umsebenzisi9.I-MEMS ixazulula inkinga enkulu yokupakisha injini yokuskena ngesikhala esilinganiselwe esitholakala ekugcineni kwethuluzi.Uma kuqhathaniswa nama-galvanometers amakhulu, i-MEMS ihlinzeka ngokusebenza okuphezulu ngosayizi omncane, isivinini esikhulu, kanye nokusetshenziswa kwamandla okuphansi.Inqubo elula yokukhiqiza inganyuswa ukuze kukhiqizwe ngobuningi ngezindleko eziphansi.Imiklamo eminingi ye-MEMS ibikwe ngaphambilini10,11,12.Abukho ubuchwepheshe obuthuthukisiwe ngokwanele ukuvumela ukusetshenziswa okubanzi komtholampilo kwe-real-time in vivo imaging ngokusebenzisa umgudu osebenzayo we-endoscope yezokwelapha.Lapha, sihlose ukukhombisa ukusetshenziswa kwesikena se-MEMS ekugcineni kwe-endoscope ukuze kutholwe isithombe somuntu nge-vivo phakathi ne-endoscopy yomtholampilo evamile.
Ithuluzi le-fiber optic lakhiwe kusetshenziswa isithwebuli se-MEMS ekugcineni ukuqoqa izithombe ze-vivo fluorescent zesikhathi sangempela ezinezici ezifanayo ze-histological.I-fiber yemodi eyodwa (i-SMF) ivalwe ngeshubhu ye-polymer eguquguqukayo futhi ijabule ku-λex = 488 nm.Lokhu kumisa kufinyeza ubude bethiphu ekude futhi kuvumele ukuthi idluliselwe phambili ngomzila osebenzayo wama-endoscopes wezokwelapha ajwayelekile.Sebenzisa ithiphu ukuze ubeke phakathi i-optic.Lawa ma-lens aklanyelwe ukuzuza ukucaca kwe-axial ecishe ihluke ngembobo yezinombolo (NA) = 0.41 kanye nebanga lokusebenza = 0 µm13.Amashimu aqondile enziwa ukuze aqondanise ngokunembile i-optics 14. Isikena sipakishwa ku-endoscope enethiphu eqinile ye-distal engu-2.4 mm ububanzi no-10 mm ubude (Fig. 1a).Lezi zilinganiso zivumela ukuthi zisetshenziswe emisebenzini yomtholampilo njengesesekeli ngesikhathi se-endoscopy (Fig. 1b).Amandla aphezulu esigameko se-laser esicutshini ayengu-2 mW.
I-Confocal laser endoscopy (CLE) kanye nezikena ze-MEMS.Isithombe esibonisa (a) insimbi epakishiwe enobukhulu obuqinile bethiphu eyi-distal enobubanzi obungu-2.4 mm nobude obungu-10 mm kanye (b) nomzila oqondile wesiteshi esisebenzayo se-endoscope yezokwelapha evamile (Olympus CF-HQ190L).(c) Ukubuka ngaphambili kwesithwebuli okubonisa isibonisi esinembobo emaphakathi engu-50 µm lapho kudlula khona i-excitation beam.Isithwebuli sifakwe ku-gimbal eshayelwa isethi yamadrayivu e-quadrature comb drive.Imvamisa ye-resonant yedivayisi inqunywa usayizi wesiphethu se-torsion.(d) Ukubuka okuseceleni kwesithwebuli okubonisa isithwebuli esigxunyekwe esitendini esinezintambo ezixhunywe kumahange e-electrode ahlinzeka ngezindawo zokuxhuma amasignali okushayela namandla.
Indlela yokuskena iqukethe isibonisi esifakwe ku-gimbal esishayelwa isethi yama-actuator e-quadrature ashayelwa yikamu ukuze achezukise i-beam eceleni (indiza ye-XY) ngephethini ye-Lissajous (Fig. 1c).Imbobo engu-50 µm ububanzi yaqoshwa phakathi nendawo lapho ugongolo oluvusa amadlingozi ludlula khona.Isithwebuli siqhutshwa ngemvamisa ye-resonant yomklamo, engacushwa ngokushintsha ubukhulu be-torsion spring.Amahange e-Electrode aqoshwe emaphethelweni edivayisi ukuze anikeze amaphuzu okuxhumana amandla namasignali okulawula (Fig. 1d).
Isistimu yokuthwebula ifakwe enqoleni ephathekayo engagingqika ingene egumbini lokuhlinza.I-graphical interface yomsebenzisi yakhelwe ukusekela abasebenzisi abanolwazi oluncane lwezobuchwepheshe, njengodokotela nabahlengikazi.Hlola mathupha imvamisa yedrayivu yesithwebuli, imodi ye-beamform, nesithombe se-FOV.
Ubude obubonke be-endoscope bucishe bube ngu-4m ukuvumela ukudlula okuphelele kwamathuluzi kushaneli yokusebenza ye-endoscope yezokwelapha evamile (1.68m), enobude obengeziwe bokulawuleka.Ekupheleni kwe-endoscope, i-SMF nezintambo zinqanyulwa kuzixhumi ezixhumeka kumachweba we-fiber optic kanye nezintambo zesiteshi esiyisisekelo.Ukufakwa kuqukethe i-laser, iyunithi yokuhlunga, isikhulisi samandla kagesi aphezulu kanye nomtshina we-photomultiplier (PMT).I-amplifier inikeza amandla kanye namasignali okushayela kusithwebuli.Iyunithi yesihlungi esibonakalayo ihlanganisa injabulo ye-laser ku-SMF futhi idlulisele i-fluorescence ku-PMT.
Ama-Endoscopes acutshungulwa kabusha ngemva kwenqubo ngayinye yomtholampilo kusetshenziswa inqubo yokuvala inzalo ye-STERRAD futhi angamelana nemijikelezo eyi-18 ngaphandle kokwehluleka.Ngesixazululo se-OPA, azikho izimpawu zomonakalo ezibonwe ngemva kwemijikelezo engaphezu kwe-10 yokubulala amagciwane.Imiphumela ye-OPA isebenze kangcono kune-STERRAD, iphakamisa ukuthi impilo yama-endoscopes inganwetshwa ngokubulala amagciwane okusezingeni eliphezulu kunokuphinda kubulawe inzalo.
Ukulungiswa kwesithombe kwanqunywa ngomsebenzi wokusabalalisa amaphuzu kusetshenziswa ubuhlalu be-fluorescent obunobubanzi obungu-0.1 μm.Ngokulungiswa kwe-lateral ne-axial, ububanzi obugcwele kuhhafu wobukhulu (FWHM) obungu-1.1 no-13.6 µm, ngokulandelanayo, bulinganiswe (Fig. 2a, b).
Izinketho zesithombe.Ukulungiswa kwe-lateral (a) ne-axial (b) ye-optics egxilile ibonakala ngomsebenzi wokusabalalisa iphuzu (PSF) olinganiswa kusetshenziswa ama-microsphere e-fluorescent anobubanzi obungu-0.1 μm.Ububanzi obugcwele obulinganisiwe kuhhafu wobukhulu (FWHM) babungu-1.1 no-13.6 µm, ngokulandelanayo.Okufakiwe: Ukubuka okunwetshiwe kwe-microsphere eyodwa ku-transverse (XY) kanye nezikhombisi-ndlela ze-axial (XZ) ziyaboniswa.(c) Isithombe se-Fluorescent esitholwe kumugqa oqondiwe ojwayelekile (USAF 1951) (oval obomvu) obonisa ukuthi amaqembu 7-6 angaxazululwa ngokucacile.(d) Isithombe sika-10 µm ububanzi obuhlakazekile be-fluorescent microsphere esibonisa inkambu yesithombe yokubuka engu-250 µm×250 µm.Ama-PSF ku-(a, b) akhiwe kusetshenziswa i-MATLAB R2019a (https://www.mathworks.com/).(c, d) Izithombe ze-Fluorescent zaqoqwa kusetshenziswa i-LabVIEW 2021 (https://www.ni.com/).
Izithombe ezikhanyayo ezivela kumalensi okucaca okujwayelekile zihlukanisa ngokucacile isethi yamakholomu emaqenjini angu-7-6, agcina ukucaca okuseceleni okuphezulu (Fig. 2c).Inkambu yokubuka (i-FOV) ka-250 µm × 250 µm yanqunywa kusukela ezithombeni zobuhlalu obuyi-fluorescent obuyi-10 µm ububanzi obuhlakazwe kuma-coverlip (Fig. 2d).
Indlela ezenzakalelayo yokulawula inzuzo ye-PMT kanye nokulungiswa kwesigaba kusetshenziswa kusistimu yokucabanga yomtholampilo ukuze kuncishiswe ama-artifacts anyakazayo asuka kuma-endoscope, i-colon peristalsis, nokuphefumula kwesiguli.Ukwakhiwa kabusha kwesithombe nokucubungula ama-algorithms achazwe ngaphambilini14,15.Inzuzo ye-PMT ilawulwa yisilawuli se-proportional-integral (PI) ukuvimbela ukugcwala kwesikhala16.Isistimu ifunda ubukhulu be-pixel intensity yozimele ngamunye, ibala izimpendulo ezilinganayo neziphelele, futhi inquma amanani okuzuza kwe-PMT ukuze kuqinisekiswe ukuthi ukushuba kwephikseli kungaphakathi kwebanga elivumelekile.
Phakathi nesithombe se-vivo, ukungafani kwesigaba phakathi kokunyakaza kwesithwebuli nesiginali yokulawula kungabangela ukufiphala kwesithombe.Imiphumela enjalo ingase yenzeke ngenxa yezinguquko kuzinga lokushisa kwedivayisi ngaphakathi komzimba womuntu.Izithombe ezikhanyayo ezimhlophe zibonise ukuthi i-endoscope yayixhumene ne-colonic mucosa evamile ku-vivo (Umfanekiso 3a).Ukufiphala kwamaphikseli angaqondile kahle kungabonwa ezithombeni ezingahluziwe ze-colonic mucosa evamile (Umfanekiso 3b).Ngemuva kokwelashwa ngesigaba esifanele kanye nokulungiswa kokuphambene, izici ezingaphansi kwe-mucosa zingahlukaniswa (Fig. 3c).Ukuze uthole ulwazi olwengeziwe, izithombe ze-confocal ezingahluziwe nezithombe zesikhathi sangempela ezicutshunguliwe ziboniswa ku-Fig. S1, futhi amapharamitha wokwakha kabusha isithombe asetshenziselwa isikhathi sangempela nokucubungula ngemva kwethulwa kuThebula S1 nakuThebula S2.
Ukucutshungulwa kwesithombe.(a) Isithombe se-endoscopic esine-engeli ebanzi esibonisa i-endoscope (E) exhunywe ku-colonic mucosa evamile (N) ukuze kuqoqwe izithombe ze-vivo fluorescent ngemva kokuphathwa kwe-fluorescein.(b) Ukuzulazula kuma-ax angu-X kanye no-Y ngesikhathi sokuskena kungabangela amaphikseli angaqondani kahle ukuthi afiphale.Ngezinjongo zokubonisa, ukushintsha kwesigaba esikhulu kusetshenziswa esithombeni sokuqala.(c) Ngemva kokulungiswa kwesigaba sangemva kokucubungula, imininingwane ye-mucosal ingahlolwa, okuhlanganisa nezakhiwo zokubethela (imicibisholo), nge-lumen emaphakathi (l) ezungezwe i-lamina propria (lp).Amaseli awodwa angahlukaniswa, okuhlanganisa amakholonocyte (c), amaseli egobolondo (g), namaseli avuvukalayo (imicibisholo).Bona ividiyo eyengeziwe 1. (b, c) Izithombe ezicutshungulwe kusetshenziswa i-LabVIEW 2021.
Izithombe ze-confocal fluorescence zitholwe ku-vivo ezifweni ezimbalwa zekholoni ukuze kuboniswe ukusebenza okubanzi komtholampilo kwethuluzi.Ukuthwebula kwe-engeli ebanzi kuqala kwenziwa kusetshenziswa ukukhanya okumhlophe ukuze kutholwe ulwelwesi lwamafinyila olungavamile.I-endoscope ibe isithuthukiswa ngomgudu osebenzayo wekholonoscope futhi ixhunywe ne-mucosa.
I-Wide-field endoscopy, i-confocal endomicroscopy, ne-histology (H&E) izithombe ziboniswa nge-colonic neoplasia, okuhlanganisa i-tubular adenoma kanye ne-hyperplastic polyp. I-Wide-field endoscopy, i-confocal endomicroscopy, ne-histology (H&E) izithombe ziboniswa nge-colonic neoplasia, okuhlanganisa i-tubular adenoma kanye ne-hyperplastic polyp. Широкопольная эндоскопия, конфокальная эндомикроскопия kanye ne-гистологические (H&E) изображения показаны для неоплазии токирублязия токирублязия токирублязия у futhi гиперпластический полип. I-endoscopy yekholoni, i-confocal endomicroscopy, ne-histological (H&E) imaging iboniswa nge-colonic neoplasia, okuhlanganisa i-tubular adenoma kanye ne-hyperplastic polyp.显示结肠肿瘤（包括管状腺瘤和增生性息肉) (H&E) 图像.共 设计 脚肠化 (图像图像 躰化 和 增生性息肉) 的 广角广角 共共 共共共 Широкопольная эндоскопия, конфокальная микроэндоскопия kanye ne-гистологические (H&E) изображения, показывающие опухоскопия опухоскопия опухоскопия опухоскопия мы kanye ne-гиперпластические полипы. I-Broad-field endoscopy, i-confocal microendoscopy, nezithombe ze-histological (H&E) ezibonisa izimila zekholoni, okuhlanganisa i-tubular adenomas kanye nama-polyps aphezulu.I-adenomas ye-tubular ibonise ukulahlekelwa kwezakhiwo ezivamile ze-crypt, ukunciphisa usayizi wamaseli e-goblet, ukuhlanekezela kwe-crypt lumen, nokuqina kwe-lamina propria (Fig. 4a-c).Ama-polyps e-hyperplastic abonise ukwakheka kwe-stellate kwama-crypts, amaseli ambalwa e-goblet, i-lumen efana ne-slit yama-crypts, kanye ne-lamellar crypts engavamile (Fig. 4d-f).
Isithombe sesikhumba esiwugqinsi se-mucosal ku-vivo. I-endoscopy yokukhanya okumhlophe emele, i-confocal endomicroscope, ne-histology (H&E) izithombe ziboniswa (ac) i-adenoma, (df) i-hyperplastic polyp, (gi) i-ulcerative colitis, kanye (jl) ne-Crohn's colitis. I-endoscopy yokukhanya okumhlophe emele, i-confocal endomicroscope, ne-histology (H&E) izithombe ziboniswa (ac) i-adenoma, (df) i-hyperplastic polyp, (gi) i-ulcerative colitis, kanye (jl) ne-Crohn's colitis. Типичные изображения эндоскопии в белом свете, конфокального эндомикроскопа и гистологии (H&E) показаны для (ac) аденомы, конфокального эндомикроскопа и гистологии для (ac) аденомы, (df нного колита и (jl) колита Крона. I-endoscopy yokukhanya okumhlophe okujwayelekile, i-confocal endomicroscope, ne-histology (H&E) izithombe ziboniswa (ac) i-adenoma, (df) i-hyperplastic polyp, (gi) i-ulcerative colitis, kanye (jl) ne-Crohn's colitis.显示了(ac) 腺瘤、(df) 增生性息肉、(gi) 溃疡性结肠炎和(jl) 克罗恩结肠炎的代腨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癬光克罗恩红肠炎的体育性癬光光内肠肠炎性和电视学( H&E ) isithombe. Представлены репрезентативные эндоскопия в белом свете, конфокальная эндоскопия (ac) аденомы, (df) гипезаркоплика l) колита Крона (H&E). I-endoscopy yokukhanya okumhlophe emele, i-confocal endoscopy, kanye ne-histology ye-(ac) adenoma, (df) i-hyperplastic polyposis, (gi) i-ulcerative colitis, kanye (jl) ne-Crohn's colitis (H&E) iyaboniswa.(B) ikhombisa isithombe esihlangene esitholwe ku-vivo kusuka ku-tubular adenoma (TA) kusetshenziswa i-endoscope (E).Lesi silonda esiyingozi sibonisa ukulahlekelwa kwesakhiwo esivamile se-crypt (umcibisholo), ukuhlanekezela kwe-crypt lumen (l), kanye nokuminyana kwe-crypt lamina propria (lp).Ama-Colonocyte (c), ama-goblet cells (g), namaseli avuvukalayo (imicibisholo) nawo angabonakala.Smt.Ividiyo Eyengeziwe 2. (e) ibonisa isithombe esihlangene esitholwe ku-hyperplastic polyp (HP) in vivo.Lesi sici esibucayi sibonisa ukwakheka kwe-crypt ye-stellate (umcibisholo), ilume efana ne-crypt (l), kanye ne-lamina propria (lp) enomumo ongavamile.Ama-Colonocyte (c), ama-goblet cells amaningana (g) namaseli avuvukalayo (imicibisholo) nawo angabonakala.Smt.Ividiyo Eyengeziwe 3. (h) ibonisa izithombe ezihlangene ezitholwe ku-ulcerative colitis (UC) ku-vivo.Lesi simo sokuvuvukala sibonisa ukwakheka kwe-crypt ehlanekezelwe (umcibisholo) namaseli e-goblet avelele (g).Izimpaphe ze-fluorescein (f) zikhishwa kumaseli e-epithelial, okubonisa ukwanda kwemithambo yegazi.Amaseli amaningi avuthayo (imicibisholo) abonakala ku-lamina propria (lp).Smt.Ividiyo Eyengeziwe 4. (k) ibonisa isithombe esihlangene esitholwe ku-vivo endaweni ye-Crohn's colitis (CC).Lesi simo sokuvuvukala sibonisa ukwakheka kwe-crypt ehlanekezelwe (umcibisholo) namaseli e-goblet avelele (g).Izimpaphe ze-fluorescein (f) zikhishwa kumaseli e-epithelial, okubonisa ukwanda kwemithambo yegazi.Amaseli amaningi avuthayo (imicibisholo) abonakala ku-lamina propria (lp).Smt.Ividiyo Eyengeziwe 5. (b, d, h, l) Izithombe zicutshungulwe kusetshenziswa i-LabVIEW 2021.
Isethi efanayo yezithombe zokuvuvukala kwekholoni iyaboniswa, okuhlanganisa i-ulcerative colitis (UC) (Figure 4g-i) kanye ne-Crohn's colitis (Figure 4j-l).Impendulo yokuvuvukala kucatshangwa ukuthi ibonakala ngezakhiwo ze-crypt ezihlanekezelwe ezinamaseli e-goblet aphumayo.I-fluorescein ikhishelwa ngaphandle kwamangqamuzana e-epithelial, okubonisa ukwanda kwemithambo yegazi.Inombolo enkulu yamangqamuzana okuvuvukala ingabonakala ku-lamina propria.
Sibonise ukusetshenziswa komtholampilo kwe-flexible fiber-coupled confocal laser endoscope esebenzisa isikena se-MEMS esibekwe kude ukuze kutholwe isithombe se-vivo.Ngokuvama okuzwakalayo, izilinganiso zozimele ezingafika ku-20 Hz zingafinyelelwa kusetshenziswa imodi yokuskena ye-Lissajous enobukhulu obukhulu ukuze kuncishiswe ama-artifact anyakazayo.Indlela yokubona iyagoqwa ukuze inikeze ukunwetshwa kwebhilidi kanye nembobo yenombolo eyanele ukuze kufinyelelwe ukulungiswa kwecala okungu-1.1 µm.Izithombe ezikhanyayo zekhwalithi ye-histological zitholwe ngesikhathi se-colonoscopy evamile ye-colonic mucosa evamile, i-tubular adenomas, i-hyperplastic polyps, i-ulcerative colitis, kanye ne-Crohn's colitis.Amaseli angawodwa angabonakala, okuhlanganisa ama-colonocyte, amaseli e-goblet, namaseli avuthayo.Izici ze-mucosal ezifana nezakhiwo ze-crypt, i-crypt cavities, ne-lamina propria zingahlukaniswa.Izingxenyekazi zekhompuyutha ezinembayo zenziwe ngomshini omncane ukuze kuqinisekiswe ukuqondana okunembayo kwezingxenye ezithile zokubona nemishini ngaphakathi kwethuluzi lobude obungu-2.4mm x 10mm ubude.Idizayini yokubona yehlisa ubude bethiphu ye-distal eqinile ngokwanele ukuvumela ukudlula okuqondile ngosayizi ojwayelekile (ububanzi obungu-3.2 mm) isiteshi sokusebenza kuma-endoscopes wezokwelapha.Ngakho-ke, kungakhathaliseki ukuthi umenzi, idivayisi ingasetshenziswa kabanzi odokotela endaweni yokuhlala.Ukuthokozisa kwenziwe ku-λex = 488 nm ukuze kujabulise i-fluorescein, udayi ogunyazwe i-FDA, ukuze kutholwe ukugqama okuphezulu.Ithuluzi lacutshungulwa kabusha ngaphandle kwezinkinga emijikelezweni engu-18 kusetshenziswa izindlela zokuvala inzalo ezamukelwa emtholampilo.
Eminye imiklamo emibili yezinsimbi iqinisekisiwe ngokomtholampilo.I-Cellvizio (i-Mauna Kea Technologies) iyi-probe-based confocal laser endoscope (pCLE) esebenzisa inqwaba yezintambo ze-multimode ezihambisanayo ze-fiber optic ukuze iqoqe futhi idlulise izithombe ze-fluorescence1.Isibuko se-galvo esisesiteshini esiyisisekelo senza ukuskena okuhlangene ekugcineni okuseduze.Izingxenye ze-Optical ziqoqwa endizeni evundlile (XY) ngokujula okungu-0 kuye ku-70 µm.Amakhithi e-Microprobe atholakala ukusuka ku-0.91 (inaliti engu-19 G) kuya ku-5 mm ububanzi.Ukulungiswa kwecala kokungu-1 kuye ku-3.5 µm kufinyelelwe.Izithombe ziqoqwe ngesilinganiso sozimele esingu-9 kuya ku-12 Hz nenkambu yokubuka enohlangothi olulodwa ukusuka ku-240 ukuya ku-600 µm.Ipulatifomu isetshenziswe emtholampilo ezindaweni ezahlukahlukene ezihlanganisa i-bile duct, isinye, ikholoni, umminzo, amaphaphu, namanyikwe.I-Optiscan Pty Ltd ithuthukise i-endoscope-based confocal laser endoscope (eCLE) enenjini yokuskena eyakhelwe eshubhuni lokufaka (i-distal end) ye-endoscope yobungcweti (EC-3870K, Pentax Precision Instruments) 17 .Isigaba se-optical senziwa kusetshenziswa i-fiber yemodi eyodwa, futhi ukuskena ohlangothini kwenziwa kusetshenziswa indlela ye-cantilever ngokusebenzisa imfoloko yokulungisa ezwakalayo.I-activator ye-Shape Memory Alloy (Nitinol) isetshenziselwa ukudala i-axial displacement.Ububanzi obuphelele bemojuli ye-confocal buyi-5 mm.Ukuze kugxilwe, kusetshenziswa ilensi ye-GRIN enenombolo yokuvula i-NA = 0.6.Izithombe ezivundlile zitholwe ngokulungiswa kwe-lateral ne-axial okungu-0.7 no-7 µm, ngokulandelanayo, ngenani lozimele elingu-0.8–1.6 Hz kanye nenkambu yokubuka engu-500 µm × 500 µm.
Sibonisa ukulungiswa kwe-subcellular ku-vivo fluorescence imaging etholakala emzimbeni womuntu ngokusebenzisa i-endoscope yezokwelapha kusetshenziswa isikena se-MEMS esisekupheleni.I-Fluorescence inikeza ukugqama kwesithombe okuphezulu, futhi ama-ligand abophezela ekuqondisweni kwendawo yeseli angalebula nge-fluorophores ukuze anikeze ubunikazi be-molecular ukuze kutholakale isifo esithuthukisiwe18.Amanye amasu okubona e-vivo microendoscopy nawo ayathuthukiswa. I-OCT isebenzisa ubude obufushane bokuhlangana obusuka kumthombo wokukhanya kwe-broadband ukuze iqoqe izithombe endizeni eqondile enobunzulu >1 mm19. I-OCT isebenzisa ubude obufushane bokuhlangana obusuka kumthombo wokukhanya kwe-broadband ukuze iqoqe izithombe endizeni eqondile enobunzulu >1 mm19. ОКТ использует короткую длину когерентности широкополосного источника света для сбора изображений в вертикальной плоскости с 19 >. I-OCT isebenzisa ubude obufushane bokuhambisana komthombo wokukhanya kwe-broadband ukuze ithole izithombe endizeni eqondile ene->1 mm ukujula19. 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 мном19 вькополосного. I-OCT isebenzisa ubude obufushane bokuhambisana komthombo wokukhanya kwe-broadband ukuze ithole izithombe > 1 mm19 endizeni eqondile.Kodwa-ke, le ndlela yomehluko ophansi incike ekuqoqweni kokukhanya okuhlakazekile emuva kanye nokulungiswa kwesithombe kukhawulelwe ama-artifacts amachashaza.I-Photoacoustic endoscopy ikhiqiza ku-vivo izithombe ezisuselwe ekwandeni okusheshayo kwe-thermoelastic ezicutshini ngemva kokumuncwa kwe-laser pulse ekhiqiza amaza omsindo20. Le ndlela ibonise ukujula kwesithombe > 1 cm kukholoni yomuntu ku-vivo ukuqapha ukwelashwa. Le ndlela ibonise ukujula kwesithombe > 1 cm kukholoni yomuntu ku-vivo ukuqapha ukwelashwa. Этот подход продемонстрировал глубину визуализации > 1 см в толстой кишке человека in vivo для мониторинга терапии. Le ndlela ibonise ukujula kwesithombe > 1 cm kukholoni yomuntu ku-vivo ukuze kuqashwe ukwelashwa.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 iye yaboniswa ekujuleni kwezithombe > 1 cm kukholoni yomuntu ku-vivo ukuqapha ukwelashwa.Umehluko ukhiqizwa ikakhulukazi i-hemoglobin ku-vasculature.I-Multiphoton endoscopy ikhiqiza izithombe ze-fluorescence ezihluke kakhulu lapho izithombe ezimbili noma ngaphezulu ze-NIR zishaya ama-biomolecule ezicubu ngasikhathi sinye21. Le ndlela ingafinyelela ukujula kwesithombe> 1 mm nge-phototoxicity ephansi. Le ndlela ingafinyelela ukujula kwesithombe> 1 mm nge-phototoxicity ephansi. Этот подход может обеспечить глубину изображения > 1 мм с низкой фототоксичностью. Le ndlela inganikeza ukujula kwesithombe > 1 mm nge-phototoxicity ephansi.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 inganikeza ukujula kwesithombe > 1 mm nge-phototoxicity ephansi.Kudingeka amaphaphu e-laser e-femtosecond aphezulu futhi le ndlela ayizange ifakazelwe ngokomtholampilo ngesikhathi se-endoscopy.
Kulesi sibonelo, isithwebuli senza kuphela ukuchezuka kwezinhlangothi, ngakho ingxenye ye-optical isendizeni evundlile (XY).Idivayisi iyakwazi ukusebenza ngenani lozimele eliphezulu (20 Hz) kunezibuko eziwu-galvanic (12 Hz) ohlelweni lwe-Cellvizio.Khulisa izinga lozimele ukuze unciphise ama-artifact anyakazayo futhi wehlise izinga lozimele ukuze uthuthukise isignali.Ama-algorithms anesivinini esikhulu nazenzakalelayo ayadingeka ukuze kuncishiswe ama-artifact amakhulu anyakazayo abangelwa ukunyakaza kwe-endoscopic, ukunyakaza kokuphefumula, kanye nokuhamba kwamathumbu.Izikena ze-Parametric resonant zibonisiwe ukuthi zifinyelela ukufuduka kwe-axial ngaphezu kwamakhulu ama-microns22. Izithombe zingaqoqwa ngendiza eqondile (XZ), i-perpendicular ebusweni be-mucosal, ukunikeza umbono ofanayo nalowo we-histology (H&E). Izithombe zingaqoqwa ngendiza eqondile (XZ), i-perpendicular ebusweni be-mucosal, ukunikeza umbono ofanayo nalowo we-histology (H&E). Изображения могут быть получены в вертикальной плоскости (XZ), перпендикулярной поверхности слизистой облочки, чтобы обескости, чтобы обескости, I-гистологии (H&E). Izithombe zingathathwa ngendiza eqondile (XZ) ebheke endaweni ye-mucosal ukuze kunikezwe isithombe esifanayo nese-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 облочки, чтобы обескости, чтобы обескости, гистологическом исследовании (H&E). Izithombe zingathathwa ngendiza eqondile (XZ) eqonde endaweni yolwelwesi lwamafinyila ukuze kuhlinzekwe isithombe esifanayo nesokuhlolwa kwe-histological (H&E).Iskena singabekwa endaweni yangemuva kwenhloso lapho i-beam yokukhanyisa iwela eduze kwe-eksisi enkulu yokubona ukuze kuncishiswe ukuzwela ekuphambukeni8.Ivolumu yokugxila ecishe ibe yi-diffraction-limited ingaphambuka phezu kwezinkambu ezinkulu zokubuka.Ukuskena kokufinyelela okungahleliwe kungenziwa ukuchezukisa izibonisi zibe izindawo ezichazwe umsebenzisi9.Inkambu yokubuka ingancishiswa ukuze kugqanyiswe izindawo ezingafanele zesithombe, kuthuthukiswe isilinganiso sesignali-kuya-nomsindo, ukugqama, nezinga lozimele.Izikena zingakhiqizwa ngobuningi kusetshenziswa izinqubo ezilula.Amakhulu amadivaysi angenziwa ku-silicon wafer ngayinye ukwandisa ukukhiqizwa kokukhiqizwa kwenani eliphansi nokusabalalisa okubanzi.
Umzila wokukhanya ogoqiwe wehlisa usayizi wethiphu ye-distal eqinile, okwenza kube lula ukusebenzisa i-endoscope njengesisekeli ngesikhathi se-colonoscopy evamile.Ezithombeni ze-fluorescent ezibonisiwe, izici ezingaphansi kwe-mucosa zingabonakala ukuze zihlukanise i-adenomas ye-tubular (precancerous) kusuka kuma-polyps e-hyperplastic (benign).Le miphumela iphakamisa ukuthi i-endoscopy inganciphisa inani lama-biopsies angadingekile23.Izinkinga ezijwayelekile ezihlobene nokuhlinzwa zingancishiswa, izikhathi zokuqapha zingathuthukiswa, futhi ukuhlaziywa kwe-histological kwezilonda ezincane kungancishiswa.Siphinde sibonisa izithombe ze-vivo zeziguli ezinesifo samathumbu esivuvukalayo, okuhlanganisa i-ulcerative colitis (UC) kanye ne-Crohn's colitis.I-colonoscopy yokukhanya okumhlophe evamile inikeza umbono omkhulu we-mucosal surface enamandla alinganiselwe wokuhlola ngokunembile ukuphulukiswa kwe-mucosal.I-Endoscopy ingasetshenziswa ku-vivo ukuhlola ukusebenza kahle kwemithi yokwelapha ephilayo njengama-anti-TNF24 amasosha omzimba.Ukuhlola okunembile kwe-vivo nakho kunganciphisa noma kuvimbele ukuphinda kwezifo kanye nezinkinga ezinjengokuhlinzwa futhi kuthuthukise izinga lokuphila.Akukho ukusabela okubi okubi okubikiwe ezifundweni zomtholampilo ezihlobene nokusetshenziswa kwe-endoscopes equkethe i-fluorescein ku-vivo25. Amandla e-laser ebusweni be-mucosal ayelinganiselwe ku-<2 mW ukuze kuncishiswe ingozi yokulimala okushisayo futhi ahlangabezane nezimfuneko ze-FDA zengozi engabalulekile26 nge-21 CFR 812 ngayinye. Amandla e-laser ebusweni be-mucosal ayelinganiselwe ku-<2 mW ukuze kuncishiswe ingozi yokulimala okushisayo futhi ahlangabezane nezimfuneko ze-FDA zengozi engabalulekile26 nge-21 CFR 812 ngayinye. Мощность лазера на поверхности слизистой оболочки была ограничена до <2 мВт, чтобы свести к минимуму риск термического поврежденият FDA и совастоватьения и соборовать но незначительного риска26 согласно 21 CFR 812. Amandla e-laser endaweni ye-mucosal ayekhawulelwe ku-<2 mW ukuze kuncishiswe ingcuphe yokulimala okushisayo futhi ahlangabezane nezimfuneko ze-FDA ngengozi enganakeki26 ngaphansi kokungu-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 凙2靎 .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 поврежденит1буму риск термического поврежденит1бостова1 FDA 2 относительно незначительного риска26. Amandla e-laser ebusweni be-mucosal ayelinganiselwe ku-<2 mW ukuze kuncishiswe ingcuphe yokulimala okushisayo futhi ahlangabezane nezimfuneko ze-FDA 21 CFR 812 ngengozi enganakeki26.
Idizayini yensimbi ingashintshwa ukuze kuthuthukiswe ikhwalithi yesithombe.Ama-optics akhethekile ayatholakala ukuze kuncishiswe ukuphuma okuyindilinga, ukuthuthukisa ukucaca kwesithombe nokwenyusa ibanga lokusebenza.I-SIL ingashunwa ukuze ifane kangcono nenkomba ye-refractive yethishu (~1.4) ukuze kuthuthukiswe ukuhlangana kokukhanya.Imvamisa yedrayivu ingashintshwa ukuze kukhuliswe i-engeli esemaceleni yesithwebuli futhi kwandiswe inkambu yokubuka yesithombe.Ungasebenzisa izindlela ezizenzakalelayo ukususa amafreyimu esithombe ngokunyakaza okubalulekile ukuze unciphise lo mphumela.I-field-programmable gate array (i-FPGA) enokutholwa kwedatha enesivinini esiphezulu izosetshenziswa ukuhlinzeka ngokulungiswa kohlaka olugcwele lwesikhathi sangempela sokusebenza okuphezulu okuphezulu.Ukuze uthole usizo olukhulu lomtholampilo, izindlela ezizenzakalelayo kufanele zilungise ukushintsha kwesigaba kanye nama-artifact anyakazayo ukuze kuchazwe isithombe sesikhathi sangempela.Isikena se-monolithic 3-axis parametric resonant singasetshenziswa ukwethula ukuskena kwe-axial 22. Lawa madivayisi athuthukiswe ukuze afinyelele ukugudluzwa okungakaze kubonwe ngaphambili >400 µm ngokulungisa imvamisa yedrayivu ohlelweni oluhlanganisa ukuthambisa/ukuqinisa amandla27. Lawa madivayisi athuthukiswe ukuze afinyelele ukugudluzwa okungakaze kubonwe ngaphambili >400 µm ngokulungisa imvamisa yedrayivu ohlelweni oluhlanganisa ukuthambisa/ukuqinisa amandla27. Эти устройства были разработаны для достижения беспрецедентного вертикального смещения > 400 мкм путем настройки частоты возбраке возбраженные возбражение уется смешанной динамикой смягчения/жесткости27. Lawa madivayisi aklanyelwe ukufeza ukugudluka okungakaze kubonwe ngaphambili kokuthi >400 µm ngokusetha imvamisa yedrayivu kumodi ebonakala ngokuxubile okuthambile/okuqinile27.這些許备的开发是為了通進在具有混合软化/硬化动力学的状态下调整驱动频率來实现 4 Isiqephu 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. Lawa madivayisi adizayinelwe ukufeza ukugudluzwa okungakaze kubonwe ngaphambili >400 µm ngokulungisa imvamisa ye-trigger kumodi ye-kinetics exubile/yokwenza ukuqina27.Ngokuzayo, ukuthwebula izithombe okuqondile kungasiza ekutholeni umdlavuza wakuqala (T1a).Umjikelezo we-capacitive sensing ungasetshenziswa ukuze ulandelele ukunyakaza kwesithwebuli futhi ulungise ukushintshwa kwesigaba sama-28.Ukulinganisa kwesigaba okuzenzakalelayo kusetshenziswa isekethe yenzwa kungangena esikhundleni sokulinganiswa kwensimbi okwenziwa ngesandla ngaphambi kokusetshenziswa.Ukuthembeka kwamathuluzi kungathuthukiswa ngokusebenzisa amasu okuvala amathuluzi athembekile ukuze kwandiswe inani lemijikelezo yokucubungula.Ubuchwepheshe be-MEMS buthembisa ukusheshisa ukusetshenziswa kwama-endoscope ukuze ubone ngeso lengqondo i-epithelium yezitho ezingenalutho, ukuhlonza izifo, nokuqapha ukwelashwa ngendlela engavamile.Ngokuthuthuka okuqhubekayo, le ndlela entsha yokuthwebula ingase ibe yisixazululo esingabizi kakhulu esizosetshenziswa njengesengezo kuma-endoscopes wezokwelapha ukuze kuhlolwe i-histological ngokushesha futhi ingase ithathele indawo ukuhlaziywa kwendabuko kokugula.
Ukulingiswa kokulandelela u-ray kwenziwa kusetshenziswa isofthiwe ye-ZEMAX optical design (inguqulo ka-2013) ukuze kunqunywe amapharamitha we-optics egxilile.Imibandela yokudizayina ihlanganisa i-axial eseduze ne-diffractive resolution, ibanga lokusebenza = 0 µm, kanye nenkambu yokubuka (FOV) enkulu kuno-250 × 250 µm2.Ukuze kujabulise kubude begagasi λex = 488 nm, i-fiber yemodi eyodwa (SMF) isetshenzisiwe.Ukuphinda kabili kwe-Achromatic kusetshenziselwa ukunciphisa ukuhluka kweqoqo le-fluorescence (Umfanekiso 5a).I-beam idlula ku-SMF enobubanzi benkambu yemodi engu-3.5 μm futhi ngaphandle kwe-truncation idlula phakathi nendawo yesibonisi nge-diameter yokuvula engu-50 μm.Sebenzisa ilensi yokucwiliswa eqinile (i-hemispherical) enenkomba ephezulu ye-refractive (n = 2.03) ukuze unciphise ukuphambuka okuyindilinga kwesigameko futhi uqinisekise ukuthintana okugcwele ne-mucosal surface.I-Focusing Optics inikeza inani le-NA = 0.41, lapho i-NA = i-nsinα, n iyinkomba ye-refractive ye-thishu, α iyi-angle ephezulu yokuhlangana kwe-beam.I-diffraction-limited lateral ne-axial resolutions ingu-0.44 kanye no-6.65 µm, ngokulandelanayo, kusetshenziswa i-NA = 0.41, λ = 488 nm, kanye ne-n = 1.3313.Kucatshangelwe amalensi atholakala ngokudayiswa kuphela anobubanzi obungaphandle (OD) ≤ 2 mm.Umzila we-optical uyagoqwa, futhi ugongolo olushiya i-SMF ludlula endaweni emaphakathi yesithwebuli futhi lukhonjiswe emuva ngesibuko esingaguquki (ububanzi obungu-0.29 mm).Lokhu kulungiselelwa kufinyeza ubude bokuphela kwe-distal eqinile ukuze kusize ukudlula phambili kwe-endoscope ngokusebenzisa umzila osebenzayo ojwayelekile (3.2 mm ububanzi) wama-endoscopes wezokwelapha.Lesi sici senza kube lula ukusisebenzisa njengesisekeli ngesikhathi se-endoscopy evamile.
Umhlahlandlela wokukhanya ogoqiwe kanye nokupakishwa kwe-endoscope.(a) I-excitation beam iphuma ku-OBC futhi idlule endaweni emaphakathi yesikena.I-beam iyanwetshwa futhi ibonakale kusukela esibukweni esiyindilinga esingashintshi sibuyela kusikena sokuchezuka kwezinhlangothi.Ama-optics agxilile ahlanganisa amalensi aphindwe kabili e-achromatic kanye nelensi yokucwiliswa eqinile (i-hemispherical) ehlinzeka ngokuthintana nendawo engaphezulu ye-mucosal.I-ZEMAX 2013 (https://www.zemax.com/) yokuklama okubonakalayo nokulingiswa kokulandela umsebe.(b) Ibonisa indawo yezingxenye zensimbi ezihlukahlukene, okuhlanganisa i-fiber yemodi eyodwa (i-SMF), isikena, izibuko, namalensi.I-Solidworks 2016 (https://www.solidworks.com/) isetshenziselwe ukumodela kwe-3D kwephakheji ye-endoscope.
I-SMF (#460HP, Thorlabs) enobubanzi benkambu yemodi engu-3.5 µm kubude begagasi obungu-488 nm yasetshenziswa “njengembobo” yokuhlunga indawo yokukhanya okungagxilile (Fig. 5b).Ama-SMF avalelwe kumashubhu e-polymer aguquguqukayo (#Pebax 72D, Nordson MEDICAL).Ubude obucishe bube ngamamitha angu-4 busetshenziselwa ukuqinisekisa ibanga elanele phakathi kwesiguli kanye nesistimu yokuthwebula izithombe.Ipheya engu-2 mm MgF2 ehlanganiswe amalensi e-achromatic doublet (#65568, #65567, Edmund Optics) kanye nelensi ye-hemispherical engahlanganisiwe engu-2 mm (#90858, Edmund Optics) asetshenziselwa ukugxilisa uhlaka futhi aqoqe i-fluorescence.Faka ishubhu lokugcina lensimbi engagqwali (4 mm ubude, 2.0 mm OD, 1.6 mm ID) phakathi kwe-resin neshubhu elingaphandle ukuze uhlukanise ukudlidliza kwesikena.Sebenzisa izinto ezinamathelayo zezokwelapha ukuze uvikele ithuluzi kuketshezi lomzimba kanye nezinqubo zokuphatha.Sebenzisa amashubhu okunciphisa ukushisa ukuze uvikele izixhumi.
Isithwebuli esihlangene senziwa ngomgomo we-parametric resonance.Faka imbobo engu-50 µm phakathi nendawo yesibonisi ukuze udlulise i-beam yenjabulo.Kusetshenziswa isethi yamadrayivu ashayelwa ikamu le-quadrature, i-beam enwetshiwe iphambukiswa ngokuphambana nendawo ye-orthogonal (indiza ye-XY) ngemodi ye-Lissajous.Ibhodi lokutholwa kwedatha (#DAQ PCI-6115, NI) lisetshenziswe ukukhiqiza amasiginali we-analog ukuze kulawulwe isikena.Amandla anikezwe isikhulisi sikagesi esiphezulu (#PDm200, PiezoDrive) ngezintambo ezincane (#B4421241, MWS Wire Industries).Yenza izintambo ku-armature ye-electrode.Iskena sisebenza kumafrikhwensi asondele ku-15 kHz (i-axis esheshayo) kanye no-4 kHz (i-eksisi ehamba kancane) ukuze kuzuzwe i-FOV kufika ku-250 µm × 250 µm.Ividiyo ingashuthwa ngesilinganiso sozimele esingu-10, 16, noma esingu-20 Hz.Lezi zilinganiso zozimele zisetshenziselwa ukufanisa izinga lokuphinda lephethini yokuskena ye-Lissajous, encike enanini le-X kanye ne-Y ye-excitation frequencies yesikena29.Imininingwane yokuhwebelana phakathi kwezinga lozimele, ukulungiswa kwephikseli, nokuminyana kwephethini yokuskena kuvezwa emsebenzini wethu wangaphambilini14.
I-laser yesimo esiqinile (#OBIS 488 LS, coherent) inikeza i-λex = 488 nm ukuze ijabulise i-fluorescein ngokugqama kwesithombe (Fig. 6a).Ama-pigtails optical axhunywe kuyunithi yokuhlunga ngezixhumi ze-FC/APC (ukulahlekelwa ngu-1.82 dB) (Fig. 6b).Uhlaka luphambukiswa yisibuko se-dichroic (#WDM-12P-111-488/500:600, Oz Optics) ku-SMF ngesinye isixhumi se-FC/APC.Ngokuvumelana ne-21 CFR 812, amandla esigameko esithishu akhawulelwe kubukhulu obungu-2 mW ukuze kuhlangatshezwane nezidingo ze-FDA ngengozi enganakwa.I-fluorescence idlulwe esibukweni se-dichroic kanye nesihlungi eside sokudlulisa (#BLP01-488R, Semrock).I-Fluorescence idluliselwe kumtshina we-photomultiplier tube (PMT) (#H7422-40, Hamamatsu) ngesixhumi se-FC/PC sisebenzisa i-~1 m ubude befayibha ye-multimode enobubanzi obungu-50 µm core.Amasiginali we-Fluorescent akhuliswe nge-amplifier yamanje yesivinini esikhulu (#59-179, Edmund Optics).Isofthiwe ekhethekile (LabVIEW 2021, NI) yenzelwe ukutholwa kwedatha ngesikhathi sangempela kanye nokucutshungulwa kwesithombe.Amandla e-laser nezilungiselelo zenzuzo ye-PMT zinqunywa isilawuli esincane (#Arduino UNO, Arduino) sisebenzisa ibhodi lesifunda eliphrintiwe elikhethekile.I-SMF nezintambo zinqanyulwa kuzixhumi futhi zixhumeke kumachweba we-fiber optic (F) kanye nezintambo (W) esiteshini esiyisisekelo (Umfanekiso 6c).Uhlelo lokuthwebula luqukethwe enqoleni ephathekayo (Umfanekiso 6d). I-isolation transformer yasetshenziswa ukukhawulela ukuvuza kwamanje ku-<500 μA. I-isolation transformer yasetshenziswa ukukhawulela ukuvuza kwamanje ku-<500 μA. Для ограничения тока утечки до <500 мкА использовался изолирующий трансформатор. I-isolation transformer yasetshenziswa ukukhawulela ukuvuza kwamanje 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 i-isolation transformer ukuze ukhawulele ukuvuza kwamanje ku-<500µA.
uhlelo lokubuka.(a) I-PMT, i-laser kanye ne-amplifier kusesiteshini esiyisisekelo.(b) Ebhange lokuhlunga, i-laser (eluhlaza okwesibhakabhaka) ishayela phezu kwentambo ye-fiber optic ngesixhumi se-FC/APC.I-beam iphambukiswa yisibuko se-dichroic (DM) ibe yifayibha yemodi eyodwa (SMF) ngesixhumi sesibili se-FC/APC.I-Fluorescence (eluhlaza) ihamba nge-DM kanye nesihlungi se-long pass (LPF) iye ku-PMT nge-multimode fiber (MMF).(c) Isiphetho esiseduze se-endoscope sixhunywe ku-fiber optic (F) kanye nezimbobo ezinezintambo (W) zesiteshi esiyisisekelo.(d) I-Endoscope, imonitha, isiteshi sesisekelo, ikhompuyutha, kanye ne-isolation transformer enqoleni ephathekayo.(a, c) I-Solidworks 2016 yasetshenziselwa ukumodela kwe-3D kwesistimu yokuthwebula izithombe nezingxenye ze-endoscope.
Ukulungiswa kwe-lateral ne-axial kwe-optics egxilile kukalwa ngomsebenzi wokusabalalisa iphuzu we-fluorescent microspheres (#F8803, Thermo Fisher Scientific) 0.1 µm ububanzi.Qoqa izithombe ngokuhumusha ama-microsphere ngokuvundlile futhi mpo ngezinyathelo ezingu-1 µm usebenzisa isiteji somugqa (# M-562-XYZ, DM-13, Newport).Isitaki sesithombe sisebenzisa i-ImageJ2 ukuze uthole izithombe ezihlukene zama-microsphere.
Isofthiwe ekhethekile (LabVIEW 2021, NI) yenzelwe ukutholwa kwedatha ngesikhathi sangempela kanye nokucutshungulwa kwesithombe.Emkhiwaneni.7 kukhombisa ufingqo lwezinqubo ezisetshenziswa ukusebenzisa uhlelo.Isixhumi esibonakalayo somsebenzisi siqukethe ukutholwa kwedatha (DAQ), iphaneli eyinhloko kanye nephaneli yesilawuli.Iphaneli yokuqoqwa kwedatha isebenzisana nephaneli enkulu ukuze iqoqe futhi igcine idatha eluhlaza, inikeze okokufaka kwezilungiselelo zokuqoqwa kwedatha yangokwezifiso, nokuphatha izilungiselelo zomshayeli wesithwebuli.Iphaneli eyinhloko ivumela umsebenzisi ukuthi akhethe ukucushwa akufisayo ngokusebenzisa i-endoscope, okuhlanganisa isignali yokulawula isithwebuli, izinga lozimele bevidiyo, namapharamitha wokutholwa.Le phaneli futhi ivumela umsebenzisi ukuthi abonise futhi alawule ukukhanya nokugqama kwesithombe.Isebenzisa idatha eluhlaza njengokufakwayo, i-algorithm ibala isilungiselelo senzuzo esiphezulu se-PMT futhi ilungise ngokuzenzakalelayo le pharamitha isebenzisa isistimu yokulawula impendulo ye-proportional-integral (PI)16.Ibhodi lesilawuli lisebenzisana nebhodi elikhulu kanye nebhodi lokutholwa kwedatha ukuze lilawule amandla e-laser kanye nokuzuza kwe-PMT.
Isakhiwo sesoftware yesistimu.Ukuxhumana komsebenzisi kuqukethe amamojula (1) ukutholwa kwedatha (DAQ), (2) iphaneli eyinhloko kanye (3) nephaneli yesilawuli.Lezi zinhlelo zisebenza kanye kanye futhi zixhumana zodwa ngolayini bemiyalezo.Ukhiye uthi i-MEMS: I-Microelectromechanical System, TDMS: Technical Data Control Flow, PI: Proportional Integral, PMT: Photomultiplier.Amafayela ezithombe nawevidiyo agcinwa ngefomethi ye-BMP ne-AVI, ngokulandelana.
I-algorithm yokulungiswa kwesigaba isetshenziselwa ukubala ukusakazeka kokuqina kwephikseli yesithombe ngamavelu esigaba esihlukile ukuze kunqunywe inani eliphezulu elisetshenziselwa ukulola isithombe.Ukulungiswa kwesikhathi sangempela, ibanga lokuskena kwesigaba lingu-±2.86° ngesinyathelo esikhulu kakhulu esingu-0.286° ukuze kuncishiswe isikhathi sokubala.Ngaphezu kwalokho, ukusebenzisa izingxenye zesithombe ezinamasampuli ambalwa kwehlisa futhi isikhathi sokubala sozimele wesithombe sisuka kumasekhondi angu-7.5 (1 Msample) siye kumasekhondi angu-1.88 (250 Ksample) kokuthi 10 Hz.Lezi zinhlaka zokufaka zikhethiwe ukuze zinikeze ikhwalithi yesithombe eyanele enokubambezeleka okuncane phakathi nesithombe se-vivo.Izithombe namavidiyo abukhoma aqoshwa ngefomethi ye-BMP ne-AVI, ngokulandelana.Idatha eluhlaza igcinwe ku-Technical Data Management Flow Format (TMDS).
Ukucutshungulwa kwangemuva kwezithombe ze-vivo ukuze kuthuthukiswe ikhwalithi nge-LabVIEW 2021. Ukunemba kukhawulelwe uma usebenzisa ama-algorithms okulungisa isigaba phakathi nesithombe se-vivo ngenxa yesikhathi eside sokubala esidingekayo.Kusetshenziswa izindawo zesithombe ezikhawulelwe kuphela nezinombolo zesampula.Ngaphezu kwalokho, i-algorithm ayisebenzi kahle ezithombeni ezinama-artifacts anyakazayo noma umehluko ophansi futhi iholela kumaphutha esibalo sesigaba30.Ozimele abangabodwana abanokugqama okuphezulu futhi awekho ama-artifact anyakazayo akhethwe mathupha ukuze kulungiswe kahle isigaba ngebanga lokuskena lesigaba elingu-±0.75° kuzinyathelo ezingu-0.01°.Yonke indawo yesithombe isetshenzisiwe (isb, isampula engu-1 yesithombe esirekhodwe ngo-10 Hz).Ithebula le-S2 linikeza imininingwane ngemingcele yesithombe esetshenziselwe isikhathi sangempela kanye nangemuva kokucubungula.Ngemva kokulungiswa kwesigaba, isihlungi se-median sisetshenziswa ukuze kuqhutshekwe nokunciphisa umsindo wesithombe.Ukukhanya nokugqama kuthuthukiswa nakakhulu ngokwelula kwe-histogram nokulungiswa kwe-gamma31.
Izivivinyo zomtholampilo zigunyazwe Ibhodi Lokubuyekeza Izikhungo Zezokwelapha zaseMichigan futhi zenziwa eMnyangweni Wezinqubo Zokwelashwa.Lolu cwaningo lubhaliswe ku-inthanethi ne-ClinicalTrials.gov (NCT03220711, usuku lokubhalisa: 07/18/2017).Inqubo yokufakwa yayihlanganisa iziguli (ezineminyaka eyi-18 kuye kweyi-100) ezine-colonoscopy ekhethiwe ehleliwe ngaphambili, ingozi eyengeziwe yomdlavuza we-colorectal, kanye nomlando wesifo samathumbu esivuvukalayo.Imvume enolwazi itholwe esifundweni ngasinye esivume ukubamba iqhaza.Imibandela yokukhishwa kwakuyiziguli ezazikhulelwe, ezazinokuzwela okwaziwayo ku-fluorescein, noma ezazilashwa ngamakhemikhali noma ngemisebe.Lolu cwaningo lwaluhlanganisa iziguli ezilandelanayo ezihlelelwe i-colonoscopy evamile futhi lwalumele abantu baseMichigan Medical Center.Ucwaningo lwenziwe ngokuhambisana neSimemezelo saseHelsinki.
Ngaphambi kokuhlinzwa, linganisa i-endoscope usebenzisa ubuhlalu be-fluorescent obungu-10 µm (#F8836, Thermo Fisher Scientific) obufakwe kusikhunta se-silicone.I-translucent silicone sealant (#RTV108, Momentive) ithululwe esikhunjeni sepulasitiki esingu-8 cm3 esiphrintwe nge-3D.Beka ubuhlalu be-fluorescent bamanzi phezu kwe-silicone bese ushiya kuze kube yilapho amanzi omile.
Ikholoni lonke lahlolwa kusetshenziswa i-colonoscope yezokwelapha ejwayelekile (i-Olympus, CF-HQ190L) enokukhanya okumhlophe.Ngemuva kokuthi i-endoscopist inqume indawo yesifo esisolwayo, indawo igezwa ngo-5-10 ml we-acetic acid engu-5%, bese ngamanzi angenalutho ukuze kukhishwe amafinyila kanye nemfucumfucu.Umthamo ongu-5 ml we-5 mg/ml i-fluorescein (i-Alcon, i-Fluorescite) yajovwa ngomthambo noma yafafazwa phezulu ku-mucosa kusetshenziswa i-cannula evamile (M00530860, Boston Scientific) edluliswe esiteshini sokusebenza.
Sebenzisa i-irrigator ukuze ukhiphe udayi owedlulele noma imfucumfucu ebusweni be-mucosal.Susa i-nebulizing catheter futhi udlulise i-endoscope esiteshini esisebenzayo ukuze uthole izithombe ze-ante-mortem.Sebenzisa isiqondiso se-endoscopic yenkundla ebanzi ukuze umise ithiphu elikude endaweni eqondiwe. Isikhathi esiphelele esisetshenziswe ukuqoqa izithombe ezihlangene sasingu-<10 min. Isikhathi esiphelele esisetshenziswe ukuqoqa izithombe ezihlangene sasingu-<10 min. Общее время, затраченное на сбор конфокальных изображений, составило <10 мин. Isikhathi esiphelele esithathiwe ukuze kuqoqwe izithombe zeconfocal sasingu-<10 min.Isikhathi esiphelele sokutholwa kwezithombe ezihlangene besingaphansi kwemizuzu eyi-10.Ividiyo yokukhanya okumhlophe kwe-Endoscopic yacutshungulwa kusetshenziswa isistimu yokucabanga ye-Olympus EVIS EXERA III (CLV-190) futhi yarekhodwa kusetshenziswa irekhoda yevidiyo ye-Elgato HD.Sebenzisa i-LabVIEW 2021 ukuze urekhode futhi ulondoloze amavidiyo e-endoscopy.Ngemuva kokuthi ukuthwebula kuqediwe, i-endoscope iyakhishwa futhi izicubu ezizobonwa zisikwe kusetshenziswa i-biopsy forceps noma ugibe. Izicubu zacutshungulwa i-routine histology (H&E), futhi zahlolwa uchwepheshe we-GI pathologist (HDA). Izicubu zacutshungulwa i-routine histology (H&E), futhi zahlolwa uchwepheshe we-GI pathologist (HDA). Ткани были обработаны для обычной гистологии (H&E) and оценены экспертом-патологом желудочно-кишечного тракта (HDA). Izicubu zacutshungulwa i-routine histology (H&E) futhi zahlolwa uchwepheshe wezifo zesisu (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) and оценены экспертом-патологом желудочно-кишечного тракта (HDA). Izicubu zacutshungulwa i-routine histology (H&E) futhi zahlolwa uchwepheshe wezifo zesisu (HDA).Izakhiwo ze-spectral ze-fluorescein zaqinisekiswa kusetshenziswa i-spectrometer (i-USB2000+, i-Ocean Optics) njengoba kuboniswe kuMfanekiso S2.
Ama-Endoscopes afakwa inzalo ngemva kokusetshenziswa ngakunye ngabantu (Fig. 8).Izinqubo zokuhlanza zenziwe ngaphansi kokuqondisa nokugunyazwa koMnyango Wokulawula Amagciwane kanye Nezifo Eziwumshayabhuqe we-Michigan Medical Center kanye Nophiko Olumaphakathi Lokucutshungulwa Kwenzalo. Ngaphambi kocwaningo, amathuluzi ahlolelwa futhi aqinisekiswa ukuvala inzalo yi-Advanced Sterilization Products (ASP, Johnson & Johnson), ibhizinisi elihlinzeka ngezinsizakalo zokuvimbela ukutheleleka kanye nokuqinisekisa ukuvala inzalo. Ngaphambi kocwaningo, amathuluzi ahlolelwa futhi aqinisekiswa ukuvala inzalo yi-Advanced Sterilization Products (ASP, Johnson & Johnson), ibhizinisi elihlinzeka ngezinsizakalo zokuvimbela ukutheleleka kanye nokuqinisekisa ukuvala inzalo. Перед исследованием инструменты были протестированы и одобрены для стерилизации компанией Advanced Sterilization Products (ASP, Johnson & Johnson), коммерческой ордгайской, kanye ne-профилактике инфекций ne-проверке стерилизации. Ngaphambi kocwaningo, amathuluzi ahlolelwa futhi agunyazwa ukuvalwa inzalo yi-Advanced Sterilization Products (ASP, Johnson & Johnson), inhlangano yezohwebo ehlinzeka ngezinsizakalo zokuvimbela ukutheleleka kanye nokuqinisekisa ukuvala inzalo. Перед исследованием инструменты были стерилизованы и проверены Advanced Sterilization Products (ASP, Johnson & Johnson), коммерческой организацией, которая предокестакестанская й и проверке стерилизации. Amathuluzi asetshenziswa amagciwane futhi ahlolwa ngaphambi kocwaningo yi-Advanced Sterilization Products (ASP, Johnson & Johnson), inhlangano yezohwebo ehlinzeka ngezinsizakalo zokuvimbela ukutheleleka kanye nokuqinisekisa ukuvala inzalo.
Ithuluzi kabusha.(a) Ama-Endoscopes afakwa kumathreyi ngemva kokuvala inzalo ngakunye kusetshenziswa inqubo yokucubungula ye-STERRAD.(b) I-SMF nezintambo zinqanyulwa ngezixhumi ze-fiber optic nezikagesi, ngokulandelana kwazo, ezivalwa ngaphambi kokuthi zicutshungulwe kabusha.
Hlanza ama-endoscope ngokwenza lokhu okulandelayo: (1) sula i-endoscope ngendwangu engenantambo efakwe kumshini wokuhlanza i-enzymatic kusuka e-proximal kuya kwe-distal;(2) Gxilisa ithuluzi kusisombululo se-enzymatic detergent imizuzu emi-3 ngamanzi.indwangu engenalutho.Izixhumi zikagesi kanye ne-fiber optic zimbozwe futhi zikhishwe kwisixazululo;(3) I-endoscope iyagoqwa bese ifakwa kuthreyi yensimbi ukuze kubulawe inzalo kusetshenziswa i-STERRAD 100NX, i-hydrogen peroxide gas plasma.izinga lokushisa eliphansi uma kuqhathaniswa nendawo enomswakama ophansi.
Amasethi edatha asetshenzisiwe kanye/noma ahlaziywa ocwaningweni lwamanje ayatholakala kubabhali abafanele uma kunesicelo esifanele.
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