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Confocal laser endoscopy yog ib txoj hauv kev tshiab ntawm kev kho qhov muag ntawm lub sijhawm.Fluorescent dluab ntawm histological zoo tuaj yeem tau txais tam sim ntawd los ntawm lub epithelium ntawm hollow lub cev.Tam sim no, kev tshuaj ntsuam xyuas tau ua nyob ze nrog cov cuab yeej sojntsuam uas feem ntau siv hauv kev kho mob, nrog kev txwv tsis pub muaj kev tswj xyuas kom pom tseeb.Peb ua qauv qhia kev siv cov parametric resonant scanner mounted ntawm lub distal kawg ntawm ib qho endoscope ua high-speed lateral deflection.Ib lub qhov tau raug etched rau hauv nruab nrab ntawm lub reflector yob li txoj kev teeb.Qhov kev tsim no txo ​​qhov loj ntawm qhov ntsuas mus rau 2.4 mm inch thiab 10 mm ntev, tso cai rau nws mus rau pem hauv ntej los ntawm kev ua haujlwm ntawm cov qauv kho mob endoscopes.Lub lens compact muab lateral thiab axial daws teeb meem ntawm 1.1 thiab 13.6 µm, feem.Kev ua haujlwm nrug ntawm 0 µm thiab thaj chaw saib ntawm 250 µm × 250 µm tau ua tiav ntawm tus lej nce txog 20 Hz.Excitation ntawm 488 nm excites fluorescein, FDA pom zoo dye rau cov ntaub so ntswg zoo sib xws.Endoscopes tau rov ua dua rau 18 lub voj voog yam tsis muaj kev ua tsis tiav siv cov txheej txheem kho mob kom tsis muaj menyuam.Cov duab fluorescent tau txais los ntawm ib txwm colonic mucosa, tubular adenomas, hyperplastic polyps, ulcerative colitis, thiab Crohn's colitis thaum niaj hnub colonoscopy.Tib lub hlwb tuaj yeem txheeb xyuas, suav nrog cov colonocytes, goblet hlwb, thiab cov hlwb ua mob.Mucosal nta xws li cov qauv crypt, crypt kab noj hniav, thiab lamina propria tuaj yeem paub qhov txawv.Cov cuab yeej siv tau los ua ib qho ntxiv rau cov pa endoscopy.
Confocal laser endoscopy yog ib qho tshiab imaging modality yog tsim los rau kev kho mob siv raws li ib tug adjunct rau niaj hnub endoscopy1,2,3.Cov cuab yeej hloov tau yooj yim, fiber ntau-optic txuas tuaj yeem siv los txheeb xyuas cov kab mob hauv cov kab mob epithelial uas kab cov kab mob hauv nruab nrog cev, xws li cov nyuv.Cov txheej nyias nyias ntawm cov ntaub so ntswg yog cov metabolically nquag nquag thiab yog lub hauv paus ntawm ntau yam kab mob xws li mob qog noj ntshav, kab mob, thiab mob.Endoscopy tuaj yeem ua tiav subcellular daws teeb meem, muab lub sijhawm tiag tiag, ze-histological zoo hauv vivo cov duab los pab cov kws kho mob txiav txim siab kho mob.Lub cev nqaij daim tawv nqaij biopsy ua rau muaj kev pheej hmoo los ntshav thiab perforation.Ntau dhau los yog tsawg dhau ntawm cov qauv biopsy feem ntau tau sau.Txhua tus qauv tshem tawm ua rau tus nqi phais.Nws yuav siv sij hawm ob peb hnub rau cov qauv los soj ntsuam los ntawm tus kws kho mob.Thaum lub sijhawm tos cov txiaj ntsig pathology, cov neeg mob feem ntau muaj kev ntxhov siab.Nyob rau hauv sib piv, lwm yam kev kho mob imaging modalities xws li MRI, CT, PET, SPECT, thiab ultrasound tsis muaj lub spatial daws teeb meem thiab lub cev ceev yuav tsum tau mus pom cov txheej txheem epithelial nyob rau hauv vivo nrog real-time, subcellular daws teeb meem.
Kev sojntsuam raws li ntsuas (Cellvizio) yog tam sim no siv nyob rau hauv cov chaw kho mob los ua "optical biopsy".Tus qauv tsim yog raws li kev sib txuas ntawm fiber ntau optic bundle4 uas sau thiab xa cov duab fluorescent.Cov tub ntxhais fiber ntau ua haujlwm ua "lub qhov" kom spatial lim defocused lub teeb rau subcellular daws teeb meem.Kev tshuaj ntsuam xyuas yog ua nyob ze siv lub loj, loj galvanometer.Cov kev cai no txwv tsis pub muaj peev xwm ntawm cov cuab yeej tswj kom pom tseeb.Kev ua kom zoo ntawm cov kab mob epithelial thaum ntxov yuav tsum pom qhov pom hauv qab ntawm cov ntaub so ntswg los soj ntsuam kev ntxeem tau thiab txiav txim siab txoj kev kho kom tsim nyog.Fluorescein, tus neeg saib xyuas qhov sib txawv uas tau pom zoo los ntawm FDA, yog muab tso rau hauv cov hlab ntsha los qhia txog cov yam ntxwv ntawm cov epithelium. Cov endomicroscopes no muaj qhov ntev <2.4 hli nyob rau hauv txoj kab uas hla, thiab yuav kis tau yooj yim mus rau pem hauv ntej los ntawm biopsy channel ntawm standard kho mob endoscopes. Cov endomicroscopes no muaj qhov ntev <2.4 hli nyob rau hauv txoj kab uas hla, thiab yuav kis tau yooj yim mus rau pem hauv ntej los ntawm biopsy channel ntawm standard kho mob endoscopes. Эти эндомикроскопы имеют размеры <2,4 мм в диаметре и могут быть легко проведены через биопкасийный thiab эндоскопов. Cov endomicroscopes no yog <2.4 hli nyob rau hauv lub cheeb thiab tuaj yeem yooj yim dhau los ntawm biopsy channel ntawm tus qauv kho mob endoscopes.Cov borescopes no tsawg dua 2.4 hli nyob rau hauv txoj kab uas hla thiab yooj yim dhau los ntawm biopsy channel ntawm tus qauv kho mob borescopes.Qhov kev hloov pauv no tso cai rau ntau yam kev siv tshuaj kho mob thiab yog ywj pheej ntawm cov tuam txhab endoscope.Ntau qhov kev tshawb fawb soj ntsuam tau ua tiav siv cov cuab yeej ntsuas no, suav nrog kev tshawb pom ntxov ntawm cov qog nqaij hlav ntawm txoj hlab pas, plab, hnyuv, thiab qhov ncauj kab noj hniav.Cov txheej txheem kev yees duab tau tsim thiab kev nyab xeeb ntawm cov txheej txheem tau tsim.
Microelectromechanical systems (MEMS) yog lub tshuab muaj zog rau kev tsim thiab tsim cov tshuab luam ntawv me me uas siv nyob rau hauv qhov kawg ntawm endoscopes.Txoj hauj lwm no (kwv yees rau proximal) tso cai rau kev hloov pauv ntau dua hauv kev tswj hwm txoj haujlwm ua kom pom tseeb5,6.Ntxiv nrog rau qhov deflection lateral, lub distal mechanism kuj tseem tuaj yeem ua axial scans, tom qab lub hom phiaj scans, thiab random access scans.Cov peev txheej no ua rau muaj kev sib tham ntau dua ntawm cov xovtooj ntawm tes, suav nrog cov duab sib dhos ntsug 7, thaj chaw loj ntawm kev pom (FOV) 8 aberration-free scanning, thiab txhim kho kev ua tau zoo hauv cov neeg siv kev txiav txim siab sub-regions9.MEMS daws qhov teeb meem loj ntawm kev ntim cov tshuab luam ntawv nrog qhov chaw txwv tsis pub muaj nyob rau ntawm qhov kawg ntawm qhov ntsuas.Piv rau cov galvanometers loj, MEMS muab kev ua tau zoo tshaj ntawm qhov me me, kev kub ceev, thiab kev siv hluav taws xob tsawg.Cov txheej txheem tsim khoom yooj yim tuaj yeem ua kom tiav rau kev tsim khoom loj ntawm tus nqi qis.Ntau tus MEMS tsim tau tshaj tawm yav dhau los 10,11,12.Tsis muaj ib qho ntawm cov thev naus laus zis tseem tsis tau tsim txaus los ua kom muaj kev siv tshuaj kho mob dav dav ntawm lub sijhawm tiag tiag hauv vivo imaging los ntawm kev ua haujlwm ntawm kev kho mob endoscope.Ntawm no, peb tsom los ua kom pom kev siv MEMS scanner ntawm qhov kawg ntawm qhov endoscope rau hauv vivo tib neeg cov duab tau txais thaum lub sij hawm niaj hnub kho mob endoscopy.
Ib qho khoom siv fiber ntau tau tsim los siv MEMS scanner ntawm qhov kawg ntawm qhov kawg kom sau lub sijhawm tiag tiag hauv vivo fluorescent dluab nrog cov yam ntxwv zoo li histological.Ib hom fiber ntau (SMF) yog nyob rau hauv ib lub raj hloov tau yooj yim thiab zoo siab ntawm λex = 488 nm.Qhov kev teeb tsa no ua kom luv luv qhov ntev ntawm qhov kawg ntawm qhov kawg thiab tso cai rau nws mus rau pem hauv ntej los ntawm kev ua haujlwm ntawm cov qauv kho mob endoscopes.Siv cov lus taw qhia rau nruab nrab ntawm qhov muag pom.Cov lo ntsiab muag no yog tsim los ua kom tiav ze li ntawm diffractive axial daws teeb meem nrog tus lej aperture (NA) = 0.41 thiab ua haujlwm nrug = 0 µm13.Precision shims yog tsim los ua kom pom tseeb ntawm optics 14. Lub tshuab scanner tau ntim rau hauv ib qho endoscope nrog lub ntsej muag nruj 2.4 hli hauv txoj kab uas hla thiab 10 hli ntev (Fig. 1a).Cov qhov ntev no tso cai rau nws siv rau hauv kev kho mob raws li ib qho khoom siv thaum lub sij hawm endoscopy (Fig. 1b).Lub zog siab tshaj plaws ntawm qhov teeb meem laser ntawm cov ntaub so ntswg yog 2 mW.
Confocal laser endoscopy (CLE) thiab MEMS scanners.Daim duab qhia (a) cov cuab yeej ntim khoom nrog qhov ntev ntawm qhov nruab nrab ntawm qhov ntev ntawm 2.4 hli txoj kab uas hla thiab 10 hli ntev thiab (b) txoj kab ncaj nraim los ntawm kev ua haujlwm ntawm tus qauv kho mob endoscope (Olympus CF-HQ190L).(c) Saib pem hauv ntej ntawm lub scanner uas qhia txog lub teeb pom kev nrog lub nruab nrab aperture ntawm 50 µm los ntawm cov kab hluav taws xob excitation dhau.Lub scanner yog mounted rau ntawm ib tug gimbal tsav los ntawm ib txheej ntawm quadrature comb drives.Lub resonant zaus ntawm lub cuab yeej yog txiav txim los ntawm qhov loj ntawm lub caij nplooj ntoos hlav torsion.(d) Sab saib ntawm lub scanner uas qhia lub scanner mounted ntawm ib tug sawv nrog cov xov hlau txuas nrog electrode anchors uas muab cov ntsiab lus txuas rau tsav thiab lub zog teeb liab.
Lub tshuab scanning muaj ib lub gimbal-mounted reflector uas tau tsav los ntawm ib txheej ntawm comb-driven quadrature actuators deflect lub beam tom qab (XY dav hlau) nyob rau hauv ib tug Lissajous qauv (Fig. 1c).Ib lub qhov 50 µm nyob rau hauv txoj kab uas hla tau etched nyob rau hauv qhov chaw uas lub excitation beam dhau.Lub scanner yog tsav ntawm lub resonant zaus ntawm tus tsim, uas tuaj yeem hloov kho qhov ntev ntawm lub caij nplooj ntoos hlav torsion.Electrode anchors tau kos rau ntawm lub periphery ntawm lub cuab yeej los muab cov ntsiab lus txuas rau lub zog thiab tswj cov cim (Daim duab 1d).
Cov duab kos duab yog mounted rau ntawm lub tsheb thauj khoom uas tuaj yeem dov mus rau hauv chav ua haujlwm.Lub graphical user interface tau tsim los txhawb cov neeg siv nrog kev paub tsawg kawg nkaus, xws li kws kho mob thiab kws saib xyuas neeg mob.Manually xyuas lub scanner tsav zaus, beamform hom, thiab duab FOV.
Tag nrho qhov ntev ntawm qhov endoscope yog kwv yees li 4m kom tso cai rau tag nrho cov khoom siv los ntawm kev ua haujlwm ntawm tus qauv kho mob endoscope (1.68m), nrog rau qhov ntev ntxiv rau maneuverability.Thaum kawg ntawm qhov kawg ntawm qhov endoscope, SMF thiab cov xov hlau xaus hauv cov khoom sib txuas uas txuas mus rau fiber ntau optic thiab wired ports ntawm lub hauv paus chaw nres tsheb.Qhov kev teeb tsa muaj lub laser, lub lim dej, lub tshuab hluav taws xob siab thiab lub ntsuas hluav taws xob photomultiplier (PMT).Lub amplifier muab lub zog thiab tsav cov cim rau lub scanner.Lub tshuab lim dej ua ke ua ke lub laser excitation mus rau SMF thiab hla cov fluorescence rau PMT.
Endoscopes tau rov ua dua tom qab txhua txoj kev kho mob uas siv cov txheej txheem STERRAD kom tsis muaj menyuam thiab tuaj yeem tiv taus mus txog 18 lub voj voog yam tsis ua haujlwm.Rau cov tshuaj OPA, tsis muaj cov tsos mob ntawm kev puas tsuaj tom qab ntau tshaj 10 lub voj voog tua kab mob.OPA cov txiaj ntsig tau ua tau zoo dua STERRAD's, qhia tias lub neej ntawm endoscopes tuaj yeem txuas ntxiv los ntawm kev tua kab mob siab ntau dua li rov ua kom tsis muaj menyuam.
Cov duab daws teeb meem tau txiav txim siab los ntawm qhov taw tes kis ua haujlwm siv cov hlaws fluorescent nrog txoj kab uas hla ntawm 0.1 μm.Rau lateral thiab axial daws teeb meem, tag nrho qhov dav ntawm ib nrab siab tshaj plaws (FWHM) ntawm 1.1 thiab 13.6 µm, ntsig txog, tau ntsuas (Fig. 2a, b).
Cov duab xaiv.Lub lateral (a) thiab axial (b) kev daws teeb meem ntawm qhov tsom teeb pom kev zoo yog cov yam ntxwv los ntawm qhov taw qhia kis muaj nuj nqi (PSF) ntsuas siv fluorescent microspheres nrog txoj kab uas hla ntawm 0.1 μm.Qhov ntsuas qhov dav ntawm ib nrab siab tshaj plaws (FWHM) yog 1.1 thiab 13.6 µm, feem.Inset: nthuav dav pom ntawm ib qho microsphere nyob rau hauv qhov kev hloov pauv (XY) thiab axial (XZ) cov lus qhia tau qhia.(c) Cov duab fluorescent tau los ntawm tus qauv (USAF 1951) lub hom phiaj strip (liab oval) qhia tias pawg 7-6 tuaj yeem daws tau meej meej.(d) Duab ntawm 10 µm txoj kab uas hla dispersed fluorescent microspheres uas qhia ib daim duab teb saib ntawm 250 µm × 250 µm.PSFs hauv (a, b) tau tsim siv MATLAB R2019a (https://www.mathworks.com/).(c, d) Cov duab fluorescent tau sau los ntawm LabVIEW 2021 (https://www.ni.com/).
Cov duab fluorescent los ntawm cov qauv daws teeb meem lo ntsiab muag kom meej meej qhov sib txawv ntawm cov kab hauv pawg 7-6, uas tswj cov kev daws teeb meem siab sab nraud (Fig. 2c).Qhov chaw saib (FOV) ntawm 250 µm × 250 µm tau txiav txim siab los ntawm cov duab ntawm 10 µm txoj kab uas hla fluorescent hlaws dispersed ntawm cov ntaub npog (Fig. 2d).
Ib txoj hauv kev zoo rau PMT nce kev tswj hwm thiab kho theem yog siv nyob rau hauv kev tshuaj ntsuam xyuas qhov system kom txo tau cov khoom txav los ntawm endoscopes, colon peristalsis, thiab cov neeg mob ua pa.Cov duab rov tsim kho thiab ua cov algorithms tau piav qhia yav dhau los 14,15.PMT nce yog tswj los ntawm ib qho proportional-integral (PI) maub los tiv thaiv kev siv saturation16.Lub kaw lus nyeem qhov siab tshaj plaws pixel siv rau txhua lub thav duab, suav cov lus teb sib npaug thiab ib qho, thiab txiav txim siab PMT qhov txiaj ntsig kom ntseeg tau tias qhov kev siv pixel nyob rau hauv qhov kev tso cai.
Thaum nyob rau hauv vivo imaging, theem tsis sib haum ntawm scanner txav thiab tswj teeb liab tuaj yeem ua rau duab plooj.Cov teebmeem zoo li no tuaj yeem tshwm sim vim qhov hloov pauv ntawm qhov kub ntawm cov cuab yeej hauv tib neeg lub cev.Dawb lub teeb duab pom tau hais tias lub endoscope nyob rau hauv kev sib cuag nrog ib txwm colonic mucosa nyob rau hauv vivo (Daim duab 3a).Blurring ntawm misaligned pixels tuaj yeem pom hauv cov duab nyoos ntawm ib txwm colonic mucosa (Daim duab 3b).Tom qab kev kho mob nrog cov theem kom zoo thiab kev sib piv, cov yam ntxwv ntawm cov kab mob subcellular tuaj yeem paub qhov txawv (Fig. 3c).Yog xav paub ntxiv, cov duab nyoos confocal thiab cov duab ua tiav ntawm lub sijhawm muaj nyob rau hauv daim duab S1, thiab cov duab reconstruction tsis siv rau lub sijhawm tiag tiag thiab tom qab ua tiav tau nthuav tawm hauv Table S1 thiab Table S2.
Kev ua duab.(a) Wide-angle endoscopic duab uas qhia qhov endoscope (E) muab tso rau hauv kev sib cuag nrog ib txwm (N) colonic mucosa los sau hauv vivo fluorescent dluab tom qab fluorescein tswj.(b) Wander nyob rau hauv X thiab Y axes thaum lub sij hawm scanning yuav ua rau misaligned pixels plooj.Rau kev ua qauv qhia, kev hloov pauv loj yog siv rau cov duab qub.(c) Tom qab kho cov theem tom qab ua tiav, cov ntsiab lus ntawm cov mucosal tuaj yeem raug soj ntsuam, suav nrog cov qauv crypt (cov xub), nrog lub hauv paus lumen (l) ncig los ntawm lamina propria (lp).Tib lub hlwb tuaj yeem paub qhov txawv, suav nrog colonocytes (c), goblet cells (g), thiab inflammatory cells ( xub).Saib ntxiv video 1. (b, c) Cov duab ua tiav siv LabVIEW 2021.
Confocal fluorescence dluab tau txais nyob rau hauv vivo nyob rau hauv ntau yam kab mob colonic los ua kom pom qhov dav siv nyob rau hauv lub cuab yeej.Wide-angle imaging yog thawj zaug uas siv lub teeb dawb kom pom cov mucosa txawv txav.Lub endoscope tom qab ntawd tau nce siab los ntawm txoj haujlwm ua haujlwm ntawm colonoscope thiab coj mus rau hauv kev sib cuag nrog cov mucosa.
Wide-field endoscopy, confocal endomicroscopy, thiab histology (H&E) dluab yog qhia rau colonic neoplasia, nrog rau tubular adenoma thiab hyperplastic polyp. Wide-field endoscopy, confocal endomicroscopy, thiab histology (H&E) dluab yog qhia rau colonic neoplasia, nrog rau tubular adenoma thiab hyperplastic polyp. Широкопольная эндоскопия, конфокальная эндомикроскопия и гистологические (H&E) изображения покалятаны ки, включая тубулярную аденому и гиперпластический полип. Colonic endoscopy, confocal endomicroscopy, thiab histological (H&E) imaging yog qhia rau colonic neoplasia, nrog rau tubular adenoma thiab hyperplastic polyp.在显示结肠肿瘤（包括管状腺瘤和增生性息肉）的广角内窥镜检、共聚焦显微内窻囇像.共设计脚肠化(图像管状躰化和增生性息肉)的广角内刵霱录共共共八在微禷全duab. Широкопольная эндоскопия, конфокальная микроэндоскопия и гистологические (H&E) изображения, покатокопие в покатологические thiab, включая тубулярные аденомы и гиперпластические полипы. Broad-field endoscopy, confocal microendoscopy, thiab histological (H&E) cov duab qhia cov qog ntawm txoj hnyuv, suav nrog tubular adenomas thiab hyperplastic polyps.Tubular adenomas pom qhov poob ntawm ib txwm crypt architecture, txo qhov loj ntawm goblet hlwb, distortion ntawm crypt lumen, thiab thickening ntawm lamina propria (Fig. 4a-c).Hyperplastic polyps pom stellate architecture ntawm crypts, ob peb lub goblet hlwb, slit-zoo li lumen ntawm crypts, thiab tsis zoo lamellar crypts (Fig. 4d-f).
Daim duab ntawm daim tawv nqaij tuab hauv vivo. Cov neeg sawv cev dawb lub teeb endoscopy, confocal endomicroscope, thiab histology (H&E) cov duab tau qhia rau (ac) adenoma, (df) hyperplastic polyp, (gi) ulcerative colitis, thiab (jl) Crohn's colitis. Cov neeg sawv cev dawb lub teeb endoscopy, confocal endomicroscope, thiab histology (H&E) cov duab tau qhia rau (ac) adenoma, (df) hyperplastic polyp, (gi) ulcerative colitis, thiab (jl) Crohn's colitis. Типичные изображения эндоскопии в белом свете, конфокального эндомикроскопа и гимстологии (H&E) по дата (H&E) иперпластического полипа, (gi) язвенного колита и (jl) колита Крона. Cov duab dawb-light endoscopy, confocal endomicroscope, thiab histology (H&E) cov duab tau qhia rau (ac) adenoma, (df) hyperplastic polyp, (gi) ulcerative colitis, thiab (jl) Crohn's colitis.显示了(ac) 腺瘤、(df) 增生性息肉、(gi) 溃疡性结肠炎和(jl) 克罗恩结肠炎的代表怀焦内窥镜检 在和组织学( H&E) 图像. Nws qhia (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炅充肠肠炎性和电视学 (H&E) ) duab. Представлены репрезентативные эндоскопия в белом свете, конфокальная эндоскопия и гистология (ac) (ac) кого полипоза, (gi) язвенного колита и (jl) колита Крона (H&E). Cov neeg sawv cev dawb-lub teeb endoscopy, confocal endoscopy, thiab histology ntawm (ac) adenoma, (df) hyperplastic polyposis, (gi) ulcerative colitis, thiab (jl) Crohn's colitis (H&E) tau qhia.(B) qhia cov duab confocal tau hauv vivo los ntawm tubular adenoma (TA) siv qhov endoscope (E).Cov kab mob precancerous no qhia tau tias poob ntawm ib txwm crypt architecture (xub), distortion ntawm lub crypt lumen (l), thiab cov neeg coob coob ntawm crypt lamina propria (lp).Colonocytes (c), goblet cells (g), thiab inflammatory cells ( xub) kuj raug txheeb xyuas.Smt.Ntxiv Video 2. (e) qhia cov duab confocal tau los ntawm ib tug hyperplastic polyp (HP) nyob rau hauv vivo.Qhov mob hnyav no ua rau pom lub ntsej muag crypt architecture (xub), ib qho zoo li crypt lumen (l), thiab ib qho tsis xwm yeem zoo li lamina propria (lp).Colonocytes (c), ob peb goblet hlwb (g) thiab inflammatory cells ( xub) kuj raug txheeb xyuas.Smt.Ntxiv Video 3. (h) qhia cov duab confocal tau txais hauv cov kab mob ulcerative colitis (UC) hauv vivo.Cov kab mob inflammatory no qhia tau hais tias distorted crypt architecture ( xub) thiab tseem ceeb goblet hlwb (g).Feathers ntawm fluorescein (f) yog extruded los ntawm epithelial hlwb, xav txog kev nce vascular permeability.Ntau cov kab mob inflammatory (cov xub) pom nyob rau hauv lamina propria (lp).Smt.Ntxiv Video 4. (k) qhia ib daim duab confocal tau nyob rau hauv vivo los ntawm ib cheeb tsam ntawm Crohn's colitis (CC).Cov kab mob inflammatory no qhia tau hais tias distorted crypt architecture ( xub) thiab tseem ceeb goblet hlwb (g).Feathers ntawm fluorescein (f) yog extruded los ntawm epithelial hlwb, xav txog kev nce vascular permeability.Ntau cov kab mob inflammatory (cov xub) pom nyob rau hauv lamina propria (lp).Smt.Ntxiv Video 5. (b, d, h, l) Cov duab ua tiav siv LabVIEW 2021.
Cov duab zoo sib xws ntawm cov kab mob colonic tau pom, suav nrog ulcerative colitis (UC) (Daim duab 4g-i) thiab Crohn's colitis (Daim duab 4j-l).Cov lus teb inflammatory yog xav tias yog tus cwj pwm los ntawm cov txheej txheem crypted uas muaj cov kab mob goblet protruding.Fluorescein yog nyem tawm ntawm cov hlwb epithelial, xav txog kev nce vascular permeability.Cov kab mob inflammatory ntau tuaj yeem pom hauv lamina propria.
Peb tau ua kom pom qhov kev kho mob ntawm cov khoom siv fiber ntau-coupled confocal laser endoscope uas siv lub distally positioned MEMS scanner rau hauv vivo duab tau.Ntawm resonant zaus, ncej tus nqi mus txog 20 Hz tuaj yeem ua tiav siv Lissajous scan hom siab kom txo cov khoom cuav.Txoj kev kho qhov muag yog folded los muab cov nqaj nthuav dav thiab cov lej aperture txaus kom ua tiav qhov kev daws teeb meem sab nraud ntawm 1.1 µm.Fluorescent dluab ntawm histological zoo tau txais thaum lub sij hawm niaj hnub colonoscopy ntawm ib txwm colonic mucosa, tubular adenomas, hyperplastic polyps, ulcerative colitis, thiab Crohn's colitis.Tib lub hlwb tuaj yeem txheeb xyuas, suav nrog cov colonocytes, goblet hlwb, thiab cov hlwb ua mob.Mucosal nta xws li cov qauv crypt, crypt kab noj hniav, thiab lamina propria tuaj yeem paub qhov txawv.Cov khoom siv precision yog micro-machined los xyuas kom meej cov kev sib raug zoo ntawm tus neeg kho qhov muag thiab cov khoom siv kho qhov muag hauv 2.4mm txoj kab uas hla x 10mm ntev ntsuas.Kev tsim kho qhov muag txo qhov ntev ntawm lub qhov muag tsis pom kev txaus kom tso cai ncaj qha los ntawm tus qauv loj (3.2 hli txoj kab uas hla) ua haujlwm hauv kev kho mob endoscopes.Yog li ntawd, tsis hais cov chaw tsim khoom, cov cuab yeej siv tau dav siv los ntawm cov kws kho mob ntawm qhov chaw nyob.Excitation tau ua ntawm λex = 488 nm kom zoo siab fluorescein, FDA pom zoo dye, kom tau txais qhov sib txawv siab.Cov cuab yeej tau rov ua dua yam tsis muaj teeb meem rau 18 lub voj voog uas siv cov txheej txheem kho kom tsis muaj menyuam.
Ob qho lwm yam kev ntsuas ntsuas tau raug kuaj xyuas.Cellvizio (Mauna Kea Technologies) yog ib qho kev sojntsuam-raws li confocal laser endoscope (pCLE) uas siv ib pob ntawm multimode coherent fiber optic cables los sau thiab xa cov duab fluorescence1.Ib daim iav galvo nyob rau ntawm lub hauv paus chaw nres tsheb ua ib qho kev soj ntsuam lateral ntawm qhov kawg.Optical seem yog sau rau hauv kab rov tav (XY) dav hlau nrog qhov tob ntawm 0 txog 70 µm.Cov khoom siv microprobe muaj los ntawm 0.91 (19 G koob) mus rau 5 hli inch.Kev daws teeb meem sab nraud ntawm 1 txog 3.5 µm tau ua tiav.Cov duab tau sau los ntawm tus ncej tus nqi ntawm 9 txog 12 Hz nrog rau ib qho kev pom ntawm 240 txog 600 µm.Lub platform tau siv nyob rau hauv ntau qhov chaw xws li cov kav dej, lub zais zis, txoj hnyuv, txoj hlab pas, ntsws, thiab pancreas.Optiscan Pty Ltd tau tsim ib qho endoscope-raws li confocal laser endoscope (eCLE) nrog lub tshuab scanning ua rau hauv lub raj ntxig (distal kawg) ntawm tus kws tshaj lij endoscope (EC-3870K, Pentax Precision Instruments) 17 .Qhov chaw kho qhov muag tau ua tiav siv ib hom fiber ntau, thiab sab scanning yog nqa tawm siv lub cantilever mechanism los ntawm lub resonant tuning diav rawg.Ib Daim Duab Memory Alloy (Nitinol) actuator yog siv los tsim axial hloov chaw.Tag nrho txoj kab uas hla ntawm confocal module yog 5 mm.Rau kev tsom xam, GRIN lens nrog tus lej aperture ntawm NA = 0.6 yog siv.Kab rov tav dluab tau txais nrog lateral thiab axial kev daws teeb meem ntawm 0.7 thiab 7 µm, ntsig txog, ntawm tus ncej tus nqi ntawm 0.8-1.6 Hz thiab thaj tsam ntawm 500 µm × 500 µm.
Peb ua qauv qhia kev daws teeb meem hauv vivo fluorescence imaging tau los ntawm tib neeg lub cev los ntawm kev kho mob endoscope siv lub distal kawg MEMS scanner.Fluorescence muab cov duab zoo sib xws, thiab ligands uas khi rau lub hom phiaj ntawm tes tuaj yeem sau nrog fluorophores los muab cov cim molecular rau kev txhim kho kab mob.Lwm cov txheej txheem kho qhov muag rau hauv vivo microendoscopy kuj tau tsim. OCT siv qhov luv luv coherence ntev los ntawm lub teeb pom kev dav dav los sau cov duab hauv lub dav hlau ntsug nrog qhov tob> 1 mm19. OCT siv qhov luv luv coherence ntev los ntawm lub teeb pom kev dav dav los sau cov duab hauv lub dav hlau ntsug nrog qhov tob> 1 mm19. ОКТ использует короткую длину когерентности широкополосного источника света для сбора изобркаатьрений hlубиной > 1 мм19. OCT siv qhov luv luv coherence ntev ntawm lub teeb pom kev dav dav kom tau txais cov duab hauv lub dav hlau ntsug nrog> 1 hli qhov tob 19. OCT 使用宽带光源的短相干长度来收集垂直平面中深度> 1 mm19 的图像.1mm19 ua. ОКТ использует короткую длину когерентности широкополосного источника света для сбора изобравжений на 1нгравений на ой плоскости. OCT siv qhov luv luv coherence ntev ntawm lub teeb pom kev dav dav kom tau txais cov duab> 1 mm19 hauv lub dav hlau ntsug.Txawm li cas los xij, qhov kev sib piv qis no yog nyob ntawm kev sau cov duab backscattered thiab cov duab daws teeb meem yog txwv los ntawm speckle artifacts.Photoacoustic endoscopy generates nyob rau hauv vivo dluab raws li sai sai thermoelastic expansion nyob rau hauv cov ntaub so ntswg tom qab nqus tau ib tug laser mem tes uas generates suab tsis 20. Txoj hauv kev no tau pom qhov pom qhov tob> 1 cm hauv tib neeg txoj hnyuv hauv vivo los saib xyuas kev kho mob. Txoj hauv kev no tau pom qhov pom qhov tob> 1 cm hauv tib neeg txoj hnyuv hauv vivo los saib xyuas kev kho mob. Этот подход продемонстрировал глубину визуализации > 1 см в толстой кишке человека in vivo для мониторинаптторинга. Txoj hauv kev no tau pom qhov pom qhov tob ntawm> 1 cm hauv tib neeg txoj hnyuv hauv vivo rau kev saib xyuas kev kho mob.这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.这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 мониторитринг. Txoj hauv kev no tau ua kom pom ntawm qhov tob tob> 1 cm hauv tib neeg txoj hnyuv hauv vivo los saib xyuas kev kho mob.Qhov sib txawv tsuas yog tsim los ntawm hemoglobin hauv vasculature.Multiphoton endoscopy generates high-contrast fluorescence dluab thaum ob los yog ntau tshaj NIR photons tsoo cov ntaub so ntswg biomolecules ib txhij21. Txoj hauv kev no tuaj yeem ua tiav cov duab pom qhov tob> 1 hli nrog cov duab tsis zoo. Txoj hauv kev no tuaj yeem ua tiav cov duab pom qhov tob> 1 hli nrog cov duab tsis zoo. Этот подход может обеспечить глубину изображения > 1 мм с низкой фототоксичностью. Txoj hauv kev no tuaj yeem muab cov duab qhov tob> 1 hli nrog qhov tsis tshua muaj phototoxicity.这种方法可以实现>1 毫米的成像深度，光毒性低.这种方法可以实现>1 毫米的成像深度，光毒性低. Этот подход может обеспечить глубину изображения > 1 мм с низкой фототоксичностью. Txoj hauv kev no tuaj yeem muab cov duab qhov tob> 1 hli nrog qhov tsis tshua muaj phototoxicity.Siab siv femtosecond laser pulses yuav tsum tau thiab cov qauv no tsis tau kuaj pom tseeb thaum lub sij hawm endoscopy.
Nyob rau hauv cov qauv no, lub scanner ua tsuas yog tom qab deflection, yog li qhov kho qhov muag yog nyob rau hauv kab rov tav (XY) dav hlau.Cov cuab yeej muaj peev xwm ua haujlwm ntawm tus ncej siab dua (20 Hz) dua li cov iav galvanic (12 Hz) hauv Cellvizio system.Ua kom tus ncej tus nqi kom txo tau cov duab kos thiab txo tus ncej tus nqi kom txhawb lub teeb liab.Kev kub ceev thiab kev siv lub tshuab hluav taws xob yuav tsum tau ua kom txo tau cov khoom siv hluav taws xob loj uas tshwm sim los ntawm kev ua haujlwm endoscopic, kev ua pa ua pa, thiab lub plab zom mov.Parametric resonant scanners tau pom tias ua tiav axial hloov pauv ntau dua li ntau pua microns22. Cov duab tuaj yeem sau rau hauv lub dav hlau ntsug (XZ), nyob rau sab laug rau ntawm qhov chaw mucosal, muab tib qhov kev pom zoo li ntawm histology (H&E). Cov duab tuaj yeem sau rau hauv lub dav hlau ntsug (XZ), nyob rau sab laug rau ntawm qhov chaw mucosal, muab tib qhov kev pom zoo li ntawm histology (H&E). Изображения могут быть получены в вертикальной плоскости (XZ), перпендикулярной поверхности слизистой ь такое же изображение, как при гистологии (H&E). Cov duab tuaj yeem muab coj los rau hauv lub dav hlau ntsug (XZ) perpendicular mus rau qhov chaw mucosal kom muab cov duab tib yam li hauv 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) 相君的。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 ь такое же изображение, как при гистологическом исследовании (H&E). Cov duab tuaj yeem muab coj los rau hauv lub dav hlau ntsug (XZ) perpendicular mus rau qhov chaw mucosal kom muab cov duab tib yam li kev kuaj mob histological (H&E).Lub tshuab luam ntawv tuaj yeem muab tso rau hauv txoj haujlwm tom qab lub hom phiaj qhov twg lub teeb pom kev zoo poob rau ntawm qhov chaw kho qhov muag tseem ceeb kom txo qis rhiab rau aberrations8.Ze li ntawm diffraction-limited focal ntim tuaj yeem sib txawv ntawm qhov chaw loj ntawm qhov pom.Random access scanning tuaj yeem ua rau deflect reflectors rau tus neeg siv txoj haujlwm 9.Lub teb ntawm kev pom tuaj yeem raug txo kom pom cov cheeb tsam tsis txaus ntseeg ntawm daim duab, txhim kho lub teeb liab-rau-suab nrov piv, qhov sib piv, thiab tus ncej tus nqi.Cov tshuab luam ntawv tuaj yeem ua tau loj-tsim siv cov txheej txheem yooj yim.Pua pua ntawm cov khoom siv tuaj yeem ua rau ntawm txhua lub silicon wafer kom nce ntau lawm rau cov nqi qis thiab kev xa tawm dav.
Txoj kev folded lub teeb yuav txo qhov luaj li cas ntawm lub qhov muag tsis pom kev, ua rau nws yooj yim siv lub endoscope ua ib qho khoom siv thaum lub sij hawm niaj hnub colonoscopy.Nyob rau hauv cov duab fluorescent pom, subcellular nta ntawm mucosa tuaj yeem pom qhov txawv ntawm tubular adenomas (precancerous) los ntawm hyperplastic polyps (benign).Cov txiaj ntsig no qhia tau tias kev tshuaj ntsuam xyuas endoscopy tuaj yeem txo tus naj npawb ntawm qhov tsis tsim nyog biopsies23.Cov teeb meem dav dav cuam tshuam nrog kev phais tuaj yeem txo qis, kev soj ntsuam lub sijhawm tuaj yeem ua kom zoo dua qub, thiab kev soj ntsuam histological ntawm cov kab mob me tuaj yeem txo qis.Peb kuj tseem qhia hauv vivo cov duab ntawm cov neeg mob uas muaj kab mob plab hnyuv, nrog rau cov kab mob plab hnyuv (UC) thiab Crohn's colitis.Pa dawb lub teeb colonoscopy muab macroscopic saib ntawm mucosal nto nrog txwv tsis pub muaj peev xwm los ntsuas qhov tseeb ntawm mucosal kho.Endoscopy tuaj yeem siv hauv vivo los ntsuas qhov ua tau zoo ntawm cov tshuaj lom neeg xws li tshuaj tiv thaiv TNF24.Qhov tseeb ntawm vivo kev ntsuam xyuas kuj tuaj yeem txo lossis tiv thaiv kab mob rov qab thiab teeb meem xws li kev phais thiab txhim kho lub neej zoo.Tsis muaj kev phiv loj heev tau tshaj tawm hauv kev tshawb fawb soj ntsuam cuam tshuam nrog kev siv cov tshuaj fluorescein-muaj endoscopes hauv vivo25. Lub zog laser ntawm lub mucosal nto tau txwv rau <2 mW kom txo qis kev pheej hmoo rau kev raug mob thermal thiab ua tau raws li FDA cov cai rau qhov tsis muaj kev pheej hmoo tseem ceeb26 rau 21 CFR 812. Lub zog laser ntawm lub mucosal nto tau txwv rau <2 mW kom txo qhov kev pheej hmoo ntawm kev raug mob thermal thiab ua tau raws li FDA cov cai rau qhov tsis tseem ceeb txaus ntshai26 rau 21 CFR 812. Мощность лазера на поверхности слизистой оболочки была ограничена до <2 мВт, чтобы свести к минимукним у thiab соответствовать требованиям FDA относительно незначительного риска26 согласно 21 CFR 812. Lub zog laser ntawm lub mucosal nto yog txwv rau <2 mW kom txo tau qhov kev pheej hmoo ntawm thermal puas thiab ua tau raws li FDA cov cai rau kev pheej hmoo tsis zoo26 hauv qab 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，并满足要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 度地降低热损伤风险。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 минимукним у thiab соответствовать требованиям FDA 21 CFR 812 относительно незначительного риска26. Lub zog laser ntawm lub mucosal nto tau txwv rau <2 mW kom txo tau qhov kev pheej hmoo ntawm thermal puas thiab ua tau raws li FDA 21 CFR 812 cov cai rau kev pheej hmoo tsis zoo26.
Kev tsim cov cuab yeej tuaj yeem hloov kho los txhim kho cov duab zoo.Tshwj xeeb optics muaj los txo cov kheej kheej aberration, txhim kho cov duab daws teeb meem thiab nce kev ua haujlwm nrug.SIL tuaj yeem hloov kho kom zoo dua qhov ntsuas qhov ntsuas ntawm cov ntaub so ntswg (~ 1.4) txhawm rau txhim kho lub teeb coupling.Tus tsav zaus tuaj yeem hloov kho kom nce lub kaum sab xis ntawm lub scanner thiab nthuav dav cov duab pom.Koj tuaj yeem siv txoj kev siv lub tshuab kom tshem tawm cov thav ntawv ntawm cov duab nrog kev txav tseem ceeb los txo cov nyhuv no.Lub teb-programmable rooj vag array (FPGA) nrog high-ceev cov ntaub ntawv tau txais yuav raug siv los muab kev ua tau zoo ntawm lub sijhawm ua haujlwm puv-ntev.Rau kev siv tshuaj kho mob ntau dua, cov txheej txheem automated yuav tsum kho rau theem hloov pauv thiab cov duab kos duab rau kev txhais cov duab tiag tiag.Lub monolithic 3-axis parametric resonant scanner tuaj yeem siv los qhia axial scanning 22 . Cov cuab yeej no tau tsim los kom ua tiav qhov tsis tau pom dua los ntawm txoj kab ntsug> 400 µm los ntawm kev kho cov tsav zaus hauv kev tswj hwm uas muaj kev sib xyaw ua kom softening / stiffening dynamics27. Cov cuab yeej no tau tsim los kom ua tiav qhov tsis tau pom dua los ntawm txoj kab ntsug> 400 µm los ntawm kev kho cov tsav zaus hauv kev tswj hwm uas muaj kev sib xyaw ua kom softening / stiffening dynamics27. Эти устройства были разработаны для достижения беспрецедентного вертикального смещения > 400 мкм на приникального смещения > 400 мкм на приникального kuv в режиме, который характеризуется смешанной динамикой смягчения/жесткости27. Cov khoom siv no tau tsim los ua kom tiav qhov tsis tau pom dua los ntawm txoj kab ntsug ntawm> 400 µm los ntawm kev teeb tsa lub zaus tsav nyob rau hauv hom uas yog cov yam ntxwv ntawm kev sib xyaw mos / tawv dynamics27.这些设备的开发是为了通过在具有混合软化/硬化动力学的状态下调整驱剪动频率来的状态下调整驱剪剪率来垂直位移27.这些设备的开发是为了在具有混合软化硬化学学的状态縋 调敇驱学的状态 下调敇驱小0 µm 的 垂直位移 27. Эти устройства были разработаны для достижения беспрецедентных вертикальных смещений > 400 мкм пырисата на в режиме со смешанной кинетикой размягчения/затвердевания27. Cov cuab yeej no tau tsim los kom ua tiav qhov tsis tau pom dua los ntawm txoj kab ntsug> 400 µm los ntawm kev kho qhov cuam tshuam zaus hauv kev sib xyaw softening / hardening kinetics hom27.Nyob rau hauv lub neej yav tom ntej, ntsug transverse imaging tej zaum yuav pab nyob rau hauv staging ntxov cancer (T1a).Lub capacitive sensing Circuit Court tuaj yeem siv los taug qab kev txav ntawm lub scanner thiab kho rau theem hloov 28.Tsis siv neeg theem calibration siv lub sensor Circuit Court hloov tau phau ntawv ntsuas ntsuas calibration ua ntej siv.Kev ntseeg tau ntawm cov cuab yeej tuaj yeem txhim kho los ntawm kev siv cov cuab yeej siv txhim khu kev qha ntau dua los ua kom cov txheej txheem ua tiav.MEMS thev naus laus zis tau cog lus tias yuav ua kom nrawm rau kev siv cov endoscopes rau kev pom cov epithelium ntawm cov kab mob hauv nruab nrog cev, kuaj kab mob, thiab saib xyuas kev kho mob hauv txoj hauv kev tsawg tsawg.Nrog rau kev txhim kho ntxiv, qhov kev hloov kho tshiab no tuaj yeem dhau los ua qhov kev daws teeb meem qis los siv los ua kev kho mob endoscopes rau kev kuaj histological tam sim ntawd thiab tuaj yeem hloov pauv cov tshuaj ntsuam xyuas kab mob.
Ray tracing simulations tau ua tiav siv ZEMAX optical tsim software (version 2013) los txiav txim qhov tsis muaj qhov tsom teeb pom kev zoo.Cov qauv tsim muaj xws li ze-diffractive axial daws teeb meem, ua haujlwm nrug = 0 µm, thiab thaj chaw saib (FOV) ntau dua 250 × 250 µm2.Rau excitation ntawm lub wavelength λex = 488 nm, ib hom fiber ntau (SMF) tau siv.Achromatic doublets yog siv los txo qhov sib txawv ntawm cov fluorescence sau (Daim duab 5a).Lub beam hla los ntawm SMF nrog ib hom teb txoj kab uas hla ntawm 3.5 μm thiab tsis muaj truncation dhau los ntawm qhov chaw ntawm lub reflector nrog ib tug aperture txoj kab uas hla ntawm 50 μm.Siv lub zog immersion (hemispherical) lo ntsiab muag nrog lub siab refractive index (n = 2.03) kom txo qhov teeb meem beam spherical aberration thiab xyuas kom meej tag nrho kev sib cuag nrog lub mucosal nto.Focusing optics muab tag nrho NA = 0.41, qhov twg NA = nsinα, n yog qhov cuam tshuam ntawm cov ntaub so ntswg, α yog lub siab tshaj plaws beam convergence kaum.Lub diffraction-limited lateral thiab axial daws teeb meem yog 0.44 thiab 6.65 µm, ntsig txog, siv NA = 0.41, λ = 488 nm, thiab n = 1.3313.Tsuas yog cov khoom lag luam muaj nyob nrog txoj kab uas hla sab nraud (OD) ≤ 2 mm raug txiav txim siab.Txoj kev kho qhov muag yog folded, thiab cov nqaj tawm ntawm SMF hla lub hauv paus aperture ntawm lub scanner thiab rov qab los ntawm daim iav ruaj khov (0.29 hli inch).Qhov kev teeb tsa no ua rau qhov ntev ntawm qhov kawg ntawm qhov kawg kom yooj yim rau kev nkag mus rau ntawm qhov endoscope los ntawm tus qauv (3.2 hli txoj kab uas hla) ua haujlwm ntawm kev kho mob endoscopes.Qhov no ua rau nws yooj yim siv los ua ib qho khoom siv thaum lub sij hawm niaj hnub endoscopy.
Folded light guide thiab endoscope ntim.(a) Lub nqaj excitation tawm ntawm OBC thiab dhau los ntawm lub hauv paus aperture ntawm lub scanner.Lub beam yog nthuav thiab xav txog los ntawm ib tug ruaj khov daim iav rov qab mus rau hauv lub scanner rau lateral deflection.Lub tsom optics muaj xws li ib khub ntawm achromatic doublet lo ntsiab muag thiab ib qho khoom immersion (hemispherical) lens muab kev sib cuag nrog lub mucosal nto.ZEMAX 2013 (https://www.zemax.com/) rau kev tsim kho qhov muag thiab cov kab hluav taws xob tracing simulation.(b) Qhia qhov chaw ntawm ntau yam khoom siv, suav nrog ib hom fiber ntau (SMF), scanner, iav, thiab lo ntsiab muag.Solidworks 2016 (https://www.solidworks.com/) tau siv rau 3D qauv ntawm lub ntim endoscope.
Ib qho SMF (# 460HP, Thorlabs) nrog lub hom phiaj txoj kab uas hla ntawm 3.5 µm ntawm lub wavelength ntawm 488 nm tau siv los ua "lub qhov" rau spatial filtering ntawm lub teeb defocused (Fig. 5b).Cov SMFs tau muab ntim rau hauv cov raj hloov pauv tau yooj yim (#Pebax 72D, Nordson MEDICAL).Qhov ntev ntawm kwv yees li 4 meters yog siv los xyuas kom muaj kev ncua deb ntawm tus neeg mob thiab cov duab kos duab.Ib khub ntawm 2 hli MgF2 coated achromatic doublet lo ntsiab muag (# 65568, # 65567, Edmund Optics) thiab 2 hli uncoated hemispherical lens (#90858, Edmund Optics) tau siv los tsom cov nqaj thiab sau cov fluorescence.Ntxig ib lub raj stainless hlau kawg (4 hli ntev, 2.0 hli OD, 1.6 hli ID) ntawm cov resin thiab lub raj sab nraud kom cais cov scanner vibration.Siv cov nplaum kho mob los tiv thaiv cov cuab yeej los ntawm cov kua hauv lub cev thiab cov txheej txheem tuav.Siv cov cua sov shrink tubing los tiv thaiv cov connectors.
Lub compact scanner yog tsim los ntawm lub hauv paus ntsiab lus ntawm parametric resonance.Etch a 50 µm aperture ntawm qhov chaw ntawm lub reflector kom xa cov kab tawm excitation.Siv cov txheej txheem quadrature comb-driven drives, cov nqaj nthuav dav yog deflected transversely hauv orthogonal kev taw qhia (XY dav hlau) hauv Lissajous hom.Lub rooj tsavxwm nrhiav tau cov ntaub ntawv (#DAQ PCI-6115, NI) tau siv los tsim cov cim analog los tswj lub scanner.Lub hwj chim tau muab los ntawm lub tshuab hluav taws xob siab (#PDm200, PiezoDrive) ntawm cov xov hlau nyias (#B4421241, MWS Wire Industries).Ua cov xaim ntawm cov electrode armature.Lub scanner ua haujlwm ntawm zaus ze li 15 kHz (ceev axis) thiab 4 kHz (slow axis) kom ua tiav FOV mus txog 250 µm × 250 µm.Yees duab tuaj yeem tua ntawm tus ncej tus nqi ntawm 10, 16, lossis 20 Hz.Cov thav duab no yog siv los ua kom phim tus nqi rov qab ntawm Lissajous scan qauv, uas nyob ntawm tus nqi ntawm X thiab Y excitation zaus ntawm lub scanner29.Cov ntsiab lus ntawm kev lag luam tawm ntawm tus ncej tus nqi, pixel daws teeb meem, thiab scan qauv ntom tau nthuav tawm hauv peb txoj haujlwm dhau los 14.
Lub zog lub xeev laser (#OBIS 488 LS, coherent) muab λex = 488 nm kom muaj zog fluorescein rau cov duab sib piv (Fig. 6a).Optical pigtails txuas nrog lub lim dej ntawm FC/APC connectors (poob 1.82 dB) (Fig. 6b).Lub nqaj yog deflected los ntawm daim iav dichroic (#WDM-12P-111-488/500:600, Oz Optics) hauv SMF los ntawm lwm lub FC / APC connector.Raws li 21 CFR 812, qhov xwm txheej lub zog rau cov ntaub so ntswg raug txwv rau qhov siab tshaj plaws ntawm 2 mW kom tau raws li FDA cov cai rau qhov tsis txaus ntseeg.Fluorescence tau dhau los ntawm daim iav dichroic thiab lub lim xa mus ntev (#BLP01-488R, Semrock).Fluorescence tau xa mus rau lub photomultiplier tube (PMT) ntes (#H7422-40, Hamamatsu) ntawm FC / PC connector siv lub ~ 1 m ntev multimode fiber ntau nrog 50 µm core inch.Cov teeb liab fluorescent tau nthuav dav nrog lub siab ceev tam sim no amplifier (#59-179, Edmund Optics).Cov software tshwj xeeb (LabVIEW 2021, NI) tau tsim los rau kev nrhiav cov ntaub ntawv ntawm lub sijhawm thiab ua cov duab.Lub zog laser thiab PMT nce nqis tau txiav txim siab los ntawm microcontroller (#Arduino UNO, Arduino) siv lub rooj tsav xwm tshwj xeeb luam tawm.Lub SMF thiab cov xov hlau xaus hauv cov khoom sib txuas thiab txuas mus rau fiber ntau optic (F) thiab wired (W) cov chaw nres nkoj ntawm lub hauv paus chaw nres tsheb (Daim duab 6c).Cov duab thaij duab muaj nyob rau ntawm lub laub nqa tau (Daim duab 6d). Ib qho kev sib cais transformer tau siv los txwv cov xau tam sim no rau <500 μA. Ib qho kev sib cais transformer tau siv los txwv cov xau tam sim no rau <500 μA. Для ограничения тока утечки до <500 мкА использовался изолирующий трансформатор. Ib qho kev sib cais transformer tau siv los txwv cov xau tam sim no rau <500 µA.使用隔离变压器将泄漏电流限制 nyob <500 μA. <500 μA. Используйте изолирующий трансформатор, чтобы ограничить ток утечки до <500 мкА. Siv lub tshuab hluav taws xob cais tawm kom txwv cov xau tam sim no mus rau <500µA.
kev pom qhov system.(a) PMT, laser thiab amplifier yog nyob rau hauv lub hauv paus chaw nres tsheb.(b) Hauv lub txhab nyiaj lim dej, lub laser (xiav) yog tsav hla lub fiber ntau optic cable los ntawm FC / APC connector.Lub beam yog deflected los ntawm ib daim iav dichroic (DM) rau hauv ib hom fiber ntau (SMF) ntawm tus thib ob FC / APC connector.Fluorescence (ntsuab) taug kev los ntawm DM thiab ntev dhau lim (LPF) mus rau PMT ntawm multimode fiber (MMF).(c) Qhov kawg ntawm qhov endoscope txuas nrog fiber ntau optic (F) thiab wired (W) ports ntawm lub hauv paus chaw nres tsheb.(d) Endoscope, saib, chaw nres tsheb hauv paus, khoos phis tawj, thiab rho tawm hloov pauv ntawm lub tsheb thauj khoom.(a, c) Solidworks 2016 tau siv rau 3D qauv ntawm cov duab kos duab thiab cov khoom siv endoscope.
Lub lateral thiab axial daws teeb meem ntawm lub tsom optics tau ntsuas los ntawm qhov taw tes kis kev ua haujlwm ntawm fluorescent microspheres (#F8803, Thermo Fisher Scientific) 0.1 µm inch.Sau cov duab los ntawm kev txhais cov microspheres horizontally thiab vertically hauv 1 µm cov kauj ruam siv ib theem kab (# M-562-XYZ, DM-13, Newport).Cov duab pawg siv ImageJ2 kom tau txais cov duab sib luag ntawm microspheres.
Cov software tshwj xeeb (LabVIEW 2021, NI) tau tsim los rau kev nrhiav cov ntaub ntawv ntawm lub sijhawm thiab ua cov duab.Ntawm daim duab.7 qhia txog lub ntsiab lus ntawm cov txheej txheem uas siv los khiav qhov system.Tus neeg siv interface muaj cov ntaub ntawv tau txais (DAQ), lub ntsiab vaj huam sib luag thiab cov tswj vaj huam sib luag.Lub vaj huam sib luag sau cov ntaub ntawv cuam tshuam nrog lub ntsiab vaj huam sib luag los sau thiab khaws cov ntaub ntawv nyoos, muab cov tswv yim rau kev cai sau cov ntaub ntawv, thiab tswj cov chaw tsav tsheb scanner.Lub vaj huam sib luag tseem ceeb tso cai rau tus neeg siv xaiv qhov xav tau kev teeb tsa rau kev siv lub tshuab endoscope, suav nrog lub tshuab luam ntawv tswj lub teeb liab, video ncej tus nqi, thiab cov khoom tsis tau.Lub vaj huam sib luag no tseem tso cai rau tus neeg siv los tso saib thiab tswj qhov ci ntsa iab thiab qhov sib txawv ntawm cov duab.Siv cov ntaub ntawv nyoos raws li cov tswv yim, lub algorithm xam qhov pom kev nce qhov chaw rau PMT thiab hloov kho qhov ntsuas no siv qhov proportional-integral (PI)16 tawm tswv yim tswj qhov system.Lub rooj tsav xwm tswj tau cuam tshuam nrog lub rooj tsav xwm tseem ceeb thiab cov ntaub ntawv nrhiav tau los tswj lub zog laser thiab PMT nce.
System software architecture.Tus neeg siv interface muaj cov modules (1) cov ntaub ntawv nrhiav tau (DAQ), (2) lub ntsiab vaj huam sib luag thiab (3) tswj vaj huam sib luag.Cov kev pab cuam no ua haujlwm ib txhij thiab sib txuas lus nrog ib leeg los ntawm cov kab lus.Qhov tseem ceeb yog MEMS: Microelectromechanical System, TDMS: Technical Data Control Flow, PI: Proportional Integral, PMT: Photomultiplier.Cov duab thiab video ntaub ntawv tau txais kev cawmdim nyob rau hauv BMP thiab AVI hom, feem.
Ib theem kho algorithm yog siv los xam cov dispersion ntawm cov duab pixel siv ntawm cov theem sib txawv los txiav txim siab tus nqi siab tshaj plaws uas siv los ua kom cov duab.Rau kev kho lub sijhawm tiag tiag, theem scan ntau yog ± 2.86 ° nrog cov kauj ruam loj ntawm 0.286 ° kom txo cov sijhawm suav.Tsis tas li ntawd, siv qhov chaw ntawm daim duab nrog cov qauv tsawg dua ntxiv txo cov duab ncej suav lub sij hawm los ntawm 7.5 vib nas this (1 Msample) mus rau 1.88 vib nas this (250 Ksample) ntawm 10 Hz.Cov input parameters no tau raug xaiv los muab cov duab zoo txaus nrog tsawg latency thaum nyob hauv vivo imaging.Cov duab nyob thiab cov yeeb yaj kiab raug kaw hauv BMP thiab AVI hom, feem.Cov ntaub ntawv nyoos yog khaws cia rau hauv Technical Data Management Flow Format (TMDS).
Tom qab ua tiav ntawm cov duab hauv vivo rau kev txhim kho kom zoo nrog LabVIEW 2021. Qhov tseeb raug txwv thaum siv cov txheej txheem kho cov theem thaum nyob hauv vivo imaging vim lub sijhawm suav ntev xav tau.Tsuas yog cov duab txwv tsis pub siv thiab cov lej piv txwv xwb.Tsis tas li ntawd, lub algorithm tsis ua haujlwm zoo rau cov duab nrog cov duab kos duab los yog qhov sib txawv tsis zoo thiab ua rau cov theem kev suav ua yuam kev30.Cov thav duab ntawm tus kheej nrog qhov sib txawv siab thiab tsis muaj cov khoom siv txav tau raug xaiv los ntawm cov theem zoo tuning nrog rau theem scan ntau ntawm ± 0.75 ° hauv 0.01 ° cov kauj ruam.Tag nrho cov duab cheeb tsam tau siv (piv txwv li, 1 Msample ntawm ib daim duab kaw ntawm 10 Hz).Table S2 qhia txog cov duab tsis siv rau lub sijhawm tiag tiag thiab kev ua haujlwm tom qab.Tom qab kho theem, lub lim nruab nrab yog siv los txo cov duab nrov ntxiv.Brightness thiab sib piv yog ntxiv txhim kho los ntawm histogram stretching thiab gamma kho31.
Cov kev sim tshuaj tau pom zoo los ntawm Michigan Medical Institutions Review Board thiab tau ua nyob rau hauv Lub Tsev Haujlwm Saib Xyuas Kev Kho Mob.Txoj kev tshawb no tau sau npe online nrog ClinicalTrials.gov (NCT03220711, hnub sau npe: 07/18/2017).Cov txheej txheem suav nrog suav nrog cov neeg mob (hnub nyoog 18 txog 100 xyoo) nrog kev npaj ua ntej xaiv txoj hnyuv loj, muaj kev pheej hmoo ntawm mob qog noj ntshav, thiab keeb kwm ntawm kab mob plab hnyuv.Cov ntaub ntawv tso cai tau txais los ntawm txhua qhov kev kawm uas pom zoo koom nrog.Cov txheej txheem cais tawm yog cov neeg mob uas cev xeeb tub, muaj kev paub tsis meej rau fluorescein, lossis tau txais kev siv tshuaj khomob lossis kev kho hluav taws xob.Txoj kev tshawb fawb no suav nrog cov neeg mob sib law liag tau teem sijhawm rau kev kuaj mob colonoscopy thiab yog tus sawv cev ntawm Michigan Medical Center cov pej xeem.Txoj kev tshawb no tau ua raws li Kev Tshaj Tawm ntawm Helsinki.
Ua ntej kev phais, calibrate lub endoscope siv 10 µm fluorescent hlaws dai (#F8836, Thermo Fisher Scientific) mounted nyob rau hauv silicone pwm.Lub translucent silicone sealant (#RTV108, Momentive) tau nchuav rau hauv 3D luam tawm 8 cm3 yas pwm.Tso cov hlaws dej fluorescent hla lub silicone thiab tawm mus txog thaum dej nruab nrab dries.
Tag nrho cov nyuv raug tshuaj xyuas siv tus qauv kho mob colonoscope (Olympus, CF-HQ190L) nrog lub teeb dawb.Tom qab tus kws kho mob endoscopist tau txiav txim siab thaj tsam ntawm tus kab mob, thaj chaw yog ntxuav nrog 5-10 ml ntawm 5% acetic acid, thiab tom qab ntawd nrog cov dej tsis huv kom tshem tawm cov hnoos qeev thiab cov khib nyiab.Ib koob tshuaj 5 ml ntawm 5 mg / ml fluorescein (Alcon, Fluorescite) tau txhaj tshuaj intravenously los yog tsuag tshuaj pleev xim rau hauv cov mucosa siv tus qauv cannula (M00530860, Boston Scientific) uas tau dhau los ntawm txoj haujlwm ua haujlwm.
Siv lub irrigator los yaug cov xim ntau dhau los yog cov khib nyiab los ntawm qhov chaw mucosal.Tshem lub nebulizing catheter thiab dhau qhov endoscope los ntawm kev ua haujlwm channel kom tau txais cov duab ante-mortem.Siv cov kev taw qhia dav dav endoscopic los muab cov lus qhia ntawm qhov chaw nyob rau hauv lub hom phiaj. Lub sijhawm tag nrho siv los sau cov duab confocal yog <10 min. Lub sijhawm tag nrho siv los sau cov duab confocal yog <10 min. Общее время, затраченное на сбор конфокальных изображений, составило <10 мин. Lub sijhawm tag nrho coj los sau cov duab confocal yog <10 min.Lub sijhawm tau txais tag nrho rau cov duab confocal tsawg dua 10 feeb.Endoscopic dawb lub teeb video tau ua tiav siv Olympus EVIS EXERA III (CLV-190) cov duab thiab kaw siv Elgato HD video recorder.Siv LabVIEW 2021 los sau thiab khaws cov yeeb yaj kiab endoscopy.Tom qab cov duab tiav lawm, qhov endoscope raug tshem tawm thiab cov ntaub so ntswg kom pom yog excised siv biopsy forceps los yog ib tug snare. Cov ntaub so ntswg tau ua tiav rau niaj hnub histology (H&E), thiab soj ntsuam los ntawm tus kws tshaj lij GI pathologist (HDA). Cov ntaub so ntswg tau ua tiav rau niaj hnub histology (H&E), thiab soj ntsuam los ntawm tus kws tshaj lij GI pathologist (HDA). Ткани были обработаны для обычной гистологии (H&E) и оценены экспертом-патологом желудочно-кишечногко Cov ntaub so ntswg tau ua tiav rau niaj hnub histology (H&E) thiab soj ntsuam los ntawm tus kws kho mob plab hnyuv siab raum (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) 进行评估.对组织进行常规组织学(H&E) 处理，并由专家GI 病理学家(HDA) 进行评估. Ткани были обработаны для обычной гистологии (H&E) и оценены экспертом-патологом желудочно-кишечногко Cov ntaub so ntswg tau ua tiav rau niaj hnub histology (H&E) thiab soj ntsuam los ntawm tus kws kho mob plab hnyuv siab raum (HDA).Cov khoom spectral ntawm fluorescein tau lees paub siv lub spectrometer (USB2000+, Ocean Optics) raws li qhia hauv daim duab S2.
Endoscopes tau ua kom tsis muaj menyuam tom qab txhua tus neeg siv (Fig. 8).Cov txheej txheem ntxuav tau ua raws li kev coj ua thiab kev pom zoo ntawm Lub Tsev Haujlwm Saib Xyuas Kev Tiv Thaiv Kab Mob thiab Kab Mob ntawm Michigan Medical Center thiab Central Sterile Processing Unit. Ua ntej txoj kev tshawb fawb, cov cuab yeej tau sim thiab siv tau rau kev ua kom tsis muaj menyuam los ntawm Advanced Sterilization Products (ASP, Johnson & Johnson), lub chaw lag luam uas muab kev tiv thaiv kab mob thiab kev ua kom tsis muaj menyuam. Ua ntej txoj kev tshawb fawb, cov cuab yeej tau sim thiab siv tau rau kev ua kom tsis muaj menyuam los ntawm Advanced Sterilization Products (ASP, Johnson & Johnson), lub chaw lag luam uas muab kev tiv thaiv kab mob thiab kev ua kom tsis muaj menyuam. Перед исследованием инструменты были протестированы и одобрены для стерилизации компанией Advanced Sterilization Products, Johnson (ASP, Johnson) ией, предоставляющей услуги по профилактике инфекций и проверке стерилизации. Ua ntej kev kawm, cov cuab yeej raug kuaj thiab pom zoo rau kev ua kom tsis muaj menyuam los ntawm Advanced Sterilization Products (ASP, Johnson & Johnson), lub koom haum kev lag luam uas muab kev pabcuam tiv thaiv kab mob thiab ua kom tsis muaj menyuam. Перед исследованием инструменты были стерилизованы и проверены Advanced Sterilization Products (ASP, Johnson & Johnson), коммерческой орграния цатрания услуги по профилактике инфекций и проверке стерилизации. Cov cuab yeej tau ua kom tsis muaj menyuam thiab tshuaj xyuas ua ntej kev kawm los ntawm Advanced Sterilization Products (ASP, Johnson & Johnson), lub koom haum ua lag luam uas muab kev pabcuam tiv thaiv kab mob thiab ua kom tsis muaj menyuam.
Cov cuab yeej siv dua tshiab.(a) Endoscopes tau muab tso rau hauv cov tais tom qab txhua qhov ua kom tsis muaj menyuam siv cov txheej txheem STERRAD.(b) SMF thiab cov xov hlau raug txiav nrog fiber ntau optic thiab hluav taws xob connectors, raws li, uas raug kaw ua ntej yuav rov ua dua.
Ntxuav lub endoscopes los ntawm kev ua cov hauv qab no: (1) so lub endoscope nrog ib tug lint-dawb daim ntaub soaked nyob rau hauv ib tug enzymatic nqus tsev vacuum los ntawm proximal mus rau distal;(2) Tso cov twj tso rau hauv cov tshuaj detergent enzymatic rau 3 feeb nrog dej.ntaub tsis muaj lint.Cov khoom siv hluav taws xob thiab fiber ntau optic tau them thiab tshem tawm ntawm kev daws teeb meem;(3) Lub endoscope yog qhwv thiab muab tso rau hauv lub tais ntsuas kev ua kom tsis muaj menyuam siv STERRAD 100NX, hydrogen peroxide roj ntshav plasma.kuj tsis tshua muaj kub thiab tsawg humidity ib puag ncig.
Cov ntaub ntawv siv thiab / lossis tshuaj ntsuam hauv txoj kev tshawb fawb tam sim no muaj los ntawm cov kws sau ntawv raws li qhov kev thov tsim nyog.
Pilonis, ND, Januszewicz, W. & di Pietro, M. Confocal laser endomicroscopy hauv gastro-testinal endoscopy: Cov yam ntxwv thiab kev siv tshuaj kho mob. Pilonis, ND, Januszewicz, W. & di Pietro, M. Confocal laser endomicroscopy hauv gastro-testinal endoscopy: Cov yam ntxwv thiab kev siv tshuaj kho mob.Pilonis, ND, Januszewicz, V. i di Pietro, M. Confocal laser endomicroscopy in gastrointestinal endoscopy: technicalities and clinical application. Pilonis, ND, Januszewicz, W. & di Pietro, M. 胃肠 的更多内容 Pilonis, ND, Januszewicz, W. & di Pietro, M.Pilonis, ND, Januszewicz, V. i di Pietro, M. Confocal laser endoscopy in gastrointestinal endoscopy: technicalities and clinical applications.txhais lus gastrointestinal heparin.7, 7 (2022).
Al-Mansour, MR et al.Kev nyab xeeb thiab kev ua tau zoo ntawm SAGES TAVAC Confocal Laser Endomicroscopy.Kev ua haujlwm.Endoscopy 35, 2091–2103 (2021).
Fugazza, A. et al.Confocal laser endoscopy nyob rau hauv plab hnyuv thiab pancreatobiliary kab mob: ib tug systematic tshuaj xyuas thiab meta-analysis.Biomedical Science.cia tank.sab hauv 2016, 4638683 (2016).