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Confocal laser endoscopy ndi njira yatsopano yowonera zenizeni zenizeni zenizeni.Zithunzi za fluorescent za khalidwe la histological zitha kupezeka nthawi yomweyo kuchokera ku epithelium ya ziwalo zopanda kanthu.Pakadali pano, kusanthula kumachitika motsatana ndi zida zozikidwa pa probe zomwe zimagwiritsidwa ntchito kwambiri pazachipatala, komanso kusinthasintha kochepa pakuwongolera kuyang'ana.Tikuwonetsa kugwiritsa ntchito makina ojambulira a parametric omwe amayikidwa kumapeto kwa endoscope kuti atembenuke mothamanga kwambiri.Bowo lazikika pakati pa chounikira kuti likulungitse njira yowunikira.Kapangidwe kameneka kamachepetsa kukula kwa chidacho kukhala 2.4 mm m'mimba mwake ndi 10 mm m'litali, kulola kupititsidwa patsogolo kudzera munjira yogwirira ntchito ya ma endoscopes achipatala.Ma lens ophatikizika amapereka kusinthasintha kwapakati ndi axial kwa 1.1 ndi 13.6 µm, motsatana.Mtunda wogwira ntchito wa 0 µm ndi malo owonera 250 µm × 250 µm amakwaniritsidwa pamitengo mpaka 20 Hz.Kusangalatsa kwa 488 nm kumasangalatsa fluorescein, utoto wovomerezeka ndi FDA wosiyanitsa kwambiri minofu.Ma endoscopes adasinthidwanso kwa mizere 18 popanda kulephera pogwiritsa ntchito njira zovomerezeka zachipatala.Zithunzi za fluorescent zinapezedwa kuchokera ku colonic mucosa, tubular adenomas, hyperplastic polyps, ulcerative colitis, ndi Crohn's colitis panthawi ya colonoscopy.Maselo amodzi amatha kudziwika, kuphatikiza ma colonocyte, ma cell a goblet, ndi maselo otupa.Zinthu za mucosal monga mapangidwe a crypt, crypt cavities, ndi lamina propria zitha kuzindikirika.Chidacho chingagwiritsidwe ntchito ngati chothandizira ku endoscopy yanthawi zonse.
Confocal laser endoscopy ndi njira yatsopano yojambula yomwe ikupangidwira kuti igwiritsidwe ntchito pachipatala monga chothandizira ku endoscopy1,2,3.Zida zosinthika, zolumikizidwa ndi fiber-optic zitha kugwiritsidwa ntchito pozindikira matenda omwe ali m'maselo a epithelial omwe amatsata ziwalo zopanda kanthu, monga m'matumbo.Minofu yopyapyalayi imakhala yogwira ntchito kwambiri ndipo ndi yomwe imayambitsa matenda ambiri monga khansa, matenda, ndi kutupa.Endoscopy imatha kukwaniritsa kusamvana kwapang'onopang'ono, kupereka zenizeni zenizeni, pafupifupi za mbiri yakale muzithunzi za vivo kuthandiza asing'anga kupanga zosankha zachipatala.Biopsy ya minofu yamthupi imakhala ndi chiopsezo chotaya magazi komanso kuphulika.Zitsanzo zambiri kapena zochepa za biopsy nthawi zambiri zimasonkhanitsidwa.Chitsanzo chilichonse chochotsedwa chimawonjezera mtengo wa opaleshoni.Zimatenga masiku angapo kuti chitsanzocho chiwunikidwe ndi dokotala.M'masiku oyembekezera zotsatira za pathology, odwala nthawi zambiri amakhala ndi nkhawa.Mosiyana ndi zimenezi, njira zina zowonongeka zachipatala monga MRI, CT, PET, SPECT, ndi ultrasound zimasowa kusintha kwa malo ndi liwiro lakanthawi lofunika kuti liwonetsetse njira za epithelial mu vivo ndi nthawi yeniyeni, kusamvana kwa subcellular.
Chipangizo chopangidwa ndi probe (Cellvizio) chimagwiritsidwa ntchito kwambiri m'zipatala popanga "optical biopsy".Kapangidwe kake kamachokera pamtundu wa fiber optic bundle4 womwe umasonkhanitsa ndikutumiza zithunzi za fulorosenti.Chingwe cha fiber core chimagwira ntchito ngati "dzenje" kuti zisefa kuwala kosasunthika kuti zisawonongeke.Kusanthula kumachitika pafupi ndi galvanometer yayikulu, yokulirapo.Kupereka uku kumachepetsa kuthekera kwa chida chowongolera.Kuyika koyenera kwa epithelial carcinoma kumafuna kuyang'ana pansi pa minofu kuti awone kuukira ndikupeza chithandizo choyenera.Fluorescein, wothandizira wovomerezeka ndi FDA, amaperekedwa kudzera m'mitsempha kuti awonetse mawonekedwe a epithelium. Ma endomicroscopes amenewa ali ndi miyeso <2.4 mm m'mimba mwake, ndipo amatha kupitilira patsogolo mosavuta kudzera mu njira ya biopsy ya ma endoscopes achipatala. Ma endomicroscopes amenewa ali ndi miyeso <2.4 mm m'mimba mwake, ndipo amatha kupitilira patsogolo mosavuta kudzera mu njira ya biopsy ya ma endoscopes achipatala. Эти эндомикроскопы имеют размеры <2,4 мм в диаметре ndi могут быть легко проведены через биопсийный канал стандартных медицински медицински. Ma endomicroscopes amenewa ndi <2.4 mm m'mimba mwake ndipo amatha kudutsa mosavuta kudzera mu njira ya biopsy ya ma endoscopes achipatala.Ma borescopes amenewa ndi osakwana mamilimita 2.4 m'mimba mwake ndipo amadutsa mosavuta panjira ya biopsy ya borescopes wamba.Kusinthasintha kumeneku kumapangitsa kuti pakhale ntchito zambiri zachipatala ndipo ndizodziyimira pawokha kwa opanga ma endoscope.Kafukufuku wambiri wazachipatala achitidwa pogwiritsa ntchito chipangizo chojambulachi, kuphatikiza kuzindikira koyambirira kwa khansa yam'mero, m'mimba, m'matumbo, ndi pakamwa.Ma protocol amajambula apangidwa ndipo chitetezo cha njirayi chakhazikitsidwa.
Microelectromechanical systems (MEMS) ndi ukadaulo wamphamvu popanga ndi kupanga tinthu tating'onoting'ono tosanthula zomwe zimagwiritsidwa ntchito kumapeto kwa ma endoscopes.Udindo uwu (wokhudzana ndi proximal) umalola kusinthasintha kwakukulu pakuwongolera malo owonetsetsa5,6.Kuphatikiza pa kupotoza kwapambuyo, makina a distal amathanso kupanga ma axial scans, post-objective scans, ndi ma scans opezeka mwachisawawa.Kuthekera kumeneku kumathandizira kufufuzidwa mozama kwa ma cell a epithelial, kuphatikiza chithunzithunzi chapakatikati7, mawonekedwe akulu (FOV)8 osapanga sikani, komanso magwiridwe antchito abwino m'magawo ang'onoang'ono9.MEMS imathetsa vuto lalikulu lakuyika injini yojambulira ndi malo ochepa omwe amapezeka kumapeto kwa chidacho.Poyerekeza ndi ma galvanometer ochulukirapo, MEMS imapereka magwiridwe antchito apamwamba pang'ono, kuthamanga kwambiri, komanso kugwiritsa ntchito mphamvu zochepa.Njira yosavuta yopangira ikhoza kuwonjezeredwa kuti ikhale yochuluka pamtengo wotsika.Mapangidwe ambiri a MEMS adanenedwa kale10,11,12.Palibe matekinoloje omwe adapangidwa mokwanira kuti athe kufalikira kwachipatala kwa nthawi yeniyeni mu kujambula kwa vivo kudzera munjira yogwirira ntchito ya endoscope yachipatala.Pano, tikufuna kuwonetsa kugwiritsidwa ntchito kwa scanner ya MEMS kumapeto kwa endoscope kuti apeze zithunzi za anthu mu vivo panthawi yachipatala endoscope.
Chida cha fiber optic chinapangidwa pogwiritsa ntchito scanner ya MEMS kumapeto kwakutali kuti atenge zithunzi zenizeni za vivo fulorosenti zokhala ndi mawonekedwe ofanana a histological.Chingwe cha single-mode (SMF) chimatsekeredwa mu chubu chosinthika cha polima ndikusangalatsidwa pa λex = 488 nm.Kukonzekera uku kumafupikitsa kutalika kwa nsonga yakutali ndikulola kuti idutsidwe kudzera munjira yogwirira ntchito ya ma endoscopes achipatala.Gwiritsani nsongayo kuti mukhazikike pakati pa optic.Magalasi awa adapangidwa kuti akwaniritse pafupifupi diffractive axial resolution yokhala ndi kabowo kakang'ono (NA) = 0.41 ndi mtunda wogwirira ntchito = 0 µm13.Ma shims olondola amapangidwa kuti agwirizane bwino ndi optics 14. Chojambuliracho chimayikidwa mu endoscope ndi nsonga yolimba ya distal 2.4 mm m'mimba mwake ndi 10 mm kutalika (mkuyu 1a).Miyeso iyi imalola kuti igwiritsidwe ntchito muzochita zachipatala monga chothandizira pa endoscopy (mkuyu 1b).Mphamvu yayikulu ya chochitika cha laser pa minofu inali 2 mW.
Confocal laser endoscopy (CLE) ndi makina a MEMS.Chithunzi chosonyeza (a) chida chopakidwa chomwe chili ndi nsonga yolimba ya nsonga ya mamilimita 2.4 ndi utali wa mamilimita 10 ndi (b) njira yowongoka yodutsa munjira yogwirira ntchito ya endoscope yamankhwala yokhazikika (Olympus CF-HQ190L).(c) Kuyang'ana kutsogolo kwa scanner yowonetsa chonyezimira chokhala ndi pobowo pakati pa 50 µm momwe mtengo wokokera umadutsa.Chojambuliracho chimayikidwa pa gimbal yoyendetsedwa ndi ma drive a quadrature comb drive.Kuchuluka kwa resonant kwa chipangizocho kumatsimikiziridwa ndi kukula kwa kasupe wa torsion.(d) Mawonedwe am'mbali a scanner yowonetsa sikelo yoyikidwa pa choyimira chokhala ndi mawaya olumikizidwa ndi nangula wa ma elekitirodi omwe amapereka malo olumikizirana ndi ma drive ndi ma siginecha amagetsi.
Makina ojambulira amakhala ndi chowunikira chopangidwa ndi gimbal chomwe chimayendetsedwa ndi ma quadrature actuators oyendetsedwa ndi chisa kuti asokoneze mtengowo (ndege ya XY) munjira ya Lissajous (mkuyu 1c).Bowo la 50 µm m'mimba mwake lidakhazikika pakati pomwe mtengo wokokerawo udadutsa.Chojambuliracho chimayendetsedwa ndi ma frequency a resonant a mapangidwe, omwe amatha kusinthidwa posintha miyeso ya kasupe wa torsion.Nangula wa Electrode adalembedwa pamphepete mwa chipangizocho kuti apereke mfundo zolumikizirana ndi mphamvu ndi zidziwitso zowongolera (mkuyu 1d).
Makina ojambulira amayikidwa pangolo yonyamula yomwe imatha kukulungidwa m'chipinda chopangira opaleshoni.Mawonekedwe azithunzi adapangidwa kuti azithandizira ogwiritsa ntchito omwe ali ndi chidziwitso chochepa chaukadaulo, monga madotolo ndi anamwino.Yang'anani pamanja ma frequency a scanner drive, beamform mode, ndi chithunzi cha FOV.
Utali wonse wa endoscope ndi pafupifupi 4m kulola zida zonse kudutsa munjira yogwirira ntchito ya endoscope yachipatala yokhazikika (1.68m), yokhala ndi kutalika kowonjezera pakuwongolera.Pamapeto oyandikira endoscope, ma SMF ndi mawaya amatha mu zolumikizira zomwe zimalumikizana ndi madoko a fiber optic ndi mawaya a siteshoni yoyambira.Kuyikako kumakhala ndi laser, unit fyuluta, amplifier high-voltage ndi photomultiplier detector (PMT).Amplifier imapereka mphamvu ndikuyendetsa ma siginecha ku scanner.Chigawo cha kuwala chimagwirizanitsa chisangalalo cha laser ku SMF ndikudutsa fluorescence ku PMT.
Ma endoscopes amakonzedwanso pambuyo pa njira iliyonse yachipatala pogwiritsa ntchito njira yotseketsa ya STERRAD ndipo amatha kupirira mpaka 18 popanda kulephera.Pa yankho la OPA, palibe zisonyezo za kuwonongeka zomwe zidawonedwa pambuyo pazaka zopitilira 10 zopha tizilombo.Zotsatira za OPA zidapambana ma STERRAD, kutanthauza kuti moyo wa ma endoscopes utha kuwonjezedwa ndi kupha tizilombo toyambitsa matenda m'malo moletsanso kubereka.
Kusintha kwazithunzi kudatsimikiziridwa kuchokera pakufalikira kwa mfundo pogwiritsa ntchito mikanda ya fulorosenti yokhala ndi mainchesi a 0.1 μm.Kwa lateral ndi axial kusamvana, m'lifupi lonse pa theka lapamwamba (FWHM) la 1.1 ndi 13.6 µm, motero, linayesedwa (mkuyu 2a, b).
Zithunzi zosankha.The lateral (a) ndi axial (b) kusamvana kwa kuyang'ana Optics amadziwika ndi mfundo kufalikira ntchito (PSF) kuyezedwa pogwiritsa ntchito fulorosenti microspheres ndi awiri a 0.1 μm.Kuyeza kwathunthu m'lifupi pa theka lapamwamba (FWHM) kunali 1.1 ndi 13.6 µm, motsatira.Choyikapo: Mawonedwe owonjezera a microsphere imodzi modutsa (XY) ndi axial (XZ) akuwonetsedwa.(c) Chithunzi cha fluorescent chopezedwa kuchokera ku mzere (USAF 1951) chandamale (chozungulira chofiira) chosonyeza kuti magulu 7-6 akhoza kuthetsedwa bwino.(d) Chithunzi cha 10 µm m'mimba mwake mwa ma microspheres obalalitsidwa a fulorosenti kusonyeza malo azithunzi a 250 µm×250 µm.Ma PSF mu (a, b) adamangidwa pogwiritsa ntchito MATLAB R2019a (https://www.mathworks.com/).(c, d) Zithunzi za fluorescent zidasonkhanitsidwa pogwiritsa ntchito LabVIEW 2021 (https://www.ni.com/).
Fluorescent zithunzi kuchokera muyezo kusamvana magalasi bwino kusiyanitsa ya mizati mu magulu 7-6, amene amasunga mkulu lateral kusamvana (mkuyu. 2c).Malo owonera (FOV) a 250 µm × 250 µm adatsimikiziridwa kuchokera ku zithunzi za mikanda ya fulorosenti ya 10 µm m'mimba mwake yomwazikana pa zophimba (mkuyu 2d).
Njira yodzichitira yokha ya PMT kuwongolera ndikuwongolera gawo kumayendetsedwa mukamaganizira zachipatala kuti muchepetse zinthu zoyenda kuchokera ku endoscopes, colon peristalsis, ndi kupuma kwa odwala.Kumanganso zithunzi ndi kukonza ma algorithms afotokozedwa kale14,15.Kupindula kwa PMT kumayendetsedwa ndi wolamulira wa proportional-integral (PI) kuti apewe kuchuluka kwamphamvu16.Dongosolo limawerenga kuchuluka kwa pixel kwa chimango chilichonse, kuwerengera mayankho ofananirako komanso ofunikira, ndikusankha mapindu a PMT kuti awonetsetse kuti kukula kwa pixel kuli mkati mwazovomerezeka.
Panthawi yoyerekeza mu vivo, kusagwirizana kwa gawo pakati pa kayendedwe ka sikani ndi chizindikiro chowongolera kungayambitse kusawoneka bwino.Zotsatira zoterezi zingachitike chifukwa cha kusintha kwa kutentha kwa chipangizo mkati mwa thupi la munthu.Zithunzi zoyera zoyera zidawonetsa kuti endoscope idalumikizana ndi mucosa wamba mu vivo (Chithunzi 3a).Kusawoneka bwino kwa ma pixel osokonekera kumatha kuwoneka pazithunzi zosaphika za mucosa wamba (Chithunzi 3b).Pambuyo pa chithandizo ndi gawo loyenera komanso kusintha kosiyana, mawonekedwe a subcellular a mucosa amatha kusiyanitsa (mkuyu 3c).Kuti mudziwe zambiri, zithunzi za confocal yaiwisi ndi zithunzi zowonongeka zenizeni zikuwonetsedwa mu Mkuyu.
Kukonza zithunzi.(a) Chithunzi cha endoscope chotalikirapo chomwe chikuwonetsa endoscope (E) yolumikizidwa ndi mucosa wamba (N) colonic mucosa kuti itenge zithunzi za fulorosenti mu vivo pambuyo poyendetsa fluorescein.(b) Kuyendayenda mu ma ax a X ndi Y panthawi ya scanning kungapangitse ma pixel osokonekera kuti asokonezeke.Zolinga zowonetsera, kusintha kwakukulu kumagwiritsidwa ntchito pa chithunzi choyambirira.(c) Pambuyo pokonza gawo lokonzekera, tsatanetsatane wa mucosal akhoza kuyesedwa, kuphatikizapo mapangidwe a crypt (mivi), ndi lumen yapakati (l) yozunguliridwa ndi lamina propria (lp).Maselo amodzi amatha kusiyanitsa, kuphatikiza ma colonocyte (c), ma cell a goblet (g), ndi maselo otupa (mivi).Onani kanema wowonjezera 1. (b, c) Zithunzi zokonzedwa pogwiritsa ntchito LabVIEW 2021.
Zithunzi za confocal fluorescence zapezeka mu vivo m'matenda angapo a colonic kuwonetsa kugwiritsa ntchito kwachipatala kwa chidacho.Kujambula kwa mbali zotalikirana kumachitidwa koyamba pogwiritsa ntchito kuwala koyera kuti azindikire ntchofu zachilendo kwambiri.Kenako endoscope imatsogozedwa ndi njira yogwirira ntchito ya colonoscope ndikulumikizana ndi mucosa.
Wide-field endoscopy, confocal endomicroscopy, ndi histology (H&E) zithunzi zimawonetsedwa za colonic neoplasia, kuphatikiza tubular adenoma ndi hyperplastic polyp. Wide-field endoscopy, confocal endomicroscopy, ndi histology (H&E) zithunzi zimawonetsedwa za colonic neoplasia, kuphatikiza tubular adenoma ndi hyperplastic polyp. Широкопольная эндоскопия, конфокальная эндомикроскопия ndi гистологические (H&E) ndi зображения показаны для неоплазии токублязия токирустоя в песни у ndi гиперпластический полип. Colonic endoscopy, confocal endomicroscopy, ndi histological (H&E) imaging amasonyezedwa pa colonic neoplasia, kuphatikiza tubular adenoma ndi 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 管状 管状 管状 躰化 增生性息肉 增生性息肉 增生性息肉 增生性息肉 的 内刵霱录 内刵霱录 共 共 在 在 微微全 在 具 具 具 具 具 具 (H & E Широкопольная эндоскопия, конфокальная микроэндоскопия ndi гистологические (H&E) ndi зображения, конфокальная микроэндоскопия ndi гистологические (H&E) ndi зображения, конфокальная микроэндоскопия ndi гистологические (H&E) ndi зображения, показывающие опухоскопия опухолика опухолики той, конфокальная микроэндоскопия ndi гистологические (H&E) мы ndi гиперпластические полипы. Broad-field endoscopy, confocal microendoscopy, ndi histological (H&E) zithunzi zowonetsa zotupa zam'matumbo, kuphatikiza ma tubular adenomas ndi hyperplastic polyps.Ma tubular adenomas adawonetsa kutayika kwa kamangidwe kabwino ka crypt, kuchepa kwa kukula kwa ma cell a goblet, kupotoza kwa lumen ya crypt, ndikukula kwa lamina propria (mkuyu 4a-c).Ma polyps a hyperplastic adawonetsa mapangidwe a stellate a ma crypts, maselo ochepa a goblet, lumen-ngati lumen of crypts, ndi ma lamellar crypts osakhazikika (mkuyu 4d-f).
Chithunzi cha mucosal thick skin in vivo. Oimira kuwala koyera endoscopy, confocal endomicroscope, ndi histology (H&E) zithunzi za (ac) adenoma, (df) hyperplastic polyp, (gi) ulcerative colitis, ndi (jl) Crohn's colitis. Oimira kuwala koyera endoscopy, confocal endomicroscope, ndi histology (H&E) zithunzi za (ac) adenoma, (df) hyperplastic polyp, (gi) ulcerative colitis, ndi (jl) Crohn's colitis. Типичные изображения эндоскопии в белом свете, конфокального эндомикроскопа ndi гистологии (H&E) показаны для (ac) аденомы, показаны (ac) аденомы, показаны (df). нного колита и (jl) колита Крона. Zithunzi zowoneka bwino zokhala ndi kuwala koyera, confocal endomicroscope, ndi histology (H&E) zithunzi za (ac) adenoma, (df) hyperplastic polyp, (gi) ulcerative colitis, ndi (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) 克罗恩结肠炎的代腨全性聚焦内窥镜检查和组织学( H&E) 图像. Zimasonyeza(ac) 躰真、(df) 增生性息肉、(gi) 苏盖性红肠炎和(jl) 克罗恩红肠炎的体育性白光、性白光内肠肠炎性和电视学( H&E ) chithunzi. Представлены репрезентативные эндоскопия в белом свете, конфокальная эндоскопия (ac) аденомы, (df) гипезарплика, конфокальная эндоскопия (ac) аденомы, (df) гипезаркоплика l) колита Крона (H&E). Oimira white-light endoscopy, confocal endoscopy, ndi histology ya (ac) adenoma, (df) hyperplastic polyposis, (gi) ulcerative colitis, ndi (jl) Crohn's colitis (H&E) akuwonetsedwa.(B) akuwonetsa chithunzi cholumikizira chopezeka mu vivo kuchokera ku tubular adenoma (TA) pogwiritsa ntchito endoscope (E).Chotupa chowopsachi chikuwonetsa kutayika kwa kamangidwe kabwinobwino ka crypt (muvi), kupotoza kwa crypt lumen (l), komanso kuchuluka kwa crypt lamina propria (lp).Ma Colonocytes (c), ma cell a goblet (g), ndi maselo otupa (mivi) amathanso kudziwika.Smt.Kanema Wowonjezera 2. (e) akuwonetsa chithunzi cha confocal chopezedwa kuchokera ku hyperplastic polyp (HP) mu vivo.Chotupa ichi chikuwonetsa kamangidwe ka stellate crypt (muvi), lumen ngati crypt (l), ndi lamina propria (lp).Ma Colonocyte (c), ma cell a goblet angapo (g) ndi maselo otupa (mivi) amathanso kudziwika.Smt.Kanema Wowonjezera 3. (h) akuwonetsa zithunzi za confocal zomwe zimapezeka mu ulcerative colitis (UC) mu vivo.Kutupa kumeneku kumawonetsa kamangidwe kolakwika ka crypt (muvi) ndi ma cell odziwika bwino a goblet (g).Nthenga za fluorescein (f) zimatuluka m'maselo a epithelial, kuwonetsa kuchuluka kwa mitsempha yamagazi.Maselo ambiri otupa (mivi) amawoneka mu lamina propria (lp).Smt.Kanema Wowonjezera 4. (k) akuwonetsa chithunzi cholumikizira chopezeka mu vivo kuchokera kudera la Crohn's colitis (CC).Kutupa kumeneku kumawonetsa kamangidwe kolakwika ka crypt (muvi) ndi ma cell odziwika bwino a goblet (g).Nthenga za fluorescein (f) zimatuluka m'maselo a epithelial, kuwonetsa kuchuluka kwa mitsempha yamagazi.Maselo ambiri otupa (mivi) amawoneka mu lamina propria (lp).Smt.Kanema Wowonjezera 5. (b, d, h, l) Zithunzi zokonzedwa pogwiritsa ntchito LabVIEW 2021.
Zithunzi zofananira za kutupa kwa m'matumbo zimawonetsedwa, kuphatikiza ulcerative colitis (UC) (Chithunzi 4g-i) ndi Crohn's colitis (Chithunzi 4j-l).Kuyankha kotupa kumaganiziridwa kuti kumadziwika ndi zosokoneza za crypt zomwe zimakhala ndi ma cell a goblet otuluka.Fluorescein imafinyidwa m'maselo a epithelial, kuwonetsa kuchuluka kwa mitsempha yamagazi.Maselo ambiri otupa amatha kuwoneka mu lamina propria.
Tawonetsa kagwiritsidwe ntchito kachipatala ka flexible fiber-coupled confocal laser endoscope yomwe imagwiritsa ntchito scanner ya MEMS yokhazikika kuti apeze zithunzi za vivo.Pafupipafupi, mitengo ya chimango mpaka 20 Hz imatha kupezedwa pogwiritsa ntchito kachulukidwe kakang'ono ka Lissajous kuti muchepetse zoyenda.Njira ya kuwala imapindidwa kuti ipereke kukula kwa mtengo ndi kabowo kakang'ono kokwanira kuti akwaniritse kusintha kwa 1.1 µm.Zithunzi za fluorescent za khalidwe la histological zidapezedwa panthawi ya colonoscopy yokhazikika ya colonic mucosa, tubular adenomas, hyperplastic polyps, ulcerative colitis, ndi Crohn's colitis.Maselo amodzi amatha kudziwika, kuphatikiza ma colonocyte, ma cell a goblet, ndi maselo otupa.Zinthu za mucosal monga mapangidwe a crypt, crypt cavities, ndi lamina propria zitha kuzindikirika.Chida cholondola chimapangidwa ndi makina ang'onoang'ono kuti zitsimikizire kulondola kwazinthu zowoneka bwino komanso zamakina mkati mwa 2.4mm mainchesi x 10mm kutalika chida.Maonekedwe a kuwala amachepetsa kutalika kwa nsonga yolimba ya distal mokwanira kuti alole kudutsa molunjika kudzera mu kukula kwake (3.2 mm m'mimba mwake) njira yogwirira ntchito mu endoscopes yachipatala.Choncho, mosasamala kanthu za wopanga, chipangizocho chingagwiritsidwe ntchito kwambiri ndi madokotala pamalo okhala.Chisangalalo chinachitidwa pa λex = 488 nm kuti asangalatse fluorescein, utoto wovomerezeka ndi FDA, kuti mupeze kusiyana kwakukulu.Chidacho chidakonzedwanso popanda zovuta kwa mizere 18 pogwiritsa ntchito njira zovomerezeka zachipatala.
Zopangira zina ziwiri za zida zatsimikiziridwa ndichipatala.Cellvizio (Mauna Kea Technologies) ndi probe-based confocal laser endoscope (pCLE) yomwe imagwiritsa ntchito mtolo wa multimode coherent fiber optic zingwe kusonkhanitsa ndi kutumiza zithunzi za fluorescence1.Kalilore wa galvo yemwe ali pamalo oyambira amajambula kutsogolo kumapeto kwa proximal.Magawo a kuwala amasonkhanitsidwa mu ndege yopingasa (XY) ndi kuya kwa 0 mpaka 70 µm.Ma microprobe kits amapezeka kuchokera ku 0.91 (19 G singano) mpaka 5 mm m'mimba mwake.Chigamulo chotsatira cha 1 mpaka 3.5 µm chinakwaniritsidwa.Zithunzi zidasonkhanitsidwa pamlingo wa 9 mpaka 12 Hz wokhala ndi gawo limodzi lowonekera kuchokera ku 240 mpaka 600 µm.Pulatifomu yakhala ikugwiritsidwa ntchito kuchipatala m'malo osiyanasiyana kuphatikiza ndulu, chikhodzodzo, colon, esophagus, mapapo, ndi kapamba.Optiscan Pty Ltd yapanga endoscope-based confocal laser endoscope (eCLE) yokhala ndi injini yojambulira yomwe imamangidwa mu chubu choyikapo (distal end) ya endoscope yaukadaulo (EC-3870K, Pentax Precision Instruments) 17 .Gawo la kuwala linkagwiritsidwa ntchito pogwiritsa ntchito ulusi wamtundu umodzi, ndipo kusanthula m'mbali kunkagwiritsidwa ntchito pogwiritsa ntchito makina a cantilever kupyolera mu foloko yopangira resonant.A Shape Memory Alloy (Nitinol) actuator amagwiritsidwa ntchito kupanga axial kusamuka.Kutalika konse kwa gawo la confocal ndi 5 mm.Poyang'ana, lens ya GRIN yokhala ndi nambala yobowoleza ya NA = 0.6 imagwiritsidwa ntchito.Zithunzi zopingasa zinapezedwa ndi malingaliro a lateral ndi axial a 0.7 ndi 7 µm, motero, pamlingo wa 0.8-1.6 Hz ndi gawo la 500 µm × 500 µm.
Timawonetsa kusamvana kwa subcellular mu vivo fluorescence imaging imaging kuchokera mthupi la munthu kudzera mu endoscope yachipatala pogwiritsa ntchito scanner ya distal MEMS.Fluorescence imapereka kusiyana kwakukulu kwazithunzi, ndipo ma ligand omwe amamangiriza ku maselo amtundu wa cell amatha kulembedwa ndi fluorophores kuti apereke chidziwitso cha maselo kuti adziwe bwino matenda18.Njira zina zowonera mu vivo microendoscopy zikupangidwanso. OCT imagwiritsa ntchito utali wolumikizana waufupi kuchokera ku gwero la kuwala kwa burodibandi kusonkhanitsa zithunzi mundege yoyima yokhala ndi kuya> 1 mm19. OCT imagwiritsa ntchito utali wolumikizana waufupi kuchokera ku gwero la kuwala kwa burodibandi kusonkhanitsa zithunzi mundege yoyima yokhala ndi kuya> 1 mm19. ОКТ Христа короткую длину когерентности широкополосного источника света для сбора изображений в вертикальной плоскостим 19 >. OCT imagwiritsa ntchito utali waufupi wa gwero la kuwala kwa burodibandi kuti apeze zithunzi mundege yoyima ndi> 1 mm kuya kwa19. 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 м19 вькополосного. OCT imagwiritsa ntchito utali wolumikizana waufupi wa gwero la kuwala kwa burodibandi kuti apeze zithunzi> 1 mm19 mundege yoyima.Komabe, kusiyanitsa kocheperako kumeneku kumadalira kusonkhanitsa kwa kuwala kobwerera m'mbuyo komanso kusasinthika kwazithunzi kumachepetsedwa ndi zinthu zakale.Photoacoustic endoscopy imapanga mu vivo zithunzi kutengera kukula kwamphamvu kwa thermoelastic mu minofu pambuyo pa kuyamwa kwa laser pulse yomwe imapanga mafunde amawu20. Njirayi yawonetsa kuya kwa kujambula> 1 cm m'matumbo amunthu mu vivo kuyang'anira chithandizo. Njirayi yawonetsa kuya kwa kujambula> 1 cm m'matumbo amunthu mu vivo kuyang'anira chithandizo. Этот подход продемонстрировал глубину визуализации > 1 см в толстой кишке человека in vivo для мониторинга терапии. Njirayi yawonetsa kuzama kwa kujambula kwa> 1 cm m'matumbo amunthu mu vivo pakuwunika chithandizo.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 mu vivo для мониторинга терапии. Njirayi yawonetsedwa pakuzama kwazithunzi> 1 cm mumatumbo amunthu mu vivo kuyang'anira chithandizo.Kusiyanitsa kumapangidwa makamaka ndi hemoglobin m'mitsempha.Multiphoton endoscopy imapanga zithunzi za fluorescence zosiyanitsidwa kwambiri pamene zithunzi ziwiri kapena zingapo za NIR zigunda ma biomolecule a minofu nthawi imodzi21. Njirayi imatha kukwaniritsa kuya kwazithunzi> 1 mm ndi kutsika kwa phototoxicity. Njirayi imatha kukwaniritsa kuya kwazithunzi> 1 mm ndi kutsika kwa phototoxicity. Этот подход может обеспечить глубину изображения > 1 мм с низкой фототоксичностью. Njirayi ikhoza kupereka kuya kwa chithunzi> 1 mm ndi phototoxicity yochepa.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. Njirayi ikhoza kupereka kuya kwa chithunzi> 1 mm ndi phototoxicity yochepa.Kuthamanga kwamphamvu kwambiri kwa femtosecond laser pulses kumafunika ndipo njirayi siinatsimikizidwe mwachipatala panthawi ya endoscopy.
Pachitsanzo ichi, sikaniyo imangopatuka kumbuyo, kotero gawo la kuwala limakhala mu ndege yopingasa (XY).Chipangizochi chimatha kugwira ntchito pamlingo wapamwamba kwambiri (20 Hz) kuposa magalasi a galvanic (12 Hz) mu dongosolo la Cellvizio.Wonjezerani kuchuluka kwa mafelemu kuti muchepetse zoyenda ndikuchepetsa mafelemu kuti mukweze chizindikiro.Ma algorithms othamanga kwambiri komanso odzipangira okha amafunikira kuti muchepetse zinthu zazikulu zoyenda zomwe zimayambitsidwa ndi endoscopic motion, kupuma, komanso kuyenda kwamatumbo.Ma parametric resonant scanners awonetsedwa kuti akwaniritsa kusamuka kwa axial mopitilira mazana a ma microns22. Zithunzi zitha kusonkhanitsidwa mu vertical plane (XZ), perpendicular to mucosal surface, kuti apereke malingaliro ofanana ndi a histology (H&E). Zithunzi zitha kusonkhanitsidwa mu vertical plane (XZ), perpendicular to mucosal surface, kuti apereke malingaliro ofanana ndi a histology (H&E). Изображения могут быть получены в вертикальной плоскости (XZ), перпендикулярной поверхности слизистой оболочки, чтобы обезеспе H&E (H&E). Zithunzi zikhoza kutengedwa mu ndege yowongoka (XZ) perpendicular kwa mucosal pamwamba kuti apereke chithunzi chomwecho monga 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 оболочки, чтобы обезеспе гистологическом исследовании (H&E). Zithunzi zitha kujambulidwa mu ndege yowongoka (XZ) yolunjika ku mucosal pamwamba kuti ipereke chithunzi chofanana ndi kuyesa kwa histological (H&E).Chojambuliracho chikhoza kuyikidwa pamalo otsogolera pomwe mtengo wounikira umagwera m'mbali mwa axis yayikulu kuti muchepetse kukhudzidwa kwa ma aberration8.Pafupifupi ma voliyumu ang'onoang'ono ocheperako amatha kupatuka pamawonekedwe akulu.Kusanthula mwachisawawa kutha kuchitidwa kuti apatutse zowunikira kukhala malo omwe amafotokozedwa ndi ogwiritsa9.Gawo la mawonedwe likhoza kuchepetsedwa kuti liwonetsere madera osasinthasintha a chithunzicho, kuwongolera chiŵerengero cha ma signal-to-noise, kusiyana, ndi mawonekedwe.Ma scanner amatha kupangidwa mochuluka pogwiritsa ntchito njira zosavuta.Mazana a zida zitha kupangidwa pa chowotcha chilichonse cha silicon kuti chiwonjezeke kupanga kupanga kotsika mtengo komanso kugawa kwakukulu.
Njira yopindika yowala imachepetsa kukula kwa nsonga yolimba ya distal, kupangitsa kuti ikhale yosavuta kugwiritsa ntchito endoscope ngati chowonjezera panthawi ya colonoscopy.Pazithunzi za fulorosenti zomwe zikuwonetsedwa, mawonekedwe amtundu wa mucosa amatha kusiyanitsa ma tubular adenomas (precancerous) ndi ma polyps a hyperplastic (benign).Zotsatirazi zikuwonetsa kuti endoscopy imatha kuchepetsa kuchuluka kwa ma biopsies osafunikira23.Zovuta zambiri zomwe zimagwirizanitsidwa ndi opaleshoni zimatha kuchepetsedwa, nthawi zowunikira zimatha kukonzedwa, ndipo kusanthula kwa histological kwa zilonda zazing'ono kumatha kuchepetsedwa.Tikuwonetsanso mu vivo zithunzi za odwala omwe ali ndi matenda otupa am'mimba, kuphatikiza ulcerative colitis (UC) ndi Crohn's colitis.Colonoscopy yodziwika bwino yoyera imapereka mawonekedwe owoneka bwino a mucosal pamwamba osakwanitsa kuwunika molondola machiritso a mucosal.Endoscopy angagwiritsidwe ntchito mu vivo kuyesa mphamvu ya mankhwala achilengedwe monga anti-TNF24 antibodies.Kuwunika kolondola kwa vivo kungathenso kuchepetsa kapena kupewa kuyambiranso kwa matenda ndi zovuta monga opaleshoni komanso kukonza moyo wabwino.Palibe zoyipa zoyipa zomwe zidanenedwa m'maphunziro azachipatala okhudzana ndi kugwiritsa ntchito ma endoscope okhala ndi fluorescein mu vivo25. Mphamvu ya laser pa mucosal pamwamba inali yochepa ku <2 mW kuti achepetse chiopsezo cha kuvulala kwa kutentha ndikukwaniritsa zofunikira za FDA pa chiopsezo chosafunikira26 pa 21 CFR 812. Mphamvu ya laser pamtunda wa mucosal inali yochepa ku <2 mW kuti achepetse chiwopsezo cha kuvulala kwamafuta ndikukwaniritsa zofunikira za FDA pazovuta zosafunikira26 pa 21 CFR 812. Momwe mungapangire zolemba pazithunzi zojambulidwa patsamba lawebusayiti <2 мВт, чтобы свести к минимуму риск термического поврежденият FDA ndi kugulitsa но незначительного риска26 согласно 21 CFR 812. Mphamvu ya laser pa mucosal pamwamba inali yochepa ku <2 mW kuti achepetse chiwopsezo cha kuwonongeka kwa kutentha ndikukwaniritsa zofunikira za FDA pangozi yonyozeka26 pansi pa 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 Momwe mungapangire zolemba pazithunzi zapadziko lonse lapansi zomwe zili patsamba <2 мВт, чтобы свести к минимуму риск термического термического повреждения 10 FDA pavреждения 10 FDA 2 относительно незначительного риска26. Mphamvu ya laser pa mucosal pamwamba inali yochepa ku <2 mW kuti achepetse chiwopsezo cha kuwonongeka kwa kutentha ndikukwaniritsa zofunikira za FDA 21 CFR 812 pangozi yosasamala26.
Mapangidwe a chida amatha kusinthidwa kuti apititse patsogolo chithunzithunzi.Ma Optics apadera akupezeka kuti achepetse kutembenuka kozungulira, kukonza mawonekedwe azithunzi ndikuwonjezera mtunda wogwirira ntchito.SIL ikhoza kusinthidwa kuti igwirizane bwino ndi ndondomeko yowonetsera minofu (~ 1.4) kuti igwirizane bwino.Maulendo oyendetsa galimoto amatha kusinthidwa kuti awonjezere mbali yotsalira ya scanner ndikukulitsa mawonekedwe azithunzi.Mutha kugwiritsa ntchito njira zodzichitira nokha kuti muchotse mafelemu azithunzi ndikuyenda kwakukulu kuti muchepetse izi.A field-programmable gate array (FPGA) yokhala ndi data yothamanga kwambiri idzagwiritsidwa ntchito popereka kuwongolera kwanthawi zonse kwanthawi zonse.Kuti zitha kugwiritsidwa ntchito kwambiri pazachipatala, njira zodzipangira zokha ziyenera kukonza zosintha zamagulu ndi zoyenda kuti zitha kutanthauzira zenizeni zenizeni.Makina a monolithic 3-axis parametric resonant scanner atha kukhazikitsidwa kuti ayambitse kusanthula kwa axial 22. Zipangizozi zapangidwa kuti zikwaniritse kusamuka kosasunthika komwe sikunachitikepo> 400 µm posintha ma frequency oyendetsa muulamuliro womwe umakhala ndi kufewetsa kosakanikirana/kuuma kwamphamvu27. Zipangizozi zapangidwa kuti zikwaniritse kusamuka kosasunthika komwe sikunachitikepo> 400 µm posintha ma frequency oyendetsa muulamuliro womwe umakhala ndi kufewetsa kosakanikirana/kuuma kwamphamvu27. Эти устройства были разработаны для достижения беспрецедентного вертикального смещения > 400 мкм путем настройки частоты возбраке возбражение уется смешанной динамикой смягчения/жесткости27. Zipangizozi zapangidwa kuti zikwaniritse kusamuka kosasunthika kosaneneka kwa> 400 µm pokhazikitsa ma frequency oyendetsa munjira yomwe imadziwika ndi zofewa zofewa / zolimba27.這些設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首动频率來实现 4 pa 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 частотиженива смещений й размягчения/затвердевания27. Zipangizozi zidapangidwa kuti zikwaniritse kusamuka kosasunthika komwe sikunachitikepo> 400 µm posintha ma frequency a trigger mu mosakanikirana kufewetsa / kuuma kinetics mode27.M'tsogolomu, kujambula kozungulira koyang'ana kungathandize kuwonetsa khansa yoyambirira (T1a).A capacitive sensing circuit angagwiritsidwe ntchito kufufuza kayendedwe ka scanner ndi kukonza gawo la 28.Kuwongolera gawo lodziyimira pawokha pogwiritsa ntchito sensa yozungulira kumatha kusintha kusintha kwa zida zamanja musanagwiritse ntchito.Kudalirika kwa zida kungawongoleredwe pogwiritsa ntchito njira zodalirika zosindikizira zida kuti muwonjezere kuchuluka kwa makina opangira.Ukadaulo wa MEMS umalonjeza kufulumizitsa kugwiritsa ntchito ma endoscopes powonera epithelium ya ziwalo zopanda kanthu, kuzindikira matenda, ndikuyang'anira chithandizo m'njira yocheperako.Ndi kupititsa patsogolo, njira yatsopano yojambulira iyi ikhoza kukhala njira yotsika mtengo yogwiritsidwa ntchito ngati njira yolumikizirana ndi ma endoscopes azachipatala pakuwunika kwanthawi yayitali ndipo imatha kulowa m'malo mwa kusanthula kwachikhalidwe.
Mayeso a Ray tracing adachitidwa pogwiritsa ntchito pulogalamu ya ZEMAX Optical Design (mtundu wa 2013) kuti adziwe magawo a mawonekedwe owunikira.Mapangidwe apangidwe amaphatikizapo pafupi-diffractive axial resolution, mtunda wogwirira ntchito = 0 µm, ndi gawo lakuwona (FOV) wamkulu kuposa 250 × 250 µm2.Pachisangalalo pa kutalika kwa mawonekedwe λex = 488 nm, fiber single-mode (SMF) idagwiritsidwa ntchito.Mapiritsi a Achromatic amagwiritsidwa ntchito kuchepetsa kusiyana kwa kusonkhanitsa kwa fluorescence (Chithunzi 5a).Mtengowo umadutsa mu SMF yokhala ndi mainchesi a 3.5 μm ndipo popanda kudumpha umadutsa pakati pa chowunikiracho ndi mainchesi a 50 μm.Gwiritsani ntchito mandala omiza olimba (hemispherical) okhala ndi cholozera chowoneka bwino (n = 2.03) kuti muchepetse kutsika kozungulira ndikuwonetsetsa kukhudzana kwathunthu ndi mucous membrane.Kuyika kwa optics kumapereka chiwerengero chonse cha NA = 0.41, pamene NA = nsinα, n ndi ndondomeko yowonongeka ya minofu, α ndiye mlingo waukulu wa convergence angle.Ma diffraction-limited lateral ndi axial resolutions ndi 0.44 ndi 6.65 µm, motsatana, pogwiritsa ntchito NA = 0.41, λ = 488 nm, ndi n = 1.3313.Magalasi omwe amapezeka pamalonda okha ndi awiri akunja (OD) ≤ 2 mm ankaganiziridwa.Njira yowonekera imapindidwa, ndipo mtengo womwe umachoka pa SMF umadutsa pakatikati pa chojambuliracho ndipo umayang'aniridwa ndi galasi lokhazikika (0.29 mm m'mimba mwake).Kukonzekera uku kufupikitsa kutalika kwa malekezero okhwima kuti atsogolere kupita kwa endoscope kudzera mu njira yogwirira ntchito (3.2 mm m'mimba mwake) ya ma endoscopes azachipatala.Izi zimapangitsa kuti zikhale zosavuta kugwiritsa ntchito ngati chowonjezera panthawi ya endoscopy.
Opindidwa kalozera wowunikira komanso kuyika kwa endoscope.(a) Mtsinje wachisangalalo umatuluka mu OBC ndikudutsa pobowo lapakati la sikani.Mtengowo umakulitsidwa ndikuwonetseredwa kuchokera pagalasi lozungulira lokhazikika kubwereranso ku scanner kuti atembenuke kumbuyo.Ma optics olunjika amakhala ndi ma lens awiri achromatic doublet ndi mandala omiza olimba (hemispherical) omwe amalumikizana ndi mucosal pamwamba.ZEMAX 2013 (https://www.zemax.com/) ya kapangidwe ka kuwala ndi kayesedwe ka ray.(b) Imawonetsa komwe kuli zida zosiyanasiyana, kuphatikiza fiber single mode (SMF), scanner, magalasi, ndi magalasi.Solidworks 2016 (https://www.solidworks.com/) idagwiritsidwa ntchito pakupanga kwa 3D kwa paketi ya endoscope.
SMF (#460HP, Thorlabs) yokhala ndi gawo la gawo la 3.5 µm pamtunda wa 488 nm idagwiritsidwa ntchito ngati "dzenje" pakusefa kwapang'onopang'ono kwa kuwala kosakhazikika (mkuyu 5b).Ma SMF ali otsekeredwa mu machubu a polima osinthika (#Pebax 72D, Nordson MEDICAL).Kutalika kwa pafupifupi mamita 4 kumagwiritsidwa ntchito kuonetsetsa mtunda wokwanira pakati pa wodwalayo ndi dongosolo lojambula zithunzi.Ma lens awiri a 2 mm MgF2 okhala ndi ma achromatic doublet lens (#65568, #65567, Edmund Optics) ndi ma lens a hemispherical 2 mm (#90858, Edmund Optics) adagwiritsidwa ntchito kuyang'ana mtengowo ndikusonkhanitsa fluorescence.Ikani chubu chomaliza chachitsulo chosapanga dzimbiri (4 mm kutalika, 2.0 mm OD, 1.6 mm ID) pakati pa utomoni ndi chubu chakunja kuti mulekanitse kugwedezeka kwa sikani.Gwiritsani ntchito zomatira zachipatala kuti muteteze chidacho ku madzi am'thupi ndi njira zogwirira ntchito.Gwiritsani ntchito machubu ochepetsa kutentha kuti muteteze zolumikizira.
The compact scanner imapangidwa pa mfundo ya parametric resonance.Ikani pobowo ya 50 µm pakatikati pa chowunikira kuti mutumize mtengo wosangalatsa.Pogwiritsa ntchito ma drive a quadrature comb-driven, mtengo wokulitsidwa umapatutsidwa modutsa njira ya orthogonal (ndege ya XY) mumayendedwe a Lissajous.Bolodi yopezera deta (#DAQ PCI-6115, NI) idagwiritsidwa ntchito kupanga ma analogi kuti aziwongolera sikani.Mphamvu zidaperekedwa ndi amplifier yamagetsi apamwamba (#PDm200, PiezoDrive) kudzera pa mawaya owonda (#B4421241, MWS Wire Industries).Pangani mawaya pa ma electrode armature.Sikinayi imagwira ntchito pafupipafupi pafupi ndi 15 kHz (yothamanga kwambiri) ndi 4 kHz (yoyenda pang'onopang'ono) kuti ikwaniritse FOV mpaka 250 µm × 250 µm.Kanema amatha kuwomberedwa pamlingo wa 10, 16, kapena 20 Hz.Mitengo iyi ya chimango imagwiritsidwa ntchito kuti ifanane ndi kubwereza kwapatani ya Lissajous, zomwe zimatengera mtengo wa X ndi Y ma frequency akisikana29.Tsatanetsatane wa kusinthanitsa pakati pa mtengo wa chimango, kusanja kwa pixel, ndi kachulukidwe kake kawonekedwe zafotokozedwa mu ntchito yathu yapitayi14.
Laser state state (#OBIS 488 LS, coherent) imapereka λex = 488 nm kusangalatsa fluorescein kwa kusiyana kwa zithunzi (Mkuyu 6a).Ma pigtails owoneka amalumikizidwa ndi fyuluta yolumikizira kudzera pa zolumikizira za FC / APC (kutaya 1.82 dB) (Mkuyu 6b).Mtengowo umapatutsidwa ndi galasi la dichroic (#WDM-12P-111-488/500:600, Oz Optics) mu SMF kudzera pa cholumikizira china cha FC/APC.Mogwirizana ndi 21 CFR 812, mphamvu yochitika ku minofu imakhala yochepera 2 mW kuti ikwaniritse zofunikira za FDA pachiwopsezo chonyozeka.Fluorescence idadutsa pagalasi la dichroic ndi zosefera zazitali (#BLP01-488R, Semrock).Fluorescence idatumizidwa ku chowunikira cha photomultiplier chubu (PMT) (#H7422-40, Hamamatsu) kudzera pa cholumikizira cha FC/PC chogwiritsa ntchito ~ 1 m utali wa fiber multimode wokhala ndi mainchesi 50 µm pakati.Ma siginecha a fluorescent adakulitsidwa ndi amplifier yothamanga kwambiri (#59-179, Edmund Optics).Mapulogalamu apadera (LabVIEW 2021, NI) apangidwa kuti apeze deta yeniyeni komanso kukonza zithunzi.Mphamvu ya laser ndi PMT zosintha zimatsimikiziridwa ndi microcontroller (#Arduino UNO, Arduino) pogwiritsa ntchito bolodi losindikizidwa lapadera.Ma SMF ndi mawaya amatha pazolumikizira ndikulumikizana ndi ma doko a fiber optic (F) ndi mawaya (W) pamalo oyambira (Chithunzi 6c).Makina ojambulira ali pangolo yonyamula (Chithunzi 6d). Transformer yodzipatula idagwiritsidwa ntchito kuchepetsa kutayikira kwapano kukhala <500 μA. Transformer yodzipatula idagwiritsidwa ntchito kuchepetsa kutayikira kwapano kukhala <500 μA. Для ограничения тока утечки до <500 мкА использовался изолирующий трансформатор. Transformer yodzipatula idagwiritsidwa ntchito kuchepetsa kutayikira kwapano kukhala <500 µA.使用隔离变压器将泄漏电流限制在<500 μA. <500μA. Используйте изолирующий трансформатор, чтобы ограничить ток утечки до <500 мкА. Gwiritsani ntchito thiransifoma yodzipatula kuti muchepetse kutayikira kwapano ku <500µA.
dongosolo lowonera.(a) PMT, laser ndi amplifier zili pamalo oyambira.(b) Mu banki ya fyuluta, laser (buluu) ikuyendetsa pa chingwe cha fiber optic kudzera pa cholumikizira cha FC/APC.Mtengowo umapatutsidwa ndi galasi la dichroic (DM) kukhala fiber single mode (SMF) kudzera pa cholumikizira chachiwiri cha FC/APC.Fluorescence (green) imadutsa mu DM ndi long pass filter (LPF) kupita ku PMT kudzera mu multimode fiber (MMF).(c) Mapeto oyandikira a endoscope amalumikizidwa ndi madoko a fiber optic (F) ndi mawaya (W) a siteshoni yoyambira.(d) Endoscope, monitor, base station, kompyuta, ndi kudzipatula thiransifoma pa ngolo yonyamula.(a, c) Solidworks 2016 idagwiritsidwa ntchito pojambula 3D yamakina ojambulira ndi zigawo za endoscope.
Kusunthika kwa lateral ndi axial kwa optics yoyang'ana kunayezedwa kuchokera kumalo kufalikira kwa ma microspheres a fulorosenti (#F8803, Thermo Fisher Scientific) 0.1 µm m'mimba mwake.Sonkhanitsani zithunzi pomasulira ma microspheres mopingasa komanso molunjika mu masitepe 1 µm pogwiritsa ntchito siteji ya mzere (# M-562-XYZ, DM-13, Newport).Zithunzi zojambulidwa pogwiritsa ntchito ImageJ2 kuti mupeze zithunzi zamagulu ang'onoang'ono.
Mapulogalamu apadera (LabVIEW 2021, NI) apangidwa kuti apeze deta yeniyeni komanso kukonza zithunzi.Pa mkuyu.7 ikuwonetsa mwachidule machitidwe omwe amagwiritsidwa ntchito poyendetsa.Mawonekedwe a ogwiritsa ntchito amakhala ndi data acquisition (DAQ), main panel ndi controller panel.Gulu losonkhanitsira deta limalumikizana ndi gulu lalikulu kuti lisonkhanitse ndi kusunga deta yaiwisi, kupereka zolowera pazokonda zosonkhanitsira deta, ndikuyang'anira zosintha za driver wa scanner.Gulu lalikulu limalola wogwiritsa ntchito kusankha masinthidwe omwe akufuna kuti agwiritse ntchito endoscope, kuphatikiza chizindikiro chowongolera scanner, kuchuluka kwazithunzi zamakanema, ndi magawo ogulira.Gululi limathandizanso wogwiritsa ntchito kuwonetsa ndikuwongolera kuwala ndi kusiyana kwa chithunzicho.Pogwiritsa ntchito deta yaiwisi monga kulowetsa, ndondomekoyi imawerengetsera kupindula kwabwino kwa PMT ndikusintha chigawochi pogwiritsa ntchito njira yowonetsera ndemanga (PI)16.Gulu lowongolera limalumikizana ndi bolodi yayikulu ndi bolodi yopezera deta kuti ilamulire mphamvu ya laser ndi kupindula kwa PMT.
Zomangamanga zamapulogalamu adongosolo.Mawonekedwe a ogwiritsa ntchito amakhala ndi ma module (1) data acquisition (DAQ), (2) main panel ndi (3) controller panel.Mapulogalamuwa amayendera limodzi ndikulankhulana kudzera pamizere ya mauthenga.Chinsinsi ndi MEMS: Microelectromechanical System, TDMS: Technical Data Control Flow, PI: Proportional Integral, PMT: Photomultiplier.Mafayilo azithunzi ndi makanema amasungidwa mumitundu ya BMP ndi AVI, motsatana.
Algorithm yowongolera gawo imagwiritsidwa ntchito kuwerengera kuchuluka kwa ma pixel azithunzi pamagawo osiyanasiyana kuti mudziwe mtengo wokwanira womwe umagwiritsidwa ntchito kukulitsa chithunzicho.Pakuwongolera nthawi yeniyeni, gawo la scan scan ndi ± 2.86 ° ndi gawo lalikulu la 0.286 ° kuti muchepetse nthawi yowerengera.Kuphatikiza apo, kugwiritsa ntchito magawo azithunzi okhala ndi zitsanzo zochepa kumachepetsanso nthawi yowerengera chithunzi kuchokera masekondi 7.5 (1 Msample) mpaka masekondi 1.88 (250 Ksample) pa 10 Hz.Magawo olowa awa adasankhidwa kuti apereke chithunzi chokwanira chokhala ndi latency yochepa panthawi ya kujambula kwa vivo.Zithunzi ndi makanema apamoyo amajambulidwa mumitundu ya BMP ndi AVI, motsatana.Zomwe zasungidwa zimasungidwa mu Technical Data Management Flow Format (TMDS).
Pambuyo pokonza zithunzi za mu vivo kuti ziwongolere bwino ndi LabVIEW 2021. Kulondola kumakhala kochepa mukamagwiritsa ntchito njira zowongolera magawo panthawi ya kujambula kwa vivo chifukwa cha nthawi yayitali yowerengera yofunikira.Magawo azithunzi okha ndi nambala zachitsanzo zimagwiritsidwa ntchito.Kuphatikiza apo, ma aligorivimu sagwira ntchito bwino pazithunzi zokhala ndi zinthu zoyenda kapena kusiyanitsa kochepa ndipo zimatsogolera ku zolakwika zowerengera gawo30.Mafelemu pawokha okhala ndi kusiyanitsa kwakukulu komanso zopanda zoyenda adasankhidwa pamanja kuti akonzenso gawo limodzi ndi masikanidwe agawo a ±0.75° mu masitepe 0.01°.Malo onse azithunzi adagwiritsidwa ntchito (mwachitsanzo, 1 Msample ya chithunzi chojambulidwa pa 10 Hz).Table S2 imafotokoza za magawo azithunzi omwe amagwiritsidwa ntchito munthawi yeniyeni komanso pambuyo pokonza.Pambuyo pokonza gawo, fyuluta yapakati imagwiritsidwa ntchito kuchepetsa phokoso lazithunzi.Kuwala ndi kusiyanitsa kumapangidwanso bwino ndi histogram kutambasula ndi kukonza gamma31.
Mayesero azachipatala adavomerezedwa ndi Michigan Medical Institutions Review Board ndipo adachitidwa mu dipatimenti ya Medical Procedures.Kafukufukuyu adalembetsedwa pa intaneti ndi ClinicalTrials.gov (NCT03220711, tsiku lolembetsa: 07/18/2017).Njira zophatikizira zidaphatikizapo odwala (azaka 18 mpaka 100) omwe ali ndi colonoscopy yosankhidwa kale, chiwopsezo chowonjezeka cha khansa yapakhungu, komanso mbiri ya matenda otupa amatumbo.Chilolezo chodziwitsidwa chinapezedwa kuchokera ku phunziro lirilonse lomwe linavomereza kutenga nawo mbali.Njira zosiyanitsira anali odwala omwe anali ndi pakati, omwe anali ndi hypersensitivity ku fluorescein, kapena anali kulandira mankhwala amphamvu a chemotherapy kapena ma radiation.Kafukufukuyu adaphatikizapo odwala otsatizana omwe amakonzekera colonoscopy nthawi zonse ndipo anali woimira Michigan Medical Center.Phunzirolo linachitidwa motsatira Declaration of Helsinki.
Opaleshoni isanachitike, yesani endoscope pogwiritsa ntchito mikanda ya fulorosenti 10 µm (#F8836, Thermo Fisher Scientific) yoikidwa mu nkhungu za silikoni.Silicone sealant (#RTV108, Momentive) idatsanuliridwa mu nkhungu yapulasitiki ya 3D yosindikizidwa 8 cm3.Ikani mikanda ya fulorosenti yamadzi pamwamba pa silicone ndikusiya mpaka madzi auma.
M'matumbo onse adawunikidwa pogwiritsa ntchito colonoscope yachipatala (Olympus, CF-HQ190L) yokhala ndi kuwala koyera.Pambuyo endoscopist watsimikiza dera la matenda akuti, malo osambitsidwa ndi 5-10 ml ya 5% asidi asidi, ndiyeno ndi madzi wosabala kuchotsa ntchofu ndi zinyalala.Mlingo wa 5 ml wa 5 mg/ml fluorescein (Alcon, Fluorescite) anabayidwa kudzera m'mitsempha kapena kupopera pamwamba pa mucosa pogwiritsa ntchito cannula wamba (M00530860, Boston Scientific) yomwe idadutsa munjira yogwirira ntchito.
Gwiritsani ntchito mthirira kuti muchotse utoto wochulukirapo kapena zinyalala kuchokera ku mucosal pamwamba.Chotsani catheter ya nebulizing ndikudutsa endoscope kudzera munjira yogwirira ntchito kuti mupeze zithunzi za ante-mortem.Gwiritsani ntchito chiwongolero cha endoscopic chamitundu yonse kuti muyike nsonga yakutali pamalo omwe mukufuna. Nthawi yonse yomwe imagwiritsidwa ntchito kusonkhanitsa zithunzi za confocal inali <10 min. Nthawi yonse yomwe imagwiritsidwa ntchito kusonkhanitsa zithunzi za confocal inali <10 min. Общее время, затраченное на сбор конфокальных изображений, составило <10 мин. Nthawi yonse yomwe idatengedwa kuti mutenge zithunzi zolumikizana inali <10 min.Nthawi yonse yopeza zithunzi zolumikizana inali yosakwana mphindi 10.Kanema wa Endoscopic White Light adakonzedwa pogwiritsa ntchito makina ojambulira a Olympus EVIS EXERA III (CLV-190) ndikujambulidwa pogwiritsa ntchito chojambulira cha Elgato HD.Gwiritsani ntchito LabVIEW 2021 kujambula ndi kusunga mavidiyo a endoscopy.Kujambula kumalizidwa, endoscope imachotsedwa ndipo minofu yomwe iyenera kuwonedwa imadulidwa pogwiritsa ntchito mphamvu za biopsy kapena msampha. Minofuyo idakonzedwa kuti ipange mbiri yakale (H&E), ndikuwunikidwa ndi katswiri wa GI pathologist (HDA). Minofuyo idakonzedwa kuti ipange mbiri yakale (H&E), ndikuwunikidwa ndi katswiri wa GI pathologist (HDA). Ткани были обработаны для обычной гистологии (H&E) ndi оценены экспертом-патологом желудочно-кишечного тракта (HDA). Minofu idakonzedwa kuti ipange mbiri yakale (H&E) ndikuwunikiridwa ndi katswiri wazotenda zam'mimba (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) ndi оценены экспертом-патологом желудочно-кишечного тракта (HDA). Minofu idakonzedwa kuti ipange mbiri yakale (H&E) ndikuwunikiridwa ndi katswiri wazotenda zam'mimba (HDA).Mawonekedwe a fluorescein adatsimikiziridwa pogwiritsa ntchito spectrometer (USB2000+, Ocean Optics) monga momwe tawonetsera pa Chithunzi S2.
Endoscopes ndi chosawilitsidwa aliyense ntchito ndi anthu (mkuyu 8).Njira zoyeretsera zidachitika motsogozedwa ndi kuvomerezedwa ndi dipatimenti yoyang'anira matenda ndi matenda a Epidemiology ku Michigan Medical Center ndi Central Sterile Processing Unit. Phunziroli lisanachitike, zidazo zidayesedwa ndikuvomerezedwa kuti zithetsedwe ndi Advanced Sterilization Products (ASP, Johnson & Johnson), bungwe lazamalonda lomwe limapereka ntchito zopewera matenda komanso zotsimikizira zoletsa. Phunziroli lisanachitike, zidazo zidayesedwa ndikuvomerezedwa kuti zithetsedwe ndi Advanced Sterilization Products (ASP, Johnson & Johnson), bungwe lazamalonda lomwe limapereka ntchito zopewera matenda komanso zotsimikizira zoletsa. Перед исследованием инструменты были протестированы ndi одобрены для стерилизации компанией Advanced Sterilization Products (ASP, Johnson & Johnson), коммерческой ордгайской, ndi профилактике инфекций ndi проверке стерилизации. Asanaphunzire, zida zidayesedwa ndikuvomerezedwa kuti ziletsedwe ndi Advanced Sterilization Products (ASP, Johnson & Johnson), bungwe lazamalonda lomwe limapereka ntchito zopewera matenda ndi kutsimikizira kuletsa. Перед исследованием инструменты были стерилизованы ndi проверены Advanced Sterilization Products (ASP, Johnson & Johnson), коммерческой организацией, которая предостакестакестанский предостаке предоставные й ndi проверке стерилизации. Zida zidayimitsidwa ndikuwunikiridwa asanaphunzire ndi Advanced Sterilization Products (ASP, Johnson & Johnson), bungwe lazamalonda lomwe limapereka ntchito zopewera matenda ndi kutsimikizira kutsekereza.
Kubwezeretsanso zida.(a) Ma endoscopes amayikidwa m'mathireyi pambuyo poyezetsa chilichonse pogwiritsa ntchito njira ya STERRAD.(b) Ma SMF ndi mawaya amathetsedwa ndi fiber optic ndi zolumikizira zamagetsi, motero, zomwe zimatsekedwa musanakonzenso.
Tsukani ma endoscopes pochita izi: (1) pukutani endoscope ndi nsalu yopanda lint yoviikidwa mu enzymatic cleaner kuyambira proximal mpaka distal;(2) Kumiza chida mu enzymatic detergent solution kwa mphindi zitatu ndi madzi.nsalu yopanda lint.Zolumikizira zamagetsi ndi fiber optic zimaphimbidwa ndikuchotsedwa munjira;(3) Endoscope imakulungidwa ndikuyikidwa mu thireyi ya zida kuti atsekeretse pogwiritsa ntchito STERRAD 100NX, hydrogen peroxide gas plasma.kutentha kwambiri ndi chilengedwe chochepa cha chinyezi.
Zolemba zomwe zagwiritsidwa ntchito ndi/kapena zowunikidwa mu kafukufuku wapano zikupezeka kuchokera kwa olemba omwe afunsidwa pazofunikira.
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