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Tare da haɓaka sabbin kayan laushi masu laushi don na'urorin likitanci da aikace-aikacen likitanci, cikakkiyar siffa ta kayan aikinsu na zahiri da na inji yana da mahimmanci kuma mai wahala.An yi amfani da dabarar nanoindentation da aka gyara (AFM) don siffanta mafi ƙarancin yanayi na sabon lehfilcon A biomimetic silicone hydrogel ruwan tabarau mai rufi tare da Layer na tsarin goga na polymer mai rassa.Wannan hanyar tana ba da damar tantance madaidaicin wuraren tuntuɓar juna ba tare da tasirin extrusion mai ɗanɗano ba lokacin da ke gabatowa da ƙwayoyin polymers.Bugu da ƙari, yana ba da damar sanin halayen injiniya na abubuwan goga guda ɗaya ba tare da tasirin poroelasticity ba.Ana samun wannan ta hanyar zaɓin bincike na AFM tare da ƙira (girman tip, geometry da kuma bazara) wanda ya dace musamman don auna kayan kayan laushi da samfurori na halitta.Wannan hanyar tana haɓaka hankali da daidaito don ingantacciyar ma'auni na kayan laushi mai laushi lehfilcon A, wanda ke da ƙarancin ƙarancin elasticity a saman yanki (har zuwa 2 kPa) da kuma babban elasticity na ciki (kusan 100%) yanayin ruwa mai ruwa. .Sakamakon binciken saman ba wai kawai ya bayyana kaddarorin ultra-laushi na lens na lehfilcon A ba, amma kuma ya nuna cewa ma'aunin goge-goge na polymer mai rassa ya yi daidai da na siliki-hydrogen substrate.Ana iya amfani da wannan dabarar siffa ta saman ga sauran kayan aiki masu laushi da na'urorin likitanci.
Abubuwan injiniyoyi na kayan da aka tsara don hulɗar kai tsaye tare da nama mai rai galibi ana ƙaddara su ta hanyar yanayin halitta.Madaidaicin madaidaicin waɗannan kaddarorin kayan yana taimakawa don cimma halaye na asibiti da ake so na kayan ba tare da haifar da mummunan martani na salon salula ba1,2,3.Don yawancin kayan kama-da-wane, halayen kayan aikin injiniya yana da sauƙin sauƙi saboda samun daidaitattun hanyoyin da hanyoyin gwaji (misali, microindentation4,5,6).Koyaya, don kayan laushi masu laushi irin su gels, hydrogels, biopolymers, sel masu rai, da sauransu, waɗannan hanyoyin gwajin gabaɗaya ba su da amfani saboda iyakokin ƙudurin aunawa da rashin daidaituwa na wasu kayan7.A cikin shekarun da suka wuce, an gyare-gyaren hanyoyin shigar da al'ada na al'ada kuma an daidaita su don kwatanta nau'ikan kayan laushi masu yawa, amma hanyoyi da yawa har yanzu suna fama da mummunan rauni wanda ke iyakance amfani da su8,9,10,11,12,13.Rashin ƙwararrun hanyoyin gwaji waɗanda za su iya daidai da dogaro da keɓance kaddarorin injiniyoyi na kayan supersoft da yadudduka na sama suna iyakance amfani da su a aikace-aikace daban-daban.
A cikin aikinmu na baya, mun gabatar da ruwan tabarau na lehfilcon A (CL), wani abu mai laushi mai laushi tare da duk kaddarorin saman mai laushi waɗanda aka samo daga yuwuwar ƙirar biomimetic wanda aka yi wahayi daga saman cornea na ido.An ɓullo da wannan biomaterial ta hanyar grafting wani reshe, giciye-linked polymer Layer na poly (2-methacryloyloxyethylphosphorylcholine (MPC)) (PMPC) a kan silicone hydrogel (SiHy) 15 tsara don na'urorin kiwon lafiya bisa.Wannan tsari na grafting yana haifar da shimfiɗa a saman wanda ya ƙunshi tsari mai laushi mai laushi kuma mai ƙarfi sosai.Ayyukanmu na baya sun tabbatar da cewa tsarin biomimetic na lehfilcon A CL yana ba da kaddarorin da suka fi dacewa kamar ingantaccen wetting da rigakafin ƙazanta, ƙara yawan lubricity, da rage ƙwayar ƙwayar cuta da ƙwayar cuta15,16.Bugu da ƙari, amfani da haɓaka wannan kayan aikin biomimetic shima yana ba da shawarar ƙarin haɓaka zuwa wasu na'urorin likitanci.Sabili da haka, yana da mahimmanci don siffanta abubuwan da ke saman wannan abu mai laushi mai laushi kuma fahimtar hulɗar injiniyarsa tare da ido don ƙirƙirar ingantaccen tushen ilimi don tallafawa ci gaba da aikace-aikace na gaba.Yawancin ruwan tabarau na SiHy na kasuwanci sun ƙunshi cakuɗaɗɗen nau'in polymers na hydrophilic da hydrophobic waɗanda ke samar da sigar kayan aiki iri ɗaya17.An gudanar da bincike da yawa don bincika kayan aikin su ta hanyar amfani da matsi na gargajiya, ƙwanƙwasa da hanyoyin gwajin microindentation18,19,20,21.Koyaya, sabon ƙirar biomimetic na lehfilcon A CL ya sa ya zama nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau’in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in ya ƙirƙira ƙirar ƙirar ƙirar ƙirar ƙirar ƙirar ƙirar ƙirar ƙirar polymer mai rassa wacce ta bambanta da na tushen tushen SiHy.Sabili da haka, yana da matukar wahala a ƙididdige waɗannan kaddarorin daidai ta amfani da na al'ada da hanyoyin shiga.Hanya mai ban sha'awa tana amfani da hanyar gwajin nanoindentation da aka aiwatar a cikin ƙananan ƙwayoyin cuta (AFM), hanyar da aka yi amfani da ita don ƙayyade kayan aikin injiniya na kayan viscoelastic mai laushi irin su kwayoyin halitta da kyallen takarda, da kuma polymers masu laushi22,23,24,25. .,26,27,28,29,30.A cikin nanoindentation na AFM, mahimman abubuwan gwajin nanoindentation an haɗa su tare da sabbin ci gaba a cikin fasahar AFM don samar da haɓakar ma'auni da gwaji na nau'ikan abubuwan supersoft na zahiri31,32,33,34,35,36.Bugu da kari, fasahar tana ba da wasu fa'idodi masu mahimmanci ta hanyar amfani da nau'ikan geometries daban-daban.mai shiga da bincike da yuwuwar gwaji a cikin kafofin watsa labarai na ruwa daban-daban.
AFM nanoindentation za a iya raba yanayin yanayi zuwa manyan sassa uku: (1) kayan aiki (ma'auni, ganowa, bincike, da dai sauransu);(2) ma'aunin ma'auni (kamar ƙarfi, ƙaura, gudu, girman ramp, da sauransu);(3) Gudanar da bayanai (gyara tushe, kimanta ma'anar taɓawa, dacewa da bayanai, ƙirar ƙira, da sauransu).Matsala mai mahimmanci tare da wannan hanya ita ce yawancin karatu a cikin wallafe-wallafen ta yin amfani da rahoton nanoindentation na AFM sosai daban-daban sakamakon ƙididdiga don samfurin / cell / nau'in nau'in nau'in 37,38,39,40,41.Alal misali, Leka et al.An yi nazari tare da kwatanta tasirin tasirin binciken binciken AFM akan ma'auni na Matasa na samfuran samfuran hydrogel masu kama da juna da kuma sel iri-iri.Suna bayar da rahoton cewa ƙimar modules sun dogara sosai akan zaɓin cantilever da sifar tip, tare da mafi girman ƙimar bincike mai siffar pyramid da mafi ƙarancin ƙimar 42 don bincike mai zagaye.Hakanan, Selhuber-Unkel et al.An nuna yadda saurin indenenter, girman mai shiga da kauri na samfuran polyacrylamide (PAAM) ke shafar ma'aunin matashi wanda aka auna ta ACM43 nanoindentation.Wani abin da ke daure kai shi ne rashin daidaitattun kayan gwaji marasa ƙarfi da hanyoyin gwaji kyauta.Wannan yana sa yana da matukar wahala a sami ingantaccen sakamako tare da amincewa.Duk da haka, hanyar tana da amfani sosai don ma'auni na dangi da kuma kimantawa tsakanin nau'in samfurin irin wannan, misali ta yin amfani da nanoindentation na AFM don rarrabe kwayoyin halitta daga kwayoyin cutar kansa 44, 45.
Lokacin gwada kayan laushi tare da nanoindentation na AFM, babban ƙa'idar babban yatsan yatsa shine a yi amfani da bincike tare da ƙaramin ƙarancin bazara (k) wanda ya dace da ma'auni na samfuri da tip na hemispherical / zagaye don kada binciken farko ya huda samfurin saman akan. tuntuɓar farko tare da kayan laushi.Hakanan yana da mahimmanci cewa siginar jujjuyawar da binciken ya samar ya kasance mai ƙarfi sosai don gano shi ta hanyar tsarin gano laser24,34,46,47.A cikin yanayin sel daban-daban masu laushi masu laushi, kyallen takarda da gels, wani ƙalubale shine a shawo kan ƙarfin mannewa tsakanin binciken da saman samfurin don tabbatar da ma'auni mai ƙima kuma abin dogaro48,49,50.Har zuwa kwanan nan, yawancin aiki akan nanoindentation na AFM ya mayar da hankali kan nazarin halayen injiniya na kwayoyin halitta, kyallen takarda, gels, hydrogels, da biomolecules ta amfani da ƙananan bincike mai zurfi, wanda ake kira colloidal probes (CPs).. kamar carbon (DLC).Kodayake CP-AFM nanoindentation sau da yawa shine zaɓi na farko don ƙirar samfurin taushi, yana da matsalolin kansa da iyakancewa.Yin amfani da manyan na'urori masu girman micron-spherical yana haɓaka jimlar lamba na tip tare da samfurin kuma yana haifar da babban asarar ƙudurin sararin samaniya.Don samfurori masu laushi, marasa daidaituwa, inda kayan aikin injiniya na abubuwan gida zasu iya bambanta da yawa daga matsakaita akan yanki mai faɗi, CP indentation na iya ɓoye duk wani rashin daidaituwa a cikin kaddarorin akan sikelin gida52.Ana yin bincike-bincike na colloidal ta hanyar haɗe sassan kolloidal masu girman micron zuwa cantilevers maras ƙarfi ta amfani da adhesives na epoxy.Tsarin masana'anta da kansa yana cike da matsaloli da yawa kuma yana iya haifar da rashin daidaituwa a cikin tsarin tantancewar bincike.Bugu da kari, girma da taro na colloidal barbashi kai tsaye suna shafar babban ma'auni na daidaitawa na cantilever, kamar mitar resonant, taurin bazara, da karkatar da hankali56,57,58.Don haka, hanyoyin da aka saba amfani da su don bincike na AFM na al'ada, irin su zafin jiki na zafin jiki, bazai samar da daidaitaccen daidaituwa ga CP ba, kuma ana iya buƙatar wasu hanyoyin don yin waɗannan gyare-gyare57, 59, 60, 61. Gwajin shigar da CP na yau da kullum yana amfani da manyan karkatattun cantilever zuwa nazarin kaddarorin samfurori masu laushi, wanda ke haifar da wata matsala lokacin da aka daidaita halin da ba a kai ba na cantilever a ƙananan ƙananan ƙetare62,63,64.Hanyoyin shigar da colloidal na zamani yawanci suna la'akari da juzu'i na cantilever da aka yi amfani da su don daidaita binciken, amma watsi da tasirin ƙwayoyin colloidal, wanda ke haifar da ƙarin rashin tabbas a cikin daidaiton hanyar38,61.Hakazalika, na roba moduli da aka lasafta ta hanyar dacewa da samfurin lamba suna dogara kai tsaye ga lissafin lissafi na bincike na indentation, kuma rashin daidaituwa tsakanin tip da samfurin saman halaye na iya haifar da rashin kuskure27, 65, 66, 67, 68. Wasu ayyukan kwanan nan ta Spencer et al.Abubuwan da ya kamata a yi la'akari da su lokacin da aka kwatanta da gogewar polymer mai laushi ta amfani da hanyar nanoindentation na CP-AFM.Sun bayar da rahoton cewa riƙewar ruwa mai danko a cikin gogayen polymer a matsayin aikin saurin yana haifar da haɓakar hawan kai kuma saboda haka ma'auni daban-daban na abubuwan dogaro da sauri30,69,70,71.
A cikin wannan binciken, mun siffata yanayin shimfidar wuri na ultra-soft sosai na roba kayan lehfilcon A CL ta amfani da hanyar AFM nanoindentation da aka gyara.Idan aka ba da kaddarorin da sabon tsarin wannan abu, ƙimar hankali na hanyar indentation na gargajiya a fili bai isa ba don siffanta yanayin wannan abu mai laushi mai laushi, don haka ya zama dole a yi amfani da hanyar nanoindentation na AFM tare da mafi girman hankali da ƙananan hankali.matakin.Bayan nazarin gazawar da matsalolin da ake samu na colloidal AFM probe nanoindentation dabaru, mun nuna dalilin da ya sa muka zabi wani karami, al'ada-tsara AFM bincike don kawar da hankali, baya amo, ma'anar lamba na lamba, ma'auni gudun modules na taushi iri-iri kayan kamar ruwa riƙewa. dogara.kuma daidai gwargwado.Bugu da ƙari, mun sami damar auna daidai siffa da girman tip ɗin shigarwa, yana ba mu damar yin amfani da ƙirar mazugi mai dacewa don ƙayyade ma'auni na elasticity ba tare da tantance wurin lamba na tip tare da kayan ba.Zato guda biyu a fakaice waɗanda aka ƙididdige su a cikin wannan aikin sune cikakkun kaddarorin kayan roba da maɗaukakin maɗaukaki mai zurfi mai zaman kansa.Ta amfani da wannan hanyar, mun fara gwada ma'auni masu laushi masu laushi tare da sanannen modules don ƙididdige hanyar, sannan mu yi amfani da wannan hanyar don siffata saman kayan ruwan tabarau daban-daban guda biyu.Ana sa ran wannan hanyar keɓance saman nanoindentation na AFM tare da ƙarin hankali ana tsammanin za a iya amfani da su ga nau'ikan kayan aikin ultrasoft iri-iri na biomimetic tare da yuwuwar amfani a cikin na'urorin likitanci da aikace-aikacen ilimin halitta.
Lehfilcon A tuntuɓar ruwan tabarau (Alcon, Fort Worth, Texas, Amurka) da silicone hydrogel substrates an zaɓi don gwajin nanoindentation.An yi amfani da hawan ruwan tabarau na musamman a gwajin.Don shigar da ruwan tabarau don gwaji, an sanya shi a hankali a kan tsayawar mai siffar kubba, tabbatar da cewa babu kumfa na iska ya shiga ciki, sannan a gyara shi tare da gefuna.Wani rami a cikin abin da ke saman abin riƙe ruwan tabarau yana ba da dama ga cibiyar gani na ruwan tabarau don gwaje-gwajen nanoindentation yayin riƙe da ruwa a wurin.Wannan yana sa ruwan tabarau su cika ruwa.An yi amfani da 500 μl na maganin marufi na ruwan tabarau azaman maganin gwaji.Don tabbatar da ƙididdigan sakamakon, kasuwancin da ba a kunna polyacrylamide (PAAM) hydrogels an shirya su daga polyacrylamide-co-methylene-bisacrylamide abun da ke ciki (100 mm Petrisoft Petri jita-jita, Matrigen, Irvine, CA, Amurka), sanannen modulus na roba na 1. kPa.Yi amfani da 4-5 saukad (kimanin 125 µl) na phosphate buffered saline (PBS daga Corning Life Sciences, Tewkesbury, MA, USA) da 1 digo na OPTI-FREE Puremoist ruwan tabarau bayani (Alcon, Vaud, TX, Amurka).) a AFM hydrogel-probe interface.
Samfurori na Lehfilcon A CL da SiHy an hango su ta amfani da tsarin FEI Quanta 250 Field Emission Scanning Electron Microscope (FEG SEM) sanye da na'urar gano na'urar daukar hoto ta Scanning Electron Microscope (STEM).Don shirya samfurori, an fara wanke ruwan tabarau da ruwa kuma a yanka a cikin nau'i mai siffar kek.Don cimma bambancin bambanci tsakanin abubuwan hydrophilic da hydrophobic na samfurori, an yi amfani da 0.10% daidaitawar bayani na RuO4 a matsayin rini, wanda aka yi amfani da samfurori na 30 min.Lehfilcon A CL RuO4 tabo yana da mahimmanci ba kawai don cimma ingantacciyar bambance-bambance ba, amma kuma yana taimakawa wajen adana tsarin ɓangarorin polymer mai rassa a cikin nau'in asali na asali, wanda ake iya gani akan hotunan STEM.An wanke su kuma an bushe su a cikin jerin abubuwan haɗin ethanol / ruwa tare da ƙara yawan ƙwayar ethanol.Sannan an jefa samfuran tare da EMBed 812/Araldite epoxy, wanda ya warke cikin dare a 70°C.Samfurin tubalan da aka samu ta hanyar resin polymerization an yanke su tare da ultramicrome, kuma sakamakon siraran sassan an hango su tare da na'urar ganowa ta STEM a cikin yanayin ƙarancin motsi a saurin ƙarfin lantarki na 30 kV.An yi amfani da tsarin SEM iri ɗaya don cikakken sifa na binciken PFQNM-LC-A-CAL AFM (Bruker Nano, Santa Barbara, CA, Amurka).Hotunan SEM na binciken AFM an samo su a cikin yanayin yanayi mai girma na al'ada tare da haɓaka ƙarfin lantarki na 30 kV.Nemi hotuna a kusurwoyi daban-daban da girma don yin rikodin duk cikakkun bayanai na siffa da girman tip binciken AFM.An auna duk girman kaifin sha'awa a cikin hotuna ta lambobi.
A Dimension FastScan Bio Icon atomic force microscope (Bruker Nano, Santa Barbara, CA, USA) tare da yanayin "PeakForce QNM in Fluid" an yi amfani da shi don hangowa da nanoindentate lehfilcon A CL, SiHy substrate, da samfuran hydrogel PAAm.Don gwaje-gwajen hoto, an yi amfani da binciken PEAKFORCE-HIRS-FA (Bruker) tare da radius mai ƙima na 1 nm don ɗaukar manyan hotuna na samfurin a ƙimar sikanin 0.50 Hz.An ɗauki duk hotuna a cikin bayani mai ruwa.
An gudanar da gwaje-gwajen nanoindentation na AFM ta amfani da bincike na PFQNM-LC-A-CAL (Bruker).Binciken AFM yana da tip siliki akan kauri mai nitride 345 nm, tsayi 54 µm da faɗin 4.5 µm tare da mitar 45 kHz.An ƙera shi musamman don ƙididdigewa da aiwatar da ma'aunin nanomechanical masu ƙididdigewa akan samfuran halitta masu taushi.Ana daidaita na'urori masu auna firikwensin daidaiku a masana'anta tare da saitunan bazara da aka riga aka daidaita.Matsalolin bazara na binciken da aka yi amfani da su a cikin wannan binciken sun kasance a cikin kewayon 0.05-0.1 N/m.Don ƙayyade ainihin siffar da girman tip, an kwatanta binciken daki-daki ta amfani da SEM.A kan fig.Hoto 1a yana nuna babban ƙudiri, ƙaramin ƙarar ƙarar sikanin lantarki na binciken PFQNM-LC-A-CAL, yana ba da cikakkiyar ra'ayi na ƙirar binciken.A kan fig.1b yana nuna girman ra'ayi na saman tip ɗin bincike, yana ba da bayanai game da siffar da girman tip.A matsananciyar ƙarshen, allura ita ce hemisphere game da 140 nm a diamita (Fig. 1c).Ƙarƙashin wannan, tip ɗin ya shiga cikin siffar conical, ya kai tsawon ma'auni na kusan 500 nm.A waje da yankin tapering, tip ɗin yana da siliki kuma yana ƙarewa a cikin jimlar tsayin tip na 1.18 µm.Wannan shine babban ɓangaren aikin tip ɗin bincike.Bugu da ƙari, an yi amfani da babban binciken polystyrene (PS) mai girma (Novascan Technologies, Inc., Boone, Iowa, Amurka) mai tsayin diamita na 45 µm da madaidaicin bazara na 2 N/m don gwaji azaman bincike na colloidal.tare da PFQNM-LC-A-CAL 140 nm bincike don kwatanta.
An ba da rahoton cewa ana iya kama ruwa a tsakanin binciken AFM da tsarin goga na polymer a lokacin nanoindentation, wanda zai yi amfani da karfi a kan binciken AFM kafin ya taɓa saman69.Wannan tasirin extrusion na danko saboda riƙewar ruwa na iya canza zahirin wurin tuntuɓar, don haka yana shafar ma'aunin ma'auni na saman.Don nazarin tasirin jumloli na bincike da saurin shigar da ruwa akan riƙe ruwa, an ƙirƙira magudanar ƙarfi don samfuran lehfilcon A CL ta amfani da binciken diamita na nm 140 a ƙimar ƙaura na 1 µm/s da 2 μm/s.diamita bincike 45 µm, kafaffen saitin ƙarfi 6 nN da aka samu a 1 µm/s.Gwaje-gwaje tare da bincike na 140 nm a diamita an gudanar da su a cikin saurin shigarwa na 1 µm / s da ƙarfin saiti na 300 pN, wanda aka zaɓa don ƙirƙirar matsin lamba a cikin kewayon ilimin lissafi (1-8 kPa) na fatar ido na sama.matsa lamba 72. Soft shirye-sanya samfurori na PAA hydrogel tare da matsa lamba na 1 kPa an gwada don ƙarfin indentation na 50 pN a gudun 1 μm / s ta amfani da bincike tare da diamita na 140 nm.
Tunda tsayin ɓangaren juzu'i na tip na binciken PFQNM-LC-A-CAL yana da kusan 500 nm, ga kowane zurfin zurfin ciki <500nm ana iya ɗauka cikin aminci cewa joometry na binciken a lokacin shigarwa zai kasance da gaskiya ga sa. siffar mazugi.Bugu da ƙari, an ɗauka cewa saman kayan da ke ƙarƙashin gwajin zai nuna wani amsa mai canzawa, wanda kuma za a tabbatar da shi a cikin sassan da ke gaba.Sabili da haka, dangane da nau'i da girman tip, mun zaɓi samfurin madaidaicin mazugi wanda Briscoe, Sebastian da Adams suka kirkira, wanda ke samuwa a cikin software na mai siyarwa, don aiwatar da gwaje-gwajen nanoindentation na AFM (NanoScope).Software na nazarin bayanai na rabuwa, Bruker) 73. Samfurin yana kwatanta dangantakar tilastawa F(δ) don mazugi tare da lahani koli.A kan fig.Hoto na 2 yana nuna ma'aunin lissafi a yayin hulɗar mazugi mai ƙarfi tare da tip mai siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar siffar, a cikin radius radius , b shine radius a ƙarshen spherical tip, δ. lamba radius.zurfin zurfin ciki, θ shine rabin kusurwar mazugi.Hoton SEM na wannan binciken yana nuna a sarari cewa diamita na 140nm diamita yana haɗuwa ta atomatik zuwa mazugi, don haka a nan b ana bayyana shi ta hanyar R kawai, watau b = R cos θ.Software da aka kawo mai siyarwa yana ba da alaƙar mazugi don ƙididdige ƙididdige ƙimar Matasa (E) daga bayanan rarrabuwar ƙarfi da ɗauka a > b.Dangantaka:
inda F shine ƙarfin shigar, E shine ma'aunin Matasa, ν shine rabon Poisson.Ana iya ƙididdige radiyon tuntuɓar a ta amfani da:
Tsarin juzu'i na lamba na mazugi mai ƙarfi tare da tip mai siffa mai matsewa a cikin kayan lens ɗin tuntuɓar Lefilcon tare da saman Layer na goge-goge na polymer mai rassa.
Idan ≤ b, alaƙar tana raguwa zuwa ma'auni don mai sikeli na al'ada;
Mun yi imanin cewa hulɗar binciken indenting tare da tsarin reshe na PMPC polymer brush zai sa lamba ta radius a ya zama mafi girma fiye da radius lamba b.Don haka, don duk ma'aunin ƙididdigewa na ma'auni na roba da aka yi a cikin wannan binciken, mun yi amfani da dogaro da aka samu don yanayin a> b.
Kayan aikin biomimetic na ultrasoft da aka yi nazari a cikin wannan binciken an zana su gaba ɗaya ta amfani da sikanin watsawa na lantarki (STEM) na sashin giciye na samfurin giciye da microscopy na atomatik (AFM) na saman.An yi wannan cikakken bayanin yanayin saman azaman haɓaka aikin mu da aka buga a baya, wanda muka ƙaddara cewa tsarin goga mai ƙarfi na polymeric na PMPC-gyara lehfilcon A CL surface ya nuna irin kayan aikin injiniya zuwa nama na asali na asali 14.Don haka, muna komawa zuwa saman ruwan tabarau kamar kayan biomimetic14.A kan fig.3a,b yana nuna sassan giciye na tsarin goga na PMPC polymer mai rassa akan saman wani yanki na lehfilcon A CL da substrate SiHy wanda ba a kula da shi ba, bi da bi.An kara nazarin abubuwan da aka yi amfani da su ta hanyar amfani da hotuna na AFM masu girma, wanda ya kara tabbatar da sakamakon binciken STEM (Fig. 3c, d).Haɗe tare, waɗannan hotuna suna ba da kimanin tsawon tsarin goga mai rahusa na PMPC a 300-400 nm, wanda ke da mahimmanci don fassara ma'aunin nanoindentation na AFM.Wani maɓalli mai mahimmanci da aka samo daga hotunan shine cewa gabaɗayan tsarin saman kayan CL biomimetic ya bambanta da na kayan aikin SiHy.Wannan bambance-bambance a cikin ilimin halittar jikinsu na iya bayyanawa yayin hulɗar injiniyarsu tare da binciken AFM mai indenting kuma daga baya a cikin ƙididdige ƙimar modul.
Hotunan STEM na tsaka-tsaki na (a) lehfilcon A CL da (b) SiHy substrate.Sikelin bar, 500 nm.Hotunan AFM na saman dafilcon A CL substrate (c) da tushe na SiHy (d) (3 µm × 3 µm).
Polymers da aka yi wahayi da sifofin goga na polymer suna da taushi a zahiri kuma an yi nazari sosai kuma an yi amfani da su a aikace-aikacen likitanci daban-daban74,75,76,77.Sabili da haka, yana da mahimmanci a yi amfani da hanyar nanoindentation na AFM, wanda zai iya daidai da kuma dogara auna kayan aikin injiniya.Amma a lokaci guda, keɓaɓɓen kaddarorin waɗannan abubuwa masu laushi masu laushi, irin su ƙarancin ƙarancin ƙarfi, babban abun ciki na ruwa da elasticity mai yawa, galibi suna yin wahalar zaɓar kayan da ya dace, siffa da sifar binciken indenting.girman.Wannan yana da mahimmanci don kada mai haɗawa ya huda ƙasa mai laushi na samfurin, wanda zai haifar da kurakurai wajen ƙayyade ma'anar lamba tare da farfajiyar da yankin lamba.
Don wannan, cikakkiyar fahimta game da ilimin halittar jiki na kayan biomimetic masu laushi masu laushi (lehfilcon A CL) yana da mahimmanci.Bayani game da girman da tsarin branched goge polymer goge ya samu amfani da hanyar da ke samar da tushen fasahar injina ta amfani da dabarun AFM Nanoind.Maimakon micron-spherical colloidal probes, mun zaɓi PFQNM-LC-A-CAL silicon nitride probe (Bruker) tare da diamita na 140 nm, musamman an ƙera shi don ƙididdige taswirar kayan injiniya na samfuran halitta 78, 79, 80 , 81, 82, 83, 84 Dalili na yin amfani da ingantattun bincike idan aka kwatanta da na al'ada colloidal bincike za a iya bayyana ta tsarin fasali na kayan.Kwatanta girman tip na bincike (~ 140 nm) tare da gogewar polymer mai rassa a saman CL lehfilcon A, wanda aka nuna a cikin siffa 3a, ana iya ƙarasa da cewa tip ɗin ya isa ya shiga cikin hulɗar kai tsaye tare da waɗannan sifofin goga, waɗanda yana rage yiwuwar huda tip ta hanyar su.Don kwatanta wannan batu, a cikin siffa 4 shine hoton STEM na lehfilcon A CL da ƙwanƙwasa tip na binciken AFM (wanda aka zana zuwa sikelin).
Tsarin tsari yana nuna hoton STEM na lehfilcon A CL da binciken shigar da ACM (wanda aka zana zuwa sikeli).
Bugu da ƙari, girman tip na 140 nm yana da ƙananan isa don kauce wa hadarin kowane sakamako na extrusion mai ɗorewa a baya da aka ruwaito don gogaggen polymer da aka samar ta hanyar CP-AFM nanoindentation 69,71.Muna ɗauka cewa saboda nau'in mazugi na musamman da kuma ƙananan ƙananan girman wannan tip na AFM (Fig. 1), yanayin ƙarfin ƙarfin da aka yi ta hanyar lehfilcon A CL nanoindentation ba zai dogara da saurin shigarwa ba ko saurin saukewa / saukewa. .Saboda haka, ba a shafar tasirin poroelastic ba.Don gwada wannan hasashe, samfuran lehfilcon A CL an ƙirƙira su a ƙayyadaddun ƙarfi ta amfani da bincike na PFQNM-LC-A-CAL, amma a cikin hanyoyi guda biyu daban-daban, kuma an yi amfani da sakamakon jujjuyawar ƙarfi da jujjuyawar ƙarfi don tsara ƙarfin (nN) Ana nunawa a cikin rabuwa (µm) a cikin hoto na 5a.A bayyane yake cewa ƙarfin ƙarfi a lokacin lodawa da saukewa ya mamaye gaba ɗaya, kuma babu wata bayyananniyar shaida cewa ƙarfin ƙarfi a zurfin zurfin sifili yana ƙaruwa tare da saurin shigarwa a cikin adadi, yana nuna cewa abubuwan goga guda ɗaya sun kasance suna da alaƙa ba tare da tasirin poroelastic ba.Sabanin haka, tasirin riƙewar ruwa (fitarwa na viscous da tasirin poroelasticity) sun bayyana ga binciken 45 µm diamita na AFM a daidai wannan saurin indentation kuma ana nuna su ta hanyar hysteresis tsakanin shimfiɗa da ja da baya, kamar yadda aka nuna a cikin Hoto 5b.Waɗannan sakamakon suna goyan bayan hasashe kuma suna ba da shawarar cewa binciken diamita na 140 nm zaɓi ne mai kyau don siffanta irin waɗannan filaye masu laushi.
lehfilcon A CL indentation karfi masu lankwasa ta amfani da ACM;(a) ta yin amfani da bincike tare da diamita na 140 nm a farashin kaya guda biyu, yana nuna rashin tasirin poroelastic yayin shigar da ƙasa;(b) Yin amfani da bincike tare da diamita na 45 µm da 140 nm.s yana nuna tasirin extrusion danko da poroelasticity don manyan bincike idan aka kwatanta da ƙananan bincike.
Don siffanta saman ultrasoft, hanyoyin nanoindentation na AFM dole ne su sami mafi kyawun bincike don nazarin kaddarorin abubuwan da ake nazari.Bugu da ƙari ga siffar tip da girman girman, ƙaddamar da tsarin tsarin ganowa na AFM, da hankali ga ƙaddamarwa a cikin yanayin gwaji, da kuma kullun cantilever suna taka muhimmiyar rawa wajen ƙayyade daidaito da amincin nanoindentation.ma'auni.Don tsarin mu na AFM, Matsakaicin Mai gano Mahimmanci (PSD) na ganowa shine kusan 0.5 mV kuma ya dogara ne akan ƙimar bazara da aka riga aka daidaita da ƙididdige ƙimar juzu'in juzu'i na binciken PFQNM-LC-A-CAL, wanda yayi daidai da ka'idar load hankali.kasa da 0.1 pN.Sabili da haka, wannan hanyar tana ba da damar auna mafi ƙarancin ƙarfin shigarwa ≤ 0.1 pN ba tare da wani ɓangaren amo ba.Duk da haka, yana da kusan ba zai yiwu ba ga tsarin AFM don rage yawan hayaniyar da ke kewaye da wannan matakin saboda dalilai irin su girgizar injiniya da haɓakar ruwa.Waɗannan abubuwan suna iyakance ƙimar gaba ɗaya na hanyar nanoindentation na AFM kuma suna haifar da siginar amo na baya na kusan ≤ 10 pN.Don haɓakar yanayin ƙasa, samfuran lehfilcon A CL da SiHy an haɗa su a ƙarƙashin cikakken yanayin ruwa ta amfani da bincike na 140 nm don halayyar SEM, kuma sakamakon ƙarfin ƙarfin da aka sanya tsakanin ƙarfi (pN) da matsa lamba.Ana nuna makircin rabuwa (µm) a hoto na 6a.Idan aka kwatanta da madaidaicin tushe na SiHy, lanfilcon A CL mai ƙarfi yana nuna a sarari yanayin tsaka-tsaki yana farawa daga wurin tuntuɓar goga mai yatsu na polymer kuma yana ƙarewa tare da canji mai kaifi a cikin alamar alamar gangara na tip tare da kayan da ke ƙasa.Wannan juzu'in juzu'i na lanƙwan ƙarfi yana ba da haske na gaske na roba goga na polymer mai rassa a saman, kamar yadda aka tabbatar ta hanyar matsawa kusa da bin yanayin tashin hankali da bambanci a cikin kayan aikin injiniya tsakanin tsarin goga da babban kayan SiHy.Lokacin kwatanta lefilcon.Rabuwar matsakaicin tsayin goga na polymer mai rassa a cikin hoton STEM na PCS (Fig. 3a) da ƙarfin ƙarfinsa tare da abscissa a cikin siffa 3a.6a yana nuna cewa hanyar tana iya gano tip da polymer reshe da ke kaiwa saman saman.Tuntuɓi tsakanin tsarin goga.Bugu da kari, kusancin madaidaicin magudanar karfi yana nuna babu tasirin rikon ruwa.A wannan yanayin, babu cikakkiyar mannewa tsakanin allura da saman samfurin.Mafi girman sassan jujjuyawar ƙarfi don samfuran samfuran biyu sun mamaye, suna nuna kamanni na kayan aikin injiniyan kayan aikin.
(a) AFM nanoindentation karfi masu lankwasa don lehfilcon A CL substrates da SiHy substrates, (b) ƙarfin ƙarfin da ke nuna ƙimar lamba ta amfani da hanyar ƙofar amo ta bango.
Don yin nazarin cikakkun bayanai na ƙayyadaddun ƙayyadaddun ƙaƙƙarfan ƙaƙƙarfan ƙarfi, an sake tsara ma'aunin tashin hankali na lehfilcon A CL samfurin a cikin Fig. 6b tare da iyakar ƙarfin 50 pN tare da y-axis.Wannan jadawali yana ba da mahimman bayanai game da hayaniyar bangon asali.Amo yana cikin kewayon ± 10 pN, wanda aka yi amfani da shi don ƙayyade daidai wurin lamba da ƙididdige zurfin shiga.Kamar yadda aka ruwaito a cikin wallafe-wallafen, gano wuraren tuntuɓar yana da mahimmanci don tantance daidaitattun kaddarorin kayan kamar modulus85.Hanyar da ta haɗa da sarrafa atomatik na bayanan lanƙwan ƙarfi ya nuna ingantaccen dacewa tsakanin daidaita bayanai da ma'aunin ƙididdigewa don kayan taushi86.A cikin wannan aikin, zaɓin wuraren tuntuɓar mu abu ne mai sauƙi da manufa, amma yana da iyakokinsa.Hanyar mu na ra'ayin mazan jiya don tantance wurin tuntuɓar na iya haifar da ƙimar ƙimar modul ƙima don ƙaramin zurfin zurfin ciki (<100 nm).Yin amfani da gano abubuwan taɓawa na tushen algorithm da sarrafa bayanai na atomatik na iya zama ci gaba da wannan aikin a nan gaba don ƙara haɓaka hanyarmu.Don haka, don amo na asali na asali akan tsari na ± 10 pN, muna ayyana wurin tuntuɓar a matsayin ma'aunin bayanai na farko akan axis x a cikin Hoto 6b tare da ƙimar ≥10 pN.Sa'an nan, daidai da amo bakin kofa na 10 pN, a tsaye layi a matakin ~ 0.27 µm alama batu na lamba tare da surface, bayan da mikewa kwana ci gaba har sai da substrate hadu da indentation zurfin ~ 270 nm.Abin sha'awa, dangane da girman fasalin goga na polymer mai rassa (300-400 nm) wanda aka auna ta hanyar yin amfani da hanyar hoto, zurfin shigar da CL lehfilcon Samfurin da aka lura ta amfani da hanyar kofa na amo yana kusan 270 nm, wanda yake kusa da shi. girman ma'auni tare da STEM.Waɗannan sakamakon sun ƙara tabbatar da dacewa da kuma amfani da siffa da girman tip ɗin bincike na AFM don shigar da wannan tsari mai laushi mai laushi da na roba mai ƙarfi na polymer.Wannan bayanan kuma yana ba da shaida mai ƙarfi don tallafawa hanyarmu ta yin amfani da hayaniyar baya a matsayin kofa don tantance wuraren tuntuɓar juna.Don haka, duk wani sakamako mai ƙididdigewa da aka samu daga ƙirar lissafin lissafi da dacewa da lanƙwasa ya kamata ya zama daidai.
Ma'auni na ƙididdiga ta hanyoyin AFM nanoindentation sun dogara gaba ɗaya akan ƙirar lissafin da aka yi amfani da su don zaɓin bayanai da bincike na gaba.Sabili da haka, yana da mahimmanci a yi la'akari da duk abubuwan da suka danganci zaɓi na mai shiga, kayan kayan aiki da makanikai na hulɗar su kafin zaɓar wani samfurin.A wannan yanayin, an nuna tipar enometry a hankali ta amfani da kwastomomi masu kyau (Fig.) Na Piereting Tiveet ɗin da ke da kyau don bamban cutar Lehfilcon a CL79 samfurori .Wani muhimmin abu da ya kamata a yi la'akari da shi a hankali shine elasticity na kayan polymer da ake gwadawa.Kodayake bayanan farko na nanoindentation (Figs. 5a da 6a) sun bayyana a fili sifofin da ke tattare da tashin hankali da matsawa, watau, cikakken dawo da kayan aiki na roba, yana da matukar muhimmanci don tabbatar da nau'in nau'i na lambobi. .Don wannan ƙarshen, an yi abubuwan shigar guda biyu a jere a wuri ɗaya a saman samfurin lehfilcon A CL a ƙimar shigar da 1 µm/s ƙarƙashin cikakken yanayin ruwa.Ana nuna sakamakon lanƙwan ƙarfin ƙarfi a cikin fig.7 kuma, kamar yadda ake tsammani, haɓakawa da matsawa na kwafi guda biyu kusan iri ɗaya ne, suna nuna babban elasticity na tsarin goga na polymer mai rassa.
Maɓallan ƙarfin shigar da ƙarfi biyu a wuri ɗaya akan saman lehfilcon A CL suna nuna madaidaicin elasticity na ruwan tabarau.
Dangane da bayanin da aka samu daga hotuna na SEM da STEM na tip na bincike da kuma lehfilcon A CL surface, bi da bi, mazugi-Sphere model ne m mathematics wakilci na hulda tsakanin AFM bincike tip da taushi polymer abu da ake gwada.Bugu da ƙari, don wannan ƙirar mazugi-Sphere, ainihin zato game da kaddarorin roba na kayan da aka buga suna riƙe gaskiya ga wannan sabon abu na biomimetic kuma ana amfani da su don ƙididdige modules na roba.
Bayan cikakken kimantawa na hanyar nanoindentation na AFM da abubuwan da ke tattare da su, gami da kaddarorin binciken indentation (siffa, girman, da tsaurin bazara), hankali (hayaniyar bayan fage da ƙimar lamba), da samfuran dacewa da bayanai (ma'auni na ƙididdigewa), hanyar ta kasance. amfani.Haɓaka samfurori masu laushi masu laushi na kasuwanci don tabbatar da ƙididdiga masu yawa.An gwada polyacrylamide na kasuwanci (PAAM) hydrogel tare da modules na roba na 1 kPa a ƙarƙashin yanayin ruwa ta amfani da bincike na 140 nm.An bayar da cikakkun bayanai game da gwaji da ƙididdiga a cikin Ƙarin Bayani.Sakamakon ya nuna cewa matsakaicin matsakaicin da aka auna ya kasance 0.92 kPa, kuma % RSD da kashi (%) sabawa daga abin da aka sani sun kasance ƙasa da 10%.Waɗannan sakamakon sun tabbatar da daidaito da sake fasalin hanyar AFM nanoindentation da aka yi amfani da su a cikin wannan aikin don auna moduli na kayan ultrasoft.Fuskokin samfuran lehfilcon A CL da SiHy tushe an ƙara siffanta su ta amfani da hanyar AFM nanoindentation iri ɗaya don nazarin madaidaicin lamba na saman ultrasoft a matsayin aikin zurfin ciki.An ƙirƙira maƙallan ɓangarorin ƙarfi na indentation don samfurori guda uku na kowane nau'in (n = 3; indentation ɗaya a kowane samfurin) a ƙarfin 300 pN, saurin 1 µm / s, da cikakken hydration.An kiyasta lanƙwan rabon ƙarfin nitsewa ta amfani da ƙirar mazugi.Don samun modules dangane da zurfin ciki, an saita wani yanki mai faɗin 40nm na ƙarfin ƙarfin a kowane haɓaka na 20nm wanda ya fara daga wurin tuntuɓar, da auna ƙimar ma'aunin a kowane mataki na lanƙwan ƙarfin.Spin Cy et al.An yi amfani da irin wannan hanya don siffanta modul gradient na poly(lauryl methacrylate) (P12MA) polymer brushes ta amfani da colloidal AFM bincike nanoindentation, kuma sun yi daidai da bayanai ta amfani da Hertz lamba model.Wannan tsarin yana ba da maƙasudin madaidaicin lamba (kPa) tare da zurfin indentation (nm), kamar yadda aka nuna a hoto na 8, wanda ke nuna alamar alamar lamba/zurfin gradient.Matsakaicin ƙididdiga na ƙididdiga na CL lehfilcon A samfurin yana cikin kewayon 2-3 kPa a cikin babban 100 nm na samfurin, bayan abin da ya fara karuwa da zurfi.A gefe guda kuma, lokacin da aka gwada madaidaicin tushe na SiHy ba tare da fim mai kama da goga a saman ba, matsakaicin zurfin shigar da aka samu a ƙarfin 300 pN bai wuce 50 nm ba, kuma ƙimar ƙimar da aka samu daga bayanan kusan 400 kPa. , wanda yayi daidai da ƙimar ƙimar Matasa don kayan girma.
Alamun lamba (kPa) vs. zurfin indentation (nm) don lehfilcon A CL da SiHy substrates ta amfani da hanyar nanoindentation na AFM tare da juzu'i na mazugi don auna modulus.
Mafi girman saman sabon tsarin biomimetic mai reshe na goga na polymer yana nuna ƙaramin ƙarancin elasticity (2-3 kPa).Wannan zai dace da ƙarshen rataye kyauta na goga mai cokali mai yatsu kamar yadda aka nuna a hoton STEM.Yayin da akwai wasu shaidun ma'aunin gradient a gefen waje na CL, babban maɗaukakin maɗaukakin maɗaukaki ya fi tasiri.Koyaya, saman 100 nm na saman yana tsakanin 20% na jimlar tsayin goga na polymer mai rassa, don haka yana da kyau a ɗauka cewa ƙimar ƙimar ma'aunin ma'aunin a cikin wannan zurfin zurfin zurfin zurfin zurfin zurfin zurfin zurfin zurfin zurfin 20% ne kuma ba su da ƙarfi sosai. dogara da tasirin abin ƙasa.
Saboda keɓantaccen ƙirar biomimetic na lehfilcon A tuntuɓar ruwan tabarau, wanda ya ƙunshi reshen PMPC polymer goga Tsarin da aka grafted saman saman SiHy substrates, yana da matukar wahala a dogara da ƙayyadaddun kayan aikin injiniya na tsarin su ta amfani da hanyoyin aunawa na gargajiya.Anan mun gabatar da hanyar nanoindentation na AFM na ci gaba don daidaitaccen halayen ultra-taushi kayan kamar lefilcon A tare da babban abun ciki na ruwa da madaidaicin elasticity.Wannan hanya ta dogara ne akan yin amfani da binciken AFM wanda girman tip da lissafi aka zaɓa a hankali don dacewa da ma'auni na tsarin fasali mai laushi da za a buga.Wannan haɗe-haɗe na ma'auni tsakanin bincike da tsari yana ba da ƙarin hankali, yana ba mu damar auna ƙananan modulus da abubuwan da suka dace na roba na abubuwan goga na polymer mai rassa, ba tare da la'akari da tasirin poroelastic ba.Sakamakon ya nuna cewa keɓaɓɓen goge goge na PMPC polymer mai rassa halayyar fuskar ruwan tabarau yana da ƙarancin elasticity na roba (har zuwa 2kPa) da kuma ƙarfin ƙarfi sosai (kusan 100%) lokacin da aka gwada shi a cikin yanayi mai ruwa.Sakamakon nanoindentation na AFM kuma ya ba mu damar siffata alamar alamar lamba / zurfin gradient (30 kPa/200 nm) na saman ruwan tabarau na biomimetic.Wannan gradient na iya zama saboda bambancin modules tsakanin gogashin polima mai rassa da siHy substrate, ko tsarin reshe/yawan gogewar polymer, ko haɗin gwiwa.Koyaya, ana buƙatar ƙarin bincike mai zurfi don cikakken fahimtar alaƙar da ke tsakanin tsari da kaddarorin, musamman tasirin reshen goga akan kaddarorin injina.Makamantan ma'auni na iya taimakawa wajen siffanta kaddarorin injiniyoyi na saman sauran kayan masu laushi da na'urorin likitanci.
Abubuwan da aka ƙirƙira da/ko tantancewa yayin binciken na yanzu suna samuwa daga mawallafa bisa ga buƙata mai ma'ana.
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