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Munguva pfupi yapfuura, makemikari-isina antimicrobial platform yakavakirwa nanotechnology uchishandisa artificial water nanostructures (EWNS) yakagadzirwa. EWNS ine high surface charge uye yakapfuma mu reactive oxygen species (ROS) inogona kudyidzana nekumisa huwandu hwehutachiona, kusanganisira hutachiona hwekudya. Pano zvinoratidzwa kuti zvivakwa zvavo panguva yekubatanidza zvinogona kurongeka uye kugadziridzwa kuti vawedzere kuwedzera simba ravo reantibacterial. Iyo EWNS murabhoritari chikuva chakagadzirirwa kukwenenzvera zvivakwa zveEWNS nekushandura synthesis paramita. Hunhu hweEWNS zvivakwa (charge, size, uye ROS content) yakaitwa pachishandiswa nzira dzemazuva ano dzekuongorora. Pamusoro pezvo, utachiona hwechikafu hwakadai seEscherichia coli, Salmonella enterica, Listeria innocua, Mycobacterium para fortitum, uye Saccharomyces cerevisiae dzakaiswa pamusoro peiyo organic madomasi emuzambiringa kuti aongorore kugona kwavo kusaita hutachiona. Mhedzisiro yakapihwa pano inoratidza kuti zvivakwa zveEWNS zvinogona kurongeka zvakanaka panguva yekubatanidza, zvichikonzera kuwedzera kweexponential mukuita basa rekuita. Kunyanya, mutero wepamusoro wakawedzera nechikamu chechina, uye zvinyorwa zveROS zvakawedzera. Huwandu hwekubvisa hutachiona hwaienderana nehutachiona uye hwakabva pa1.0 kusvika pa3.8 log mushure memaminitsi makumi mana nemashanu ekuratidzwa kune aerosol dose ye40,000 #/cm3 EWNS.
Kusvibiswa kweMicrobial ndiyo honzeri huru yechirwere chekudya chinokonzerwa nekupinzwa kweutachiona kana muchetura wazvo. Chirwere chinokonzerwa nezvokudya chinokonzera zvirwere zvinenge mamiriyoni 76, 325,000 muchipatara, uye 5,000 vanofa gore negore muUnited States moga1. Mukuwedzera, Dhipatimendi Rezvekurima reUnited States (USDA) rinofungidzira kuti kuwedzera kudyiwa kwezvibereko zvinyoro kunokonzera 48 muzana yezvirwere zvose zvinokonzerwa nezvokudya zvinoshumwa muUnited States2. Mutengo wechirwere uye rufu kubva kune hutachiona hwekudya muUnited States wakakwira zvakanyanya, inofungidzirwa neCenters for Disease Control and Prevention (CDC) inodarika US $ 15.6 bhiriyoni pagore3.
Parizvino, kemikari 4, radiation5 uye thermal6 antimicrobial kupindira kuti kuve nechokwadi chekuchengetedzwa kwechikafu kunonyanya kuitwa panguva shoma dzakakosha kudzora nzvimbo (CCPs) mucheni yekugadzira (kazhinji mushure mekukohwa uye/kana panguva yekurongedza) pane kuramba ichiitwa nenzira yekuti zvibereko zvitsva zvinogona kusvibiswa 7. farm-to-table kuenderera mberi. Kukanganisa kushoma uye mutengo.
A nanotechnology-based chemical-free antimicrobial platform ichangobva kugadzirwa kuti iite kuti mabhakitiriya ari pamusoro uye mumhepo achishandisa artificial water nanostructures (EWNS). Nokuda kwekugadzirwa kweEVNS, maitiro maviri akafanana akashandiswa: electrospray uye ionization yemvura (Fig. 1a). EWNS yakamboratidzwa kuve ine yakasarudzika seti yemuviri uye biological zvivakwa8,9,10. EWNS ine avhareji yemaerekitironi gumi pachimiro uye avhareji nanometer saizi ye25 nm (Fig. 1b,c)8,9,10. Mukuwedzera, electron spin resonance (ESR) yakaratidza kuti EWNS ine huwandu hwakawanda hwemhando yeokisijeni inoshandiswa (ROS), kunyanya hydroxyl (OH•) uye superoxide (O2-) radicals (Fig. 1c) 8. EWNS yakaramba iri mumhepo kwenguva yakareba uye inogona kupindirana nehutachiona hwakamiswa mumhepo uye huripo panzvimbo, kuendesa ROS yavo yekubhadhara uye kukonzera microbial inactivation (Fig. 1d). Izvi zvidzidzo zvekare zvakaratidza kuti EWNS inokwanisa kudyidzana pamwe nekumisa mabhakitiriya akasiyana-siyana egiremu-negative uye gram-positive ehutano hwehutano hwevanhu, kusanganisira mycobacteria, pamusoro uye mumhepo8,9. Transmission electron microscopy yakaratidza kuti kusashanda kwakakonzerwa nekuvhiringwa kwesero membrane. Uye zvakare, acute inhalation zvidzidzo zvakaratidza kuti madosi akakwira eEWNS haakonzere kukuvara kwemapapu kana kuzvimba8.
(a) Electrospray inoitika kana yakakwira voltage pakati pecapillary ine mvura uye counter electrode. (b) Kushandiswa kwemagetsi akakwirira kunoguma muzviitiko zviviri zvakasiyana: (i) electrospraying yemvura uye (ii) chizvarwa chezvinoita okisijeni marudzi (ions) akavharirwa muEWNS. (c) Iyo yakasarudzika chimiro cheEWNS. (d) EWNS ine nharembozha nekuda kwechimiro chadzo chenanoscale uye inogona kudyidzana nehutachiona hwemhepo.
Kugona kweEWNS antimicrobial platform yekumisa tupukanana tune chikafu pamusoro pechikafu chitsva chakaratidzirwawo munguva pfupi yapfuura. Izvo zvakare zvakaratidzwa kuti iyo EWNS yepamusoro chargeji inogona kushandiswa pamwe chete nemunda wemagetsi kune yakanangwa kuendesa. Zvinotonyanya kukosha, mhedzisiro yekutanga inovimbisa yeanosvika 1.4 log kuderedzwa kweiyo organic tomato chiitiko kurwisa tupukanana twechikafu seE. coli neListeria yakaonekwa mukati me90 maminetsi ekusangana neEWNS pamwero we50,000#/cm311. Uye zvakare, yekutanga organoleptic evaluation bvunzo yakaratidza hapana organoleptic mhedzisiro kana ichienzaniswa neyekutonga tomato. Kunyangwe iyi yekutanga kusashanda mhinduro inovimbisa kuchengetedzwa kwechikafu kunyangwe pakaderera EWNS madosi e50,000 #/cc. ona, zviri pachena kuti yakakwirira inactivation zvingava zvinobatsira kuwedzera kuderedza njodzi yeutachiona uye kuparara.
Pano, tichatarisa tsvakiridzo yedu pakuvandudzwa kwepuratifomu yechizvarwa cheEWNS kuti tiite zvakanaka maparamita ekutanga uye kukwidziridza physicochemical zvimiro zveEWNS kusimudzira kugona kwavo kweantibacterial. Kunyanya, optimization yakatarisana nekuwedzera yavo yekuchaja yepasi (yekuvandudza yakanangwa kuendesa) uye ROS zvemukati (kunatsiridza inactivation kunyatsoshanda). Hunhu hwe optimized physico-chemical properties (saizi, charge uye ROS content) vachishandisa nzira dzemazuva ano dzekuongorora uye kushandisa tupukanana twekudya twakawanda seE. coli, S. enterica, L. innocua, S. cerevisiae uye M. parafortuitum.
EVNS yakagadzirwa panguva imwe chete ye electrospraying uye ionization yemvura yakachena yakachena (18 MΩ cm-1). Atomizer yemagetsi 12 inowanzo shandiswa kuita atomize zvinwiwa uye synthetic polymer uye ceramic zvimedu 13 uye fiber 14 yehukuru hwakadzorwa.
Sezvakatsanangurwa muzvinyorwa zvakapfuura 8, 9, 10, 11, muchiyedzo chakajairika, mhepo yakakwira inoiswa pakati pesimbi capillary neiyo grounded counter electrode. Munguva iyi, zviitiko zviviri zvakasiyana zvinoitika: 1) electrospray uye 2) ionization yemvura. Munda wemagetsi wakasimba pakati pemaelectrode maviri unokonzera mhosva dzisina kunaka dziuye pamusoro pemvura yakapfupikiswa, zvichikonzera kuumbwa kweTaylor cones. Nekuda kweizvozvo, madonhwe emvura akachajirwa zvakanyanya anoumbwa, ayo anoramba achitsemuka kuita zvidimbu zvidiki, maererano neRayleigh theory16. Panguva imwecheteyo, simba remagetsi rinoita kuti mamwe mamorekuru emvura aparadzanise uye abvise maerekitironi (ionization), nokudaro achigadzira huwandu hwakawanda hwemhando dzeokisijeni (ROS)17. Panguva imwe chete yakagadzirwa ROS18 mapaketi akaiswa muEWNS (Fig. 1c).
Pamusoro pemuonde. 2a inoratidza EWNS chizvarwa system yakagadziridzwa uye yakashandiswa muEWNS synthesis muchidzidzo ichi. Mvura yakacheneswa yakachengetwa mubhodhoro rakavharwa yakadyiswa kuburikidza neTeflon tube (2 mm yemukati dhayamita) kune 30G simbi isina tsvina tsono (metal capillary). Sezvinoratidzwa mumufananidzo 2b, kuyerera kwemvura kunodzorwa nemhepo yemhepo mukati mebhodhoro. Iyo tsono inosungirirwa kune Teflon console iyo inogona kugadziriswa nemaoko kune imwe chinhambwe kubva pakaunda electrode. Kaunda electrode iaruminiyamu dhisiki yakakwenenzverwa ine buri pakati pekuita sampling. Pazasi pekaunda electrode pane aruminiyamu sampling funnel, iyo yakabatana kune yakasara yekuyedza setup kuburikidza nesampling port (Fig. 2b). Zvese sampler zvikamu zvakadzikwa nemagetsi kudzivirira kubhadharisa kuvaka-iyo inogona kukanganisa particle sampling.
(a) Engineered Water Nanostructure Generation System (EWNS). (b) Muchinjikwa chikamu chesampler uye electrospray unit inoratidza iyo inonyanya kukosha paramita. (c) Kuedza kuseta kwebhakitiriya kusashanda.
Iyo EWNS chizvarwa sisitimu inotsanangurwa pamusoro inokwanisa kushandura makiyi ekushandisa paramita kuti ifambise kugadzirisa kwakanaka kweEWNS zvivakwa. Gadzirisa iyo yakashandiswa voltage (V), chinhambwe pakati petsono uye counter electrode (L), uye kuyerera kwemvura (φ) kuburikidza necapillary kuti unyatso-tune maitiro eEWNS. Zviratidzo [V (kV), L (cm)] zvinoshandiswa kuratidza misanganiswa yakasiyana. Gadzirisa kuyerera kwemvura kuti uwane yakagadzikana Taylor koni yeimwe seti [V, L]. Nezvinangwa zvechidzidzo ichi, aperture ye counter electrode (D) yakaiswa pa 0.5 inches (1.29 cm).
Nekuda kweiyo diki geometry uye asymmetry, simba remunda wemagetsi harigone kuverengerwa kubva kumisimboti yekutanga. Pane kudaro, QuickField™ software (Svendborg, Denmark)19 yakashandiswa kuverenga nzvimbo yemagetsi. Munda wemagetsi hausi yunifomu, saka kukosha kwemunda wemagetsi pamucheto we capillary kwakashandiswa seyereferensi kukosha kune akasiyana magadzirirwo.
Munguva yekudzidza, akati wandei musanganiswa wevoltage uye chinhambwe pakati petsono neiyo counter electrode yakaongororwa maererano neTaylor cone kuumbwa, Taylor cone kugadzikana, EWNS kugadzikana kwekugadzira, uye kuberekazve. Musanganiswa wakasiyana-siyana unoratidzwa muSupplementary Tafura S1.
Kubuda kweEWNS chizvarwa system yakabatana zvakananga neScanning Mobility Particle Sizer (SMPS, modhi 3936, TSI, Shoreview, Minnesota) kuyera huwandu hwenhamba uye yakashandiswa neFaraday aerosol electrometer (TSI, modhi 3068B, Shoreview, USA). MN) kuyera kuyerera kweaerosol, sezvakatsanangurwa mune yedu yakambodhindwa9. Ose ari maviri SMPS uye aerosol electrometer yakatorwa pamwero wekuyerera we 0.5 L/min (yakazara sampu inoyerera 1 L/min). Particle concentrations uye aerosol fluxes akayerwa 120 s. Dzokorora kuyerwa kwe30 nguva. Iyo yakazara aerosol charge inoverengerwa kubva pazviyero zvazvino, uye avhareji EWNS charge inofungidzirwa kubva pahuwandu hwehuwandu hweEWNS zvidimbu zvakatorwa. Avhareji mari yeEWNS inogona kuverengerwa uchishandisa Equation (1):
apo IEl ndiyo yakayerwa ikozvino, NSMPS inhamba yenhamba yakayerwa neSMPS, uye φEl inhamba yekuyerera kune electrometer.
Nekuti hunyoro hunyoro (RH) hunokanganisa kuchaja kwepasi, tembiricha uye (RH) zvakachengetwa zvisingaite pa21 ° C uye 45%, zvichiteerana, panguva yekuyedza.
Atomic force microscopy (AFM), Asylum MFP-3D (Asylum Research, Santa Barbara, CA) uye AC260T probe (Olympus, Tokyo, Japan) yakashandiswa kuyera ukuru uye hupenyu hweEWNS. Iyo AFM scan rate ndeye 1 Hz uye nzvimbo ye scan i 5 µm×5 µm ine 256 scan mitsara. Mifananidzo yese yakaiswa pasi pekutanga kurongeka kwemufananidzo uchishandisa Asylum software (mask ine huwandu hwe100 nm uye chikumbaridzo che100 pm).
Bvisa sampling funnel uye isa iyo mica pamusoro pechinhambwe che 2.0 cm kubva pakaunda electrode kweavhareji yenguva ye120 s kudzivirira kubatana kwezvikamu uye kuumbwa kwemadonhwe asina kujairika pane mica pamusoro. EWNS yakashandiswa zvakananga kune ichangobva kuchekwa mica nzvimbo (Ted Pella, Redding, CA). Pakarepo mushure mekuputika, iyo mica pamusoro yakaonekwa uchishandisa AFM. Iyo yekumusoro yekubata kona ye ichangobva kuchekwa isina kugadziridzwa mica iri padyo ne0 °, saka EWNS inopararira pamusoro peiyo mica pamusoro mune domed shape20. Iyo dhayamita (a) uye kureba (h) yemadonhwe anopararira akayerwa zvakananga kubva kuAFM topography uye akashandiswa kuverenga iyo domed diffusion vhoriyamu EWNS tichishandisa nzira yedu yakambosimbiswa8. Tichifunga kuti iyo onboard EVNS ine vhoriyamu yakafanana, dhayamita yakaenzana inogona kuverengerwa kubva kuequation (2):
Zvinoenderana neyakagadzirwa nzira yedu, electron spin resonance (ESR) spin trap yakashandiswa kuona kuvapo kwenguva pfupi-yakararama radical intermediates muEWNS. Aerosols akapfuudzwa nemhinduro ine 235 mM DEPMPO (5-(diethoxyphosphoryl) -5-methyl-1-pyrroline-N-oxide) (Oxis International Inc., Portland, Oregon). Zvose EPR zviyero zvakaitwa uchishandisa Bruker EMX spectrometer (Bruker Instruments Inc. Billerica, MA, USA) uye flat cell arrays. Iyo Acquisit software (Bruker Instruments Inc. Billerica, MA, USA) yakashandiswa kuunganidza uye kuongorora data. The ROS characterization yakaitwa chete seti yemamiriro ekushanda [-6.5 kV, 4.0 cm]. EWNS kutariswa kwakayerwa uchishandisa SMPS mushure mekufunga kurasikirwa kweEWNS mune inokanganisa.
Ozone mazinga akatariswa pachishandiswa 205 Dual Beam Ozone Monitor™ (2B Technologies, Boulder, Co)8,9,10.
Kune ese EWNS zvivakwa, kuyerwa kukosha ndiyo nzira yezviyero, uye chikanganiso chekuyera ndiyo yakajairwa kutsauka. T-test yakaitwa kuenzanisa kukosha kweiyo optimized EWNS hunhu nehukoshi hunoenderana nehwaro EWNS.
Mufananidzo 2c unoratidza yakambogadzirwa uye yakaratidza Electrostatic Precipitation Pass Through System (EPES) inogona kushandiswa kunanga EWNS11 kunzvimbo. EPES inoshandisa EWNS kuchaja pamwe chete netambo yakasimba yemagetsi kuti "inongedza" zvakananga panzvimbo yechinangwa. Tsanangudzo yehurongwa hweEPES inoratidzwa mune imwe nguva yakabudiswa naPyrgiotakis et al.11. Saka, EPES ine 3D yakadhindwa PVC chamber ine tapered magumo ane maviri anoenderana Stainless simbi (304 Stainless simbi, mirror polished) mahwendefa esimbi pakati 15.24 cm kureba. Mapuranga akanga akabatanidzwa kune imwe kunze kwepamusoro-voltage sosi (Bertran 205B-10R, Spellman, Hauppauge, NY), bhodhi rezasi raigara rakanaka uye bhodhi repamusoro raigara rakatsikwa pasi (rakayangarara). Madziro ekamuri akafukidzwa nealuminium foil, iyo inodzikwa nemagetsi kudzivirira kurasikirwa kwechikamu. Kamuri ine gonhi rekurodha rakavharwa rinobvumira nzvimbo dzekuyedza kuti dziiswe pamaraki epurasitiki, ichidzisimudza kubva pandiro yesimbi yepasi kudzivirira kukanganiswa kwemagetsi.
Kubudirira kwekuisa kweEWNS muEPES kwakaverengerwa maererano neprotocol yakambogadzirwa yakatsanangurwa muSupplementary Figure S111.
Sekamuri yekudzora, iyo yechipiri inoyerera kuburikidza necylindrical chamber yakabatana munhevedzano neiyo EPES system uchishandisa yepakati HEPA sefa kubvisa EWNS. Sezvinoratidzwa mufig. 2c, iyo EWNS aerosol yakapombwa kuburikidza nemakamuri maviri akabatana munhevedzano. Iyo sefa pakati pekamuri yekudzora uye EPES inobvisa chero EWNS yasara inokonzeresa tembiricha yakafanana (T), hunyoro hunyoro (RH) uye mazinga eozone.
Hurukuro hunokosha hwekudya hwakawanikwa huchisvibisa zvibereko zvitsva seEscherichia coli (ATCC #27325), fecal indicator, Salmonella enterica (ATCC #53647), chirwere chinokonzerwa nechikafu, Listeria innocua (ATCC #33090), imwe nzira kune pathogenic Listeria monocy. , Saccharomyces cerevisiae (ATCC #4098) seimwe nzira yekushatisa mbiriso, uye Mycobacterium parafortuitous (ATCC #19686) sebhakitiriya rinoramba richirarama yakatengwa kubva kuATCC (Manassas, Virginia).
Tenga mabhokisi ematomatisi emuzambiringa kubva kumusika wako wobva waisa mufiriji pa4°C kusvika washandiswa (kusvika mazuva matatu). Sarudza madomasi kuti uedze saizi imwe, inenge 1/2 inch in diameter.
Maprotocols eincubation, inoculation, exposure and colony counting akatsanangurwa muzvinyorwa zvedu zvakapfuura uye akatsanangurwa zvakadzama muSupplementary Data 11. EWNS performance yakaongororwa nekufumura madomasi akaiswa ku40,000 #/cm3 kwemaminetsi makumi mana nemashanu. Muchidimbu, panguva t = 0 min, madomasi matatu akashandiswa kuongorora tupukanana tukasara. Madomasi matatu akaiswa muEPES uye akaoneswa EWNS pa40,000 #/cc (EWNS pachena madomasi) uye mamwe matatu akaiswa mukamuri yekudzora (control tomate). Hapana rimwe remapoka ematomatisi rakaiswa kune mamwe magadzirirwo. EWNS-yakafumurwa matomatisi uye zvidzoreso zvakabviswa mushure memaminitsi makumi mana neshanu kuongorora mhedzisiro yeEWNS.
Kuedza kwega kwega kwakaitwa mumativi matatu. Kuongororwa kwedata kwakaitwa maererano neprotocol inotsanangurwa muSupplementary Data.
E. coli, Enterobacter, uye L. innocua mabhakitiriya masampuli akazarurwa kuEWNS (45 min, EWNS aerosol concentration 40,000 #/cm3) uye asina kuvharwa akaiswa pellet kuti aongorore maitiro ekusaita. Iyo inonaya yakagadziriswa kwemaawa maviri pakupisa kwekamuri mu 0.1 M sodium cacodylate solution (pH 7.4) ine fixative ye 2.5% glutaraldehyde, 1.25% paraformaldehyde uye 0.03% picric acid. Mushure mekugeza, vakagadziriswa ne 1% osmium tetroxide (OsO4) / 1.5% potassium ferrocyanide (KFeCN6) kwe2 h, yakashambidzwa katatu nemvura uye incubated mu 1% uranyl acetate kwe 1 h, ndokushambidzwa kaviri nemvura. Kutevera kupera mvura mumuviri maminitsi gumi imwe neimwe ye50%, 70%, 90%, 100% doro. Mienzaniso yakabva yaiswa mupropylene oxide kweawa imwe uye yakasvibiswa ne1: 1 musanganiswa wepropylene oxide uye TAAP Epon (Marivac Canada Inc. St. Laurent, CA). Iwo masampuli akaiswa muTAAB Epon uye polymerized pa60 ° C kwemaawa makumi mana nemasere. Iyo yakarapwa granular resin yakachekwa uye yakaonekwa neTEM vachishandisa JEOL 1200EX (JEOL, Tokyo, Japan), yakajairika transmission electron maikorosikopu yakashongedzerwa neAMT 2k CCD kamera (Advanced Microscopy Techniques, Corp., Woburn, MA, USA).
Kuedza kwese kwakaitwa mumatatu. Kwenguva yega yega, kugezeswa kwebhakitiriya kwakaiswa mukati katatu, zvichikonzera huwandu hwemapfumbamwe mapoinzi papoint, avhareji yaishandiswa seiyo hutachiona hwekusangana kwechimwe chipenyu ichocho. Kutsauka kwakashandiswa kwakashandiswa sechikanganiso chekuyera. Mapoinzi ese anoverengwa.
Iyo logarithm yekudzikira kwekusangana kwebhakitiriya ichienzaniswa ne t = 0 min yakaverengerwa uchishandisa inotevera fomula:
apo C0 ndiyo mutsara webhakitiriya mumuenzaniso wekutonga panguva 0 (kureva mushure mokunge nzvimbo yaoma asi isati yaiswa mukamuri) uye Cn ndiyo chiyero chebhakitiriya pamusoro pepamusoro mushure me n maminitsi ekutsvaga.
Kuzvidavirira pakushatiswa kwechisikigo kwebhakitiriya panguva ye45 maminetsi ekuratidzwa, Log-Reduction yakaverengerwawo ichienzaniswa nekutonga pa45 maminetsi sezvinotevera:
Iko Cn ndiko kusangana kwebhakitiriya mumuenzaniso wekutonga panguva n uye Cn-Kudzora ndiko kusanganiswa kwehutachiona hwekutonga panguva n. Dhata inounzwa seyakaderedzwa regi kana ichienzaniswa nekutonga (hapana EWNS kuratidzwa).
Munguva yekudzidza, akati wandei musanganiswa wevoltage uye chinhambwe pakati petsono neiyo counter electrode yakaongororwa maererano neTaylor cone kuumbwa, Taylor cone kugadzikana, EWNS kugadzikana kwekugadzira, uye kuberekazve. Musanganiswa wakasiyana-siyana unoratidzwa muSupplementary Tafura S1. Mhosva mbiri dzakasarudzirwa chidzidzo chakakwana chinoratidza yakagadzikana uye inoberekazve zvivakwa (Taylor cone, EWNS kugadzirwa, uye kugadzikana nekufamba kwenguva). Pamusoro pemuonde. 3 inoratidza mhedzisiro pachaji, saizi uye zvirimo zveROS pamakesi maviri. Mhedzisiro yacho zvakare inopfupikiswa muTafura 1. Nekureva, Mufananidzo 3 uye Tafura 1 inosanganisira zvimiro zveyakagadzirwa kare isina-optimized EWNS8, 9, 10, 11 (baseline-EWNS). Kuverengera kukosha kwechiverengero uchishandisa maviri-tailed t-bvunzo anotsikiswazve muSupplementary Table S2. Mukuwedzera, data yakawedzerwa inosanganisira zvidzidzo pamusoro pekuita kwe counter electrode sampling gole dhayamita (D) uye chinhambwe pakati pe electrode yepasi uye kumuromo kwetsono (L) (Supplementary Figures S2 uye S3).
(a–c) AFM saizi kugovera. (d – f) Mucherechedzo wekuchajisa pamusoro. (g) Hunhu hweROS uye ESR.
Izvo zvakakoshawo kuziva kuti kune ese ari pamusoro apa, akayerwa ionization currents aive muchikamu che2-6 µA, uye mavolts aive muchikamu che -3.8 kusvika -6.5 kV, zvichikonzera kushandiswa kwesimba kweiyi single-terminal EWNS isingasviki 50 mW. . chizvarwa module. Kunyangwe EWNS yakagadzirwa pasi pekumanikidzwa kwakanyanya, mazinga eozone aive akaderera kwazvo, asingambopfuuri 60 ppb.
Supplementary Figure S4 inoratidza akafananidzwa minda yemagetsi ye [-6.5 kV, 4.0 cm] uye [-3.8 kV, 0.5 cm] scenarios, zvichiteerana. Minda maererano nemamiriro ezvinhu [-6.5 kV, 4.0 cm] uye [-3.8 kV, 0.5 cm] inoverengwa se 2 × 105 V / m uye 4.7 × 105 V / m, maererano. Izvi zvinofanirwa kutarisirwa, sezvo chiyero chevoltage kusvika kure chakakwira zvakanyanya mune yechipiri kesi.
Pamusoro pemuonde. 3a,b inoratidza EWNS dhayamita yakayerwa neAFM8. Avhareji yeEWNS madhayamita e[-6.5 kV, 4.0 cm] uye [-3.8 kV, 0.5 cm] akaverengwa se27 nm uye 19 nm, zvichiteerana. Iyo geometric chiyero kutsauka kwekugoverwa kwemakesi [-6.5 kV, 4.0 cm] uye [-3.8 kV, 0.5 masendimita] ndeye 1.41 uye 1.45, zvichiteerana, zvichiratidza kugovera saizi yakamanikana. Zvese zviri zviviri saizi uye geometric standard kutsauka zviri padyo nekutanga-EWNS, iri 25 nm uye 1.41, zvichiteerana. Pamusoro pemuonde. 3c inoratidza kugoverwa kwehukuru hweEWNS yekutanga yakayerwa uchishandisa nzira imwechete pasi pemamiriro akafanana.
Pamusoro pemuonde. 3d, e inoratidza mhedzisiro yekuchaja maitiro. Dhata iavhareji zviyero zve 30 panguva imwe chete zviyero zvekuisa pfungwa (#/cm3) uye zvazvino (I). Ongororo inoratidza kuti avhareji yekuchaja paEWNS i22 ± 6 e- uye 44 ± 6 e- ye [-6.5 kV, 4.0 cm] uye [-3.8 kV, 0.5 cm], zvichiteerana. Zvichienzaniswa neBaseline-EWNS (10 ± 2 e-), mutero wavo wepamusoro wakakwira zvakanyanya, zvakapetwa kaviri pane [-6.5 kV, 4.0 cm] mamiriro uye nekana ye [-3 .8 kV, 0.5 cm]. 3f inoratidza yakakosha EWNS data yekubhadhara.
Kubva paEWNS nhamba dzemepu dzekutarisa (Supplementary Figures S5 neS6), zvinogona kuonekwa kuti [-6.5 kV, 4.0 cm] chiitiko chine nhamba yakakwira zvakanyanya yezvimedu pane [-3.8 kV, 0.5 cm] chiitiko. Izvo zvinofanirwa kucherechedzwa kuti EWNS nhamba yekutarisisa yakatariswa kusvika kumaawa e4 (Supplementary Figures S5 uye S6), uko EWNS chizvarwa kugadzikana kwakaratidza mazinga akaenzana ehuwandu hwehuwandu hwehuwandu mune zvese zviitiko.
Mufananidzo 3g inoratidza EPR spectrum mushure mekutonga (kumashure) kubvisa kune yakagadziridzwa EWNS pa [-6.5 kV, 4.0 cm]. Iyo ROS spectrum inofananidzwa zvakare neyekutanga EWNS mubepa rakamboburitswa. Nhamba yakaverengerwa yeEWNS ichiita ne spin trap ndeye 7.5 × 104 EWNS/s, yakafanana neyakadhindwa yakamboburitswa Baseline-EWNS8. Mucherechedzo weEPR wakaratidza zvakajeka kuvapo kwemhando mbiri dzeROS, apo O2- yakatangira, nepo OH• yaivepo mune zvishoma. Pamusoro pezvo, kuenzanisa kwakananga kwekusimba kwepamusoro kwakaratidza kuti iyo EWNS yakagadziridzwa yaive neiyo yakanyanya ROS yemukati kana ichienzaniswa neyekutanga EWNS.
Pamusoro pemuonde. 4 inoratidza kugona kwekuisa kweEWNS muEPES. Iyo data inopfupikiswa muTafura I uye ichienzaniswa neyekutanga EWNS data. Pamakesi ese eEUNS, deposition yaive pedyo ne100% kunyangwe pamhepo yakaderera ye3.0 kV. Kazhinji, 3.0 kV inokwana kuwana 100% deposition zvisinei nekushanduka kwechaji yepasi. Pasi pemamiriro akafanana, kugona kweiyo Baseline-EWNS kwaingova 56% nekuda kwechaji yakaderera (avhareji gumi maerekitironi paEWNS).
Mufananidzo 5 uye Tafura 2 inopfupisa dhigirii rekusashanda kwezvipembenene zvakaiswa pamusoro pematomatisi mushure mekusangana neanenge 40,000 #/cm3 EWNS kwemaminetsi makumi mana neshanu pasi peiyo optimal scenario [-6.5 kV, 4.0 cm]. Inoculated E. coli uye L. innocua yakaratidza kuderera kukuru kwe 3.8 log mushure memaminitsi e45 ekutsvaga. Pasi pemamiriro ezvinhu akafanana, S. enterica yakaratidza kuderedzwa kwegogi rezasi re 2.2 matanda, nepo S. cerevisiae uye M. parafortuitum yakaratidza 1.0 log kuderedza.
Electron micrographs (Mufananidzo 6) inoratidza kuchinja kwemuviri kunokonzerwa neEWNS muE. coli, Salmonella enterica, uye L. innocua masero anotungamirira pakusashanda. Kudzora mabhakitiriya airatidza intact cell membranes, nepo mabhakitiriya akafumurwa akange akanganisa membrane dzekunze.
Electron microscopic imaging yekudzora uye mabhakitiriya akafumurwa akaburitsa membrane kukuvara.
Iyo data pamusoro peiyo physicochemical properties yeEWNS yakagadziridzwa pamwe chete inoratidza kuti EWNS zvivakwa (pamusoro pechaji uye ROS content) zvakavandudzwa zvakanyanya zvichienzaniswa neyakadhindwa EWNS yekutanga data8,9,10,11. Kune rumwe rutivi, saizi yavo yakaramba iri mumhando ye nanometer, iyo yakafanana zvakanyanya neyakaburitswa mhedzisiro, ichivabvumira kugara mumhepo kwenguva yakareba. Iyo yakacherechedzwa polydispersity inogona kutsanangurwa nekuchinja kwepamusoro pechaji, iyo inotaridza hukuru hweRayleigh mhedzisiro, randomness, uye inogona kubatanidzwa kweEWNS. Zvisinei, sekutsanangurwa kwazvakaitwa naNielsen et al.22, high surface charge inoderedza kubuda kwemhepo kuburikidza nekuwedzera kuwedzera simba repamusoro / kushushikana kwekudonha kwemvura. Dzidziso iyi yakasimbiswa nekuyedza microdroplets22 uye EWNS mune yedu yapfuura kudhindwa8. Kurasikirwa kwemaawa ekuwedzera kunogonawo kukanganisa saizi uye kubatsira mukucherechedzwa saizi kugovera.
Pamusoro pezvo, kubhadharisa kwega kwega kunenge 22-44 e-, zvichienderana nemamiriro ezvinhu, iyo yakanyanya kukwirira kana ichienzaniswa neyekutanga EWNS, ine avhareji yekuchaja ye10 ± 2 maerekitironi pachimiro. Nekudaro, zvinofanirwa kucherechedzwa kuti iyi ndiyo avhareji yekuchaja yeEWNS. Seto et al. Izvo zvakaratidzwa kuti kubhadharisa hakuna yunifomu uye kunotevera log-yakajairika kugovera21. Zvichienzaniswa nebasa redu rekare, kupeta kaviri kubhadharisa kwepasi kunowedzera kaviri kugona kwekuisa muEPES system kusvika kunosvika zana.