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Kubereka kweshiri kunoenderana nekukwanisa kwadzo kuchengeta mbeu inogoneka kwenguva yakareba mumasero ekuchengetera urume (SST).Iyo chaiyo nzira iyo spermatozoa inopinda nayo, inogara mukati, uye inosiya SST inoramba iine nharo.Urume hwehuku dzesharkasi hwakaratidza hutsika hwekusangana, huchigadzira nhare dzefilamentous bundle dzine maseru akawanda.Nekuda kwekuomerwa kwekutarisa kufamba uye maitiro espermatozoa mune opaque fallopian chubhu, takashandisa microfluidic mudziyo ine microchannel cross-section yakafanana neye spermatozoa kudzidza spermatozoa agglutination uye motility.Chidzidzo ichi chinokurukura kuti mabundle eurume anoumbwa sei, mafambiro aanoita, uye basa rawo rinobvira mukuwedzera kugara kwembeu muSST.Takaongorora velocity yesperm uye rheological maitiro apo kuyerera kwemvura kwakagadzirwa mukati meiyo microfluidic chiteshi ne hydrostatic pressure (yekuyerera = 33 µm/s).Iyo spermatozoa inowanzoshambira inopesana nezvino (positive rheology) uye velocity ye spermatozoon bundle yakaderedzwa zvakanyanya kana ichienzaniswa nespermatozoa imwe chete.Maspem bundles akaonekwa achifamba ari spiral uye achiwedzera kureba nekukora sezvo mamwe masperm ega achitorwa. Maspem bundles akaonekwa achiswedera uye akanamatira kumadziro emadziro e microfluidic channels kudzivirira kutsvairwa ne fluid flow velocity > 33 µm/s. Maspem bundles akaonekwa achiswedera uye akanamatira kumadziro emadziro e microfluidic channels kudzivirira kutsvairwa ne fluid flow velocity > 33 µm/s. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрофлюидных каналов, чтобы избения 30 смета мкм / p. Maspem bundles akacherechedzwa kuti aswedere uye anamira kumadziro emadziro enzira dzemicrofluidic kudzivirira kutsvairwa pamvura inoyerera> 33 µm/s.观察到精子束接近并粘附在微流体通道的侧壁上，以避免被流体流速> 33 µm/s 扫过.33 µm/s 扫过. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрожидкостного канала, чтобы избежать компание 3 приближаются и прилипают к боковым стенкам микрожидкостного канала, чтобы избежать сместо 3 км/с. Maspem bundles akacherechedzwa kuti aswedere uye anamira kumadziro epadivi eiyo microfluidic chiteshi kudzivirira kukukurwa nekuyerera kwemvura pa> 33 µm/s.Kuongorora uye kutapurirana maelectron microscopy kwakaratidza kuti mabundle esperm aitsigirwa neakawanda dense material.Iyo data yakawanikwa inoratidza kufamba kwakasiyana kweSharkazi huku spermatozoa, pamwe nekugona kwespermatozoa kuagglutinate uye kuumba nharembozha, izvo zvinobatsira kunzwisisa zviri nani kuchengetwa kwenguva refu kwespermatozoa muSMT.
Kuti ibereke muvanhu nemhuka zhinji, urume nemazai zvinofanira kusvika panzvimbo inoberekeswa panguva yakakodzera.Nokudaro, kusangana kunofanira kuitika kusati kwasvika kana panguva ye ovulation.Kune rumwe rutivi, dzimwe mhuka dzinoyamwisa, dzakadai sembwa, pamwe chete nemarudzi asiri mammalia, akadai sezvipembenene, hove, zvinokambaira, uye shiri, zvinochengeta urume munhengo dzazvo dzokubereka nadzo kwenguva yakareba kusvikira mazai azvo agadzirira kubatanidzwa ( asynchronous fertilization 1 ).Shiri dzinokwanisa kuchengetedza hupenyu hwespermatozoa inokwanisa kuisa mazai kwemavhiki maviri-102.
Ichi chinhu chakasiyana chinosiyanisa shiri kubva kune dzimwe mhuka, sezvo ichipa mukana wakakura wekusangana mushure mekuita kamwe chete kwemavhiki akati wandei pasina kusangana panguva imwe chete uye ovulation.Iyo huru yekuchengetera urume nhengo, inodaidzwa kuti sperm storage tubule (SST), iri mumukati memucosal folds panosangana uterovaginal.Kusvika pari zvino, nzira dzekuti urume hupinde, hugare, uye huchibuda mubhangi rembeu hadzisi kunyatsonzwisiswa.Kubva pane zvidzidzo zvekare, mafungiro akawanda akaiswa mberi, asi hapana chimwe chazvo chakasimbiswa.
Forman4 vakafungidzira kuti spermatozoa inochengetedza hugaro hwayo muSST mhango kuburikidza neinoenderera mberi inofamba ichipesana negwara rekuyerera kwemvura kuburikidza nemaprotein channels ari paSST epithelial masero (rheology).ATP inopera nekuda kwekugara kuri kuita flagellar kunodiwa kuchengetedza urume muSST lumen uye motility inozopedzisira yadzikira kudzamara urume hwaburitswa mubhangi rembeu nekuyerera kwemvura uye kutanga rwendo rutsva kudzika nechubhu inokwira ye fallopian kuti isanganise urume.Zai (Forman4).Iyi modhi yekuchengetedza urume inotsigirwa nekuonekwa ne immunocytochemistry yeaquaporins 2, 3 uye 9 iripo muSST epithelial cells.Kusvika pari zvino, zvidzidzo zvehuku semen rheology nebasa rayo muSST kuchengetedza, kusarudzwa kwembeu yemukadzi, uye kukwikwidza kwesperm kuri kushomeka.Muhuku, urume hunopinda mukati memukadzi mushure mekusangana kwechisikigo, asi inopfuura 80% yespermatozoa inoburitswa kubva kumukadzi nguva pfupi mushure mekusangana.Izvi zvinoratidza kuti bonde ndiro rinonyanya kusarudzwa urume mushiri.Pamusoro pezvo, zvakataurwa kuti isingasviki 1% yespermatozoa yakasanganiswa mumukadzi inoguma muSSTs2.Mukuiswa kwehuku mukati memukadzi, huwandu hwespermatozoa hunosvika paSST hunowedzera kuwedzera maawa makumi maviri nemana mushure mekuiswa.Kusvika pari zvino, maitiro ekusarudzwa kwembeu panguva iyi haina kujeka, uye kufamba kwembeu kunogona kuita basa rakakosha muSST kutora urume.Nekuda kwemadziro akakora uye akajeka emachubhu e fallopian, zvinonetsa kutarisa zvakananga kufamba kwembeu mumachubhu eshiri.Naizvozvo, isu hatina ruzivo rwekutanga rwekuti sei spermatozoa shanduko kuenda kuSST mushure mekusangana.
Rheology ichangobva kuzivikanwa sechinhu chakakosha chinodzora kutakura kwembeu munhengo dzemammalian.Zvichienderana nekugona kweiyo motile spermatozoa kutama inopesana, Zaferani et al8 vakashandisa corra microfluidic system kugadzika motile spermatozoa kubva kunyorerwa urume samples.Rudzi urwu rwekuronga urume rwakakosha pakurapwa kwekushaya mbereko uye tsvakiridzo yekiriniki, uye inofarirwa pane nzira dzechinyakare dzinotora nguva uye kushanda nesimba uye dzinogona kukanganisa morphology yembeu uye kusimba kwechimiro.Nekudaro, kusvika parizvino, hapana ongororo yakaitwa pamusoro pemhedzisiro yekuvharira kubva kunhengo dzesikarudzi dzehuku pane sperm motility.
Pasinei nemagadzirirwo anochengetedza urume hwakachengetwa muSST, vaongorori vakawanda vakaona kuti mugari spermatozoa agglutinate musoro-kune-musoro muSST yehuku 9, 10, zvihuta 2, uye turkeys 11 kugadzira agglutinated urume masumbu.Vanyori vanoratidza kuti pane hukama pakati peiyi agglutination uye kuchengetwa kwenguva refu kwespermatozoa muSST.
Tingari neLake12 vakashuma kubatana kwakasimba pakati pespermatozoa mugland yehuku inogamuchira urume uye yakabvunza kana avian spermatozoa agglutinate nenzira yakafanana neyemammalian spermatozoa.Vanotenda kuti kubatana kwakadzika pakati peurume muvas deferens kunogona kunge kuri nekuda kwekunetseka kunokonzerwa nekuvapo kwenhamba yakawanda yembeu munzvimbo diki.
Paunenge uchiongorora maitiro espermatozoa pamasiraidhi egirazi akarembera, zviratidzo zvenguva pfupi zveagglutination zvinogona kuoneka, kunyanya kumucheto kwemadonhwe embeu.Zvisinei, agglutination yaiwanzovhiringidzwa nechiito chekutenderera chakabatanidzwa nekufamba kunoramba kuripo, iyo inotsanangura chimiro chenguva pfupi chechiitiko ichi.Vatsvakurudzi vakacherechedzawo kuti kana diluent yakawedzerwa kumurume, akarebesa "shinda-kufanana" maseru akaunganidzwa.
Kuedza kwekutanga kutevedzera spermatozoon kwakaitwa nekubvisa tambo yakatetepa kubva padonhwe rakarembera, izvo zvakakonzera kuti pave nehurume hwakarebera senge vesicle inobuda kubva padonhwe reurume.Iyo spermatozoa yakabva yangomira nenzira yakafanana mukati mevhesi, asi iyo yese unit yakakurumidza kunyangarika nekuda kwekuganhurwa kwe3D.Nokudaro, kuti udzidze spermatozoa agglutination, zvakakosha kutarisa motility uye mufambiro we spermatozoa zvakananga mune imwe yega sperm storage tubules, izvo zvakaoma kuwana.Naizvozvo, zvinodikanwa kugadzira chiridzwa chinotevedzera spermatozoa kutsigira zvidzidzo zvesperm motility uye agglutination maitiro.Brillard et al13 vakashuma kuti avhareji yehurefu hwemachubhu ekuchengetera urume mumahuku akura i400–600 µm, asi mamwe maSST anogona kureba se2000 µm.Mero neOgasawara14 vakapatsanura maseminiferous glands kuita machubhu ekuchengetedza mbeu akakura uye asina kukura, ose ari maviri akaenzana pakureba (~ 500 µm) nehupamhi hwemutsipa (~ 38 µm), asi dhayamita ye lumen ye tubules yaive 56.6 uye 56.6 µm.., zvichiteerana 11.2 μm, zvichiteerana.Muchidzidzo chezvino, takashandisa microfluidic mudziyo une chiteshi saizi ye200 µm × 20 µm (W × H), iyo chikamu chemuchinjiko chiri padyo neiyo yeSST yakasimudzwa.Pamusoro pezvo, takaongorora kufambiswa kwembeu uye hunhu hwekugwinya mukuyerera kwemvura, izvo zvinoenderana nefungidziro yaForeman yekuti fluid inogadzirwa neSST epithelial masero inochengeta urume mulumen nenzira inopesana (rheological).
Chinangwa chechidzidzo ichi chaiva chekukunda matambudziko ekucherechedza spermatozoa motility mu fallopian tube uye kudzivirira kuoma kwekudzidza rheology uye maitiro espermatozoa munzvimbo ine simba.Muchina wemicrofluidic wakashandiswa unogadzira hydrostatic pressure kutevedzera kufamba kwembeu munhengo yesikarudzi yehuku.
Pakadonhedzwa donhwe rembeu yakanyungudutswa (1:40) mumudziyo wemicrochannel, marudzi maviri embeu motility yaigona kuzivikanwa (sperm yakasarudzika uye yakasungwa urume).Mukuwedzera, spermatozoa yaifarira kushambira inopesana nezvino (positive rheology; vhidhiyo 1, 2). Kunyangwe mabhururu eurume aive neakaderera velocity pane iya yembeu yakasurukirwa (p <0.001), yakawedzera chikamu chembeu ichiratidza rheotaxis yakanaka (p <0.001; Tafura 2). Kunyangwe mabhururu eurume aive neakaderera velocity pane iya yembeu yakasurukirwa (p <0.001), yakawedzera chikamu chembeu ichiratidza rheotaxis yakanaka (p <0.001; Tafura 2). Хотя пучки сперматозоидов имели более низкую скорость, чем у одиночных сперматозоидов (p <0,001), они увеличивали проецент, процент, процент, програмя ложительный реотаксис (p <0,001; таблица 2). Kunyange zvazvo spermatozoa mabundles aiva nevelocity yakaderera pane imwe spermatozoa (p <0.001), yakawedzera chikamu che spermatozoa chinoratidza rheotaxis yakanaka (p <0.001; Tafura 2).尽管精子束的速度低于孤独精子的速度（p < 0.001），但它們增加了显示阳性流变性的爾0.表2）.尽管 精子束 的 速度 低于 孤独 的 速度（p <0.001） , 但 增加 显示 阳性 流 炾 流 炾 流 炾 流 炾 流 炾 流 炾 流红 流灵 流灵 流 爱 流 爱 流 流约。 .001 ; 2.......））) Хотя скорость пучков сперматозоидов была ниже, чем у одиночных сперматозоидов (p <0,001), они увеличивали процентозелоий процент спелоий процент спержество (p <0,001). p <0,001; таблица 2). Kunyange zvazvo kukurumidza kwemasumbu eurume kwaive kwakaderera kupfuura kweye spermatozoa imwe chete (p <0.001), yakawedzera chikamu che spermatozoa ne rheology yakanaka (p <0.001; Tafura 2).Positive rheology ye single spermatozoa uye tufts inofungidzirwa kusvika pa53% uye 85%, zvichiteerana.
Izvo zvakaonekwa kuti spermatozoa yehuku sharkasi pakarepo mushure mekubuda kwejaculation inoumba mutsara masumbu, anosanganisira akawanda evanhu.Aya tufts anowedzera pakureba uye kukora nekufamba kwenguva uye anogona kuramba ari mu vitro kwemaawa akati wandei asati aparadza (vhidhiyo 3).Aya mabundle ane filamentous akaita se echidna spermatozoa inoumba pamagumo epididymis.Sharkashi hen semen yakawanikwa iine tsika yakakwira yekuunganidza uye kugadzira bundle reticulate isingasviki miniti imwe mushure mekuunganidzwa.Aya matanda ane simba uye anokwanisa kunamatira kune chero madziro ari pedyo kana zvinhu zvakamira.Kunyangwe masumbu embeu achidzikisa kumhanya kwesero rembeu, zviri pachena kuti macroscopically anowedzera mutsara wawo.Hurefu hwemasumbu hunosiyana zvichienderana nenhamba yembeu yakaunganidzwa mumasumbu.Zvikamu zviviri zvebundle zvakaparadzaniswa: chikamu chekutanga, kusanganisira musoro wakasununguka wembeu yakanyungudutswa, uye chikamu chekupedzisira, kusanganisira muswe uye kumucheto kwese kwese kwembeu.Kushandisa kamera yepamusoro-soro (950 fps), misoro yakasununguka ye spermatozoa yakasvibiswa yakaonekwa muchikamu chekutanga chebhanhire, inotarisira kufamba kwechikwata nekuda kwekufamba kwavo oscillatory, ichikwevera zvakasara mumusumbu nehelical motion (Vhidhiyo 4).Zvisinei, mumakumbo marefu, zvakaonekwa kuti mimwe misoro yembeu isingabhadharwi inonamira kumuviri uye chikamu chinogumira chetuft chinoshanda semavhane kuti chibatsire kufambisira mberi.
Paunenge uri mukuyerera zvishoma nezvishoma, masumbu embeu anofamba akaenzana kune mumwe nemumwe, zvisinei, anotanga kupindirana uye anonamatira kune zvose zvinenge zvichiri, kuitira kuti zvirege kushambidzwa nekuyerera kwezvino sezvo kuyerera kunowedzera.Misumbu inoumbwa kana tsama yemasero embeu inoswedera kune imwe, inotanga kufamba mu synchrony nekumonererana, uye ipapo inonamira pane chinhu chinonamira.Figure 1 ne2 inoratidza kuti urume hunosvikana sei, huchiumbana mhuno ichipotererana.
Vatsvakurudzi vakashandisa hydrostatic pressure kugadzira kuyerera kwemvura mune microchannel yekudzidza sperm rheology.A microchannel ine saizi ye200 µm × 20 µm (W × H) uye kureba kwe3.6 µm yakashandiswa.Shandisa microchannels pakati pemidziyo ine majekiseni akaiswa kumagumo.Kupenda kwekudya kwakashandiswa kuita kuti migero iwedzere kuoneka.
Sunga tambo dzekubatanidza uye zvishongedzo kumadziro.Vhidhiyo yakatorwa nechikamu chekusiyanisa maikorosikopu.Nemufananidzo wega wega, chikamu chekusiyanisa microscope uye mepu mifananidzo inoratidzwa.(A) Kubatana pakati pehova mbiri kunoramba kuyerera nekuda kwehelical motion (museve mutsvuku).(B) Kubatana pakati pechubhu bundle uye chiteshi chiteshi (miseve yakatsvuka), panguva imwechete iyo yakabatanidzwa kune mamwe masumbu maviri (yero miseve).(C) Masumbu embeu ari mugero remicrofluidic anotanga kubatana (miseve tsvuku), achigadzira mesh yemasumbu.(D) Kuumbwa kwe network ye sperm bundles.
Pakadonhedzwa donhwe reurume hwakanyungudutswa mumudziyo wemicrofluidic uye kuyerera kwakagadzirwa, danda rembeu raionekwa richienda kudivi rekuyerera.Misumbu inokwana zvakasimba pamadziro e microchannels, uye misoro yakasununguka muchikamu chekutanga chemasumbu inopindirana navo (vhidhiyo 5).Vanonamatira kune chero zvimedu zvakamira munzira yavo, zvakaita semarara, kuramba kukukurwa nemhepo.Nokufamba kwenguva, matinji aya anova mafilanti akareba achiteya imwe spermatozoa imwe chete uye mapfupi tufts (Vhidhiyo 6).Sezvo kuyerera kunotanga kuderera, mitsetse yakareba yembeu inotanga kuumba network ye sperm lines (Vhidhiyo 7; Mufananidzo 2).
Pakuyerera kwepamusoro (V > 33 µm/s), mafambiro eshinda eshinda anowedzerwa seyedzero yekubata urume hwakawanda hwemunhu huchigadzira masumbu zviri nani kuramba simba rekukukurwa rekuyerera. Pakuyerera kwepamusoro (V > 33 µm/s), mafambiro eshinda eshinda anowedzerwa seyedzero yekubata urume hwakawanda hwemunhu huchigadzira masumbu zviri nani kuramba simba rekukukurwa rekuyerera. При высокой скорости потока (V > 33 мкм/с) спиралевидные движения нитей усиливаются, оскольку они пытаются поймать множествоы оздоргий оздень х пучки, которые лучше противостоят дрейфующей силе потока. Pakuyerera kwepamusoro (V> 33 µm/s), kufamba kwehelical kwetambo kunowedzera sezvavanoedza kubata maspermatozoa akawanda achigadzira masumbu anonyatsokwanisa kurwisa simba rekudonha rekuyerera.在高流速(V > 33 µm/s) 时，螺纹的螺旋运动增加，以试图捕捉许多形成束的单个旺无子，漂移力。在 高 流速 (v> 33 µm/s) 时 , 的 螺旋 运动 增加 , 以漂移力。。。。。. При высоких скоростях потока (V > 33 мкм/с) спиральное движение нитей увеличивается в попытке захватить множество отделььное движение нитей увеличивается в попытке захватить множество отделььное движение нитей увеличивается чтобы лучше сопротивляться силам дрейфа потока. Pakuyerera kwepamusoro (V> 33 µm/s), kufamba kwehelical kwemafilaments kunowedzera mukuedza kubata akawanda maspermatozoa achigadzira mabundle kuti arambe zviri nani kukukurwa kwemasimba ekuyerera.Vakaedzawo kubatanidza ma microchannels kumasidewall.
Maspem bundles akaonekwa semasumbu emisoro yembeu uye inomonereka miswe uchishandisa light microscopy (LM).Maspem bundle ane akaunganidzwa akasiyana akaonekwawo semisoro yakamonyoroka uye ma flagellar aggregates, miswe yembeu yakawanda yakabatana, misoro yembeu yakanamatira kumuswe, uye misoro yembeu ine nuclei yakabhenda senyuchi dzakawanda dzakabatanidzwa.kuendesa electron microscopy (TEM).Scanning electron microscopy (SEM) yakaratidza kuti mabundle esperm aive akavharidzirwa akaunganidzwa emusoro wesperm uye masperm aggregates airatidza mambure akabatanidzwa emiswe yakamonerwa.
Iyo morphology uye ultrastructure yespermatozoa, kuumbwa kwespermatozoa masumbu akaongororwa achishandisa chiedza microscopy (hafu chikamu), scanning electron microscopy (SEM) uye transmission electron microscopy (TEM), sperm smears yakasvibiswa ne acridine orange uye yakaongororwa uchishandisa epifluorescence microscopy.
Smear smear staining ne acridine orange (Fig. 3B) yakaratidza kuti misoro yembeu yakanamatira pamwe chete uye yakafukidzwa nezvinhu zvepachivande, izvo zvakaita kuti pave nemhando huru (Fig. 3D).Masumbu embeu aive nembeu dzakaunganidzwa dzine network yemiswe yakasungirirwa (Fig. 4A-C).Mishonga yemasperm inoumbwa nemiswe yakawanda yespermatozoa yakanamatira pamwe chete (Fig. 4D).Zvakavanzika (Fig. 4E, F) yakavhara misoro yespermatozoa bundles.
Kuumbwa kwe spermatozoa bundle Kushandisa chikamu chekusiyanisa microscopy uye sperm smears yakasvibiswa ne acridine orange, yakaratidza kuti misoro yespermatozoa inonamatira pamwe chete.A(B) Photomicrograph yesperm smear yakasvibiswa neacridine orange inoratidza inonamatira misoro yembeu (miseve).Kubuda kunovhara musoro(s).Kukudza × 1000. (C) Kuvandudzwa kwedanda guru rinotakurwa nekuyerera mugero re microfluidic (uchishandisa high speed camera pa 950 fps).(D) Micrograph yesperm smear yakasvibiswa neacridine orange inoratidza mahombe mahombe (miseve).Kuwedzera: ×200.
Kuongorora electron micrograph yesperm beam uye smear yesperm yakasvibiswa neacridine orange.(A, B, D, E) idigital color scanning electron micrographs of spermatozoa, uye C and F mamicrographs e acridine orenji ane mavara esperm smears anoratidza kubatikana kweakawanda spermatozoa achiputira caudal web.(AC) Masperm aggregates anoratidzwa se network yemiswe yakasungirirwa (miseve).(D) Kunamira kwemaspermatozoa akati wandei (ane chinhu chinonamira, pink outline, museve) uchimonerera muswe.(E naF) Urume musoro aggregates (manongedzero) akafukidzwa anonamira zvinhu (anonongedzera).Iyo spermatozoa yakagadzira masumbu ane akati wandei mavhoti-sezvimiro (F).(C) × 400 uye (F) × 200 magnifications.
Tichishandisa hutachiwana erekitironi maikorosikopu, takaona kuti urume zvisumbu akanga akabatana miswe (Fig. 6A, C), misoro yakanamatira miswe (Fig. 6B), kana misoro yakanamatira miswe (Fig. 6D).Misoro yespermatozoa iri musumbu yakakombama, ichipa muchikamu chezvikamu zviviri zvenyukireya (Fig. 6D).Muchikamu chekucheka, spermatozoa yakanga ine musoro wakamonyaniswa nematunhu maviri enyukireya uye nzvimbo dzakawanda dzemureza (Fig. 5A).
Digital color electron micrograph inoratidza miswe yekubatanidza musperm bundle uye agglutinating material inobatanidza misoro yesperm.(A) Yakasungirirwa muswe wenhamba huru yespermatozoa.Tarisa uone kuti muswe unotaridzika sei mune zvese mufananidzo (museve) uye nyika (museve).(B) Musoro (museve) wesperm wakabatana nemuswe (museve).(C) Miswe yakawanda yembeu (miseve) yakasungirirwa.(D) Agglutination material (AS, blue) inobatanidza misoro mina yembeu (yepepuru).
Kuongorora ma electron microscopy kwaishandiswa kuona misoro yembeu mumasumbu embeu yakavharwa nezvakavharika kana membranes (Mufananidzo 6B), zvichiratidza kuti mabundle embeu aive akasungwa nekunze.The agglutinated zvinhu akanga concentrated mu urume musoro (jellyfish musoro-kufanana musangano; Fig. 5B) uye akawedzera disally, kupa inopenya yero chitarisiko pasi fluorescence microscopy kana yakasvibiswa acridine orenji (Fig. 6C).Chinhu ichi chinonyatso kuoneka pasi pemaikorosikopu uye chinoonekwa sechibhainda.Semi-yakaonda zvikamu (Fig. 5C) uye urume smears kusvibiswa ane acridine orenji akaratidza urume masvinga ane densely packed misoro uye curled miswe (Fig. 5D).
Akasiyana siyana mafotomicrographs anoratidza kuunganidzwa kwemisoro yesperm nemiswe yakapetwa uchishandisa nzira dzakasiyana.a(B) Digital color scanning electron micrograph inoratidza sumbu remisoro yembeu yakaita sejellyfish (miseve) inoita seyakavharwa.(C) Semi-tete chikamu chinoratidza aggregated urume misoro (miseve) uye akapetana miswe (miseve).(D) Micrograph yesperm smear yakasvibiswa neacridine orange inoratidza akaunganidzwa emusoro wembeu (miseve) uye yakamonera miswe (miseve).Ziva kuti chinhu chinonamira (S) chinovhara musoro wespermatozoon.(D) × 1000 kukura.
Kushandisa transmission electron microscopy (Fig. 7A), zvakaonekwawo kuti misoro yembeu yakamonyoroka uye nuclei yakanga ine spiral shape, sezvinosimbiswa nesperm smears yakasvibiswa ne acridine orange uye yakaongororwa uchishandisa fluorescence microscopy (Fig. 7B).
(A) Digital color transmission electron micrograph uye (B) Acridine orange stained sperm smear inoratidza misoro yakamonereka uye kubatanidzwa kwemusoro nemiswe yembeu (miseve).(B) × 1000 kukura.
Chinhu chinonakidza chawanikwa ndechekuti urume hwaSharkazi hunoungana kuita nharembozha filamentous bundle.Zvinhu zvezviputi izvi zvinotibvumira kuti tinzwisise basa ravo rinobvira mukutorwa uye kuchengetedza spermatozoa muSST.
Mushure mekusangana, urume hunopinda mubeche roita sarudzo dzakasimba, zvichiita kuti pave nenhamba shoma yembeu inopinda muSST15,16.Kusvika iye zvino, maitiro ekuti urume hunopinda nekubuda muSST hazvina kujeka.Muhuku, spermatozoa inochengetwa muSST kwenguva yakareba yemavhiki maviri kusvika gumi, zvichienderana nemhando6.Kukakavadzana kunoramba kuri pamusoro pemamiriro eurume panguva yekuchengetedza muSST.Vari kufamba here kana kuti vakazorora?Mune mamwe mazwi, maseru eurume anochengetedza sei chinzvimbo chavo muSST kwenguva yakareba kudaro?
Forman4 vakakurudzira kuti kugara kweSST uye kubuda kunogona kutsanangurwa maererano nekufamba kwembeu.Vanyori vanofungidzira kuti urume hunochengetedza nzvimbo yavo nekushambira kuyerera kwemvura inogadzirwa neSST epithelium uye kuti urume hunoburitswa kubva kuSST kana velocity yavo ichiwira pasi peiyo painotangira kudzokera kumashure nekuda kwekushaya simba.Zaniboni5 yakasimbisa kuvapo kweaquaporins 2, 3, uye 9 muchikamu cheapical cheSST epithelial cells, iyo inogona kutsigira nenzira isina kunanga yeForeman's sperm storage model.Muchidzidzo chezvino, takaona kuti inenge hafu yeSharkashi's spermatozoa inoratidza rheology yakanaka mumvura inoyerera, uye kuti masumbu eurume agglutinated anowedzera nhamba yespermatozoa inoratidza rheology yakanaka, kunyange zvazvo agglutination inononoka.Mafambiro anoita masero embeu achikwira nechubhu reshiri kusvika painosangana hazvinyatsonzwisiswi.Mumhuka dzinoyamwisa, follicular fluid chemoattracts spermatozoa.Zvisinei, mishonga yemishonga inotendwa kuti inonangisa spermatozoa kusvika kure kure7.Nokudaro, dzimwe nzira dzine mutoro wekutakura urume.Kugona kwembeu kutenderera uye kuyerera ichipikisana ne fallopian chubhu fluid inoburitswa mushure mekusangana kwanzi ndicho chinhu chikuru mukunangana nembeu mumakonzo.Parker 17 akakurudzira kuti spermatozoa iyambuke maoviducts nekushambira ichirwisana neciliary current mushiri nezvinokambaira.Kunyangwe zvisina kumboratidzwa mushiri, Adolphi18 ndiye akatanga kuona kuti urume hweavian hunopa mhedzisiro kana mutete wemvura pakati pechivharo nechisiraidhi uchigadzirwa nemucheka webepa resefa.Rheology.Hino naYanagimachi [19] vakaisa mbeva ovary-tubal-uterine complex mu perfusion ring ndokubaya 1 µl yeinki muisthmus kuti vaone kuyerera kwemvura mu fallopian chubhu.Vakaona kufamba kwakanyanyisa kwekumanikana uye kuzorora mu fallopian chubhu, umo mabhora einki ese aifamba akananga ku ampulla ye fallopian chubhu.Vanyori vanosimbisa kukosha kwekuyerera kwe tubal fluid kubva pasi kuenda kupamusoro fallopian chubhu kuitira kusimudza urume nekusangana.Brillard20 yakashuma kuti muhuku uye turkeys, spermatozoa inotama nekufamba-famba kubva pamusuwo wemukadzi, kwainochengeterwa, ichienda kune utero-vaginal junction, kwainochengetwa.Nekudaro, kufamba uku hakudiwi pakati peiyo uterovaginal junction uye infundibulum nekuti iyo spermatozoa inotakurwa nekungofamba-famba.Kuziva izvi zvinokurudzira zvakapfuura uye mhedzisiro yakawanikwa muchidzidzo chezvino, zvinogona kufungidzirwa kuti kugona kwespermatozoa kukwira kumusoro (rheology) ndeimwe yezvivakwa izvo kusarudzwa kwakavakirwa.Izvi zvinogadzirisa nzira ye spermatozoa kuburikidza nemukati uye kupinda kwavo muCCT yekuchengetedza.Sezvakataurwa neForman4, izvi zvinogona zvakare kufambisa nzira yembeu ichipinda muSST nenzvimbo yayo kwenguva yakati rebei yobva yabuda kana kumhanya kwadzo kunotanga kudzikira.
Kune rimwe divi, Matsuzaki naSasanami 21 vakakurudzira kuti avian spermatozoa iite shanduko yekuchinja kubva kudormancy kuenda kumotility mumaturakiti echirume neechikadzi.Inhibition yevagari vembeu motility muSST yakatsanangurwa kutsanangura iyo yakareba yekuchengetedza nguva yembeu uyezve rejuvenation mushure mekusiya SST.Pasi pe hypoxic mamiriro, Matuzaki et al.1 yakashuma kugadzirwa kwakanyanya uye kuburitswa kwelactate muSST, izvo zvinogona kutungamira mukudzivirirwa kwekugara kwembeu motility.Muchiitiko ichi, kukosha kwesperm rheology kunoratidzwa mukusarudzwa uye kunwa kwespermatozoa, uye kwete mukuchengetedza kwavo.
Iyo sperm agglutination pattern inoonekwa setsananguro inonzwisisika yenguva refu yekuchengetedza urume muSST, sezvo iyi iri nzira yakajairika yekuchengeta urume muhuku2,22,23.Bakst et al.2 yakacherechedza kuti spermatozoa yakawanda yakanamatira kune imwe neimwe, ichigadzira fascicular aggregates, uye imwe spermatozoa yaisawanzowanikwa muzvihuta CCM.Kune rumwe rutivi, Wen et al.24 yakacherekedza yakawanda yakapararira spermatozoa uye mashoma maspermatozoa tufts muSST lumen muhuku.Kubva pane zvakaonekwa izvi, zvinogona kufungidzirwa kuti propensity ye sperm agglutination inosiyana pakati peshiri uye pakati pe spermatozoa mune imwechete ejaculate.Mukuwedzera, Van Krey et al.9 yakaratidza kuti kupatsanurwa zvisina tsarukano kweagglutinated spermatozoa kunokonzera kupinda zvishoma nezvishoma kwespermatozoa mu lumen ye fallopian chubhu.Maererano nefungidziro iyi, spermatozoa ine yakaderera agglutination kugona inofanira kudzingwa kubva kuSST kutanga.Muchirevo chechinyorwa chino, kugona kwespermatozoa kuagglutinate kungave chinhu chinokanganisa mhedzisiro yemakwikwi eurume mushiri dzine tsvina.Pamusoro pazvo, kana urume hwakanyungudutswa huchiparadzana kwenguva refu, kubereka kunochengetedzwa.
Kunyange zvazvo spermatozoa aggregation uye aggregation mumasumbu zvakaonekwa mune dzakawanda zvidzidzo2,22,24, hazvina kutsanangurwa zvakadzama nekuda kwekuoma kwekucherechedza kwavo kinematic mukati meSST.Kuedza kunoverengeka kwakaitwa kudzidza sperm agglutination in vitro.Kuunganidza kwakawanda asi kwenguva pfupi kwakaonekwa pakabviswa waya yakatetepa kubva padonhwe rembeu rakarembera.Izvi zvinotungamira kune chokwadi chekuti bubble rakareba rinobuda kubva pakudonha, richitevedzera seminal gland.Nekuda kwekugumira kwe3D uye nguva pfupi yekudonha yekudonha, iyo block yese yakakurumidza kuwira mukusagadzikana9.Muchidzidzo chazvino, tichishandisa huku Sharkashi uye microfluidic chips, takakwanisa kutsanangura kuti matuft aya anoumbwa sei uye kuti anofamba sei.Masumbu esperm akaumbwa pakarepo mushure mekuunganidzwa kwembeu uye akawanikwa achifamba mumhepo, achiratidza rheology yakanaka kana iripo mukuyerera.Uyezve, kana macroscopically ichitariswa, sperm bundles akacherechedzwa kuti awedzere mutsara wemotility uchienzaniswa neiyo yega spermatozoa.Izvi zvinoratidza kuti kusanganiswa kwembeu kunogona kuitika SST isati yapinda uye kuti kugadzirwa kwembeu hakugumiri kunzvimbo diki nekuda kwekunetsekana sezvakambotaurwa (Tingari neLake12).Panguva yekugadzira tuft, spermatozoa inotuhwina mu synchrony kusvika yaita makutano, ipapo miswe yadzo inomonerana uye musoro wespermatozoon unoramba wakasununguka, asi muswe nechikamu chepasi che spermatozoon zvinonamatira pamwe nechinhu chinonamira.Nokudaro, musoro wakasununguka wemutsara une mutoro wekufambisa, uchikwevera mamwe mazamu.Kuongorora ma electron microscopy emasumbu esperm airatidza misoro yesperm yakavharirwa nezvinhu zvakawanda zvinonamira, zvichiratidza kuti misoro yembeu yakasungirirwa mumasumbu ekuzorora, izvo zvinogona kunge zvakaitika mushure mekusvika panzvimbo yekuchengetera (SST).
Kana sperm smear yakasvibiswa ne acridine orange, extracellular adhesive material yakatenderedza sperm cells inogona kuonekwa pasi pe fluorescent microscope.Ichi chinhu chinobvumira masumbu embeu kuti anamate uye anamira kune chero nzvimbo dzakatenderedza kana zvidimbu kuti zvisakukurwa nekuyerera kwakatenderedza.Nokudaro, maonero edu anoratidza basa re spermatozoa adhesion mumhando ye mobile bundles.Kugona kwavo kushambira vachipesana nezvino uye kunamatira kunzvimbo dziri pedyo kunobvumira urume kugara kwenguva refu muSST.
Rothschild25 yakashandisa hemocytometry kamera kudzidza kuyerera kwembeu yebovi mudonhwe rekumiswa, ichitora mafotomicrographs kuburikidza nekamera ine ese ari maviri akatwasuka uye akachinjika optical axis yemaikorosikopu.Zvigumisiro zvakaratidza kuti spermatozoa yakakwezvwa pamusoro pekamuri.Vanyori vanoratidza kuti panogona kunge paine kupindirana kwehydrodynamic pakati pesperm nepamusoro.Tichifunga izvi, pamwe nekugona kweSharkashi chick semen kugadzira tuft tuft, zvinogona kuwedzera mukana wekuti mbeu inonamatira kumadziro eSST uye kuchengetwa kwenguva yakareba.
Bccetti naAfzeliu26 vakashuma kuti sperm glycocalyx inodiwa kuti gamete izivikanwe uye agglutination.Forman10 yakacherechedza kuti hydrolysis ye-α-glycosidic bonds mu glycoprotein-glycolipid coatings nekurapa mbeu yeavian neuraminidase yakakonzera kuderedza kubereka pasina kukanganisa urume kufamba.Vanyori vanoratidza kuti mhedzisiro yeneuraminidase pa glycocalyx inokanganisa kutorwa kwembeu pautero-vaginal junction, nekudaro ichideredza kubereka.Zvavanoona hazvigone kufuratira mukana wekuti kurapwa neuraminidase kunogona kuderedza urume uye kuzivikanwa kweoocyte.Forman naEngel10 vakaona kuti mbereko yakadzikira apo huku dzaipinzwa mutsinga nembeu yakarapwa neuraminidase.Zvakadaro, IVF ine neuraminidase yakarapwa urume haina kukanganisa kubereka kana zvichienzaniswa nehuku dzekudzora.Vanyori vakagumisa kuti kuchinja kweglycoprotein-glycolipid coating kutenderedza sperm membrane kwakaderedza kugona kwembeu kuti isanganise nekukanganisa kutorwa kwembeu pautero-vaginal junction, izvo zvakazowedzera kurasikirwa kwembeu nekuda kwekumhanya kweutero-vaginal junction, asi kwete kukanganisa urume uye kucherechedzwa kwezai.
Mumaturkeys Bakst naBauchan 11 vakawana mavhesi madiki uye zvidimbu zvemembrane mulumen yeSST uye vakaona kuti mamwe emagranules aya akange asangana nesperm membrane.Vanyori vanoratidza kuti hukama uhwu hunogona kubatsira pakuchengetedza kwenguva refu kwespermatozoa muSST.Zvisinei, vatsvakurudzi havana kutsanangura kwakabva zvimedu izvi, zvingave zvakavanzwa neCCT epithelial masero, anogadzirwa uye akavanzwa nehurongwa hwekubereka kwemurume, kana kugadzirwa nembeu pachayo.Zvakare, zvimedu izvi zvinokonzeresa agglutination.Grützner et al27 vakashuma kuti masero epididymal epithelial anogadzira uye anobudisa puroteni chaiyo inodiwa kuti pave ne-single-pore seminal tracts.Vanyori vanotaurawo kuti kupararira kwemasumbu aya kunoenderana nekudyidzana kwe epididymal proteins.Nixon et al28 vakawana kuti adnexa inobudisa puroteni, iyo acidic cysteine-rich osteonectin;SPARC inobatanidzwa mukugadzirwa kwesperm tufts mune pfupi-beaked echidnas uye platypus.Kupararira kwematanda aya kunoenderana nekurasikirwa kweprotein iyi.
Muchidzidzo chemazuva ano, kuongorora kwekugadzirwa kwemagetsi uchishandisa electron microscopy kwakaratidza kuti spermatozoa yakanamatira kune yakawanda yakawanda yezvinhu.Zvinhu izvi zvinofungidzirwa kuti ndizvo zvinoita kuti agglutination isanganise pakati nepakati pemisoro inonamatira, asi pakadzika pasi munzvimbo yemuswe.Isu tinofungidzira kuti chinhu ichi chinokonzeresa chinoburitswa kubva kumurume wekubereka (epididymis kana vas deferens) pamwe nembeu, sezvo isu kazhinji tinoona urume huchiparadzana nelymph uye seminal plasma panguva yekubuda.Izvo zvave zvichishumwa kuti sezvo avian spermatozoa inopfuura nepakati epididymis uye vas deferens, vanopinda maturation-anoenderana nekuchinja anotsigira kugona kwavo kusunga mapuroteni uye kuwana plasma lemma-inosanganiswa glycoproteins.Kuenderera mberi kwemapuroteni aya pane anogara masperm membranes muSST anoratidza kuti mapuroteni aya anogona kukanganisa kuwanikwa kwesperm membrane kugadzikana 30 uye kuona kubereka kwavo 31.Ahammad et al32 vakashuma kuti spermatozoa yakawanikwa kubva kumativi akasiyana-siyana ehutano hwemurume (kubva kuma testes kusvika kune distal vas deferens) yakaratidza kuwedzera kunofambira mberi kwehutano pasi pemamiriro ekuchengetedza mvura, pasinei nekupisa kwekuchengetedza, uye hupenyu hwehuku hunowedzerawo mumachubhu e fallopian mushure mekuiswa kwechigadzirwa.
Sharkashi huku sperm tufts ane hunhu hwakasiyana uye mabasa pane mamwe marudzi akadai se echidnas, platypus, mbeva dzehuni, mhembwe, uye guinea pigs.Muhuku dze sharkasi, kuumbwa kwe spermatozoa bundles kwakaderedza kushambira kwavo kuenzaniswa nespermatozoa imwe chete.Nekudaro, masumbu aya akawedzera chikamu che rheologically positive spermatozoa uye akawedzera kugona kwespermatozoa kuzvidzikamisa munzvimbo ine simba.Saka, mhedzisiro yedu inosimbisa zano rekare rekuti sperm agglutination muSST inosanganiswa nekugara kwenguva refu urume kuchengetedza.Isu tinofungidzirawo kuti humiro hweurume hwekuita tufts hunogona kudzora mwero wekurasikirwa kwembeu muSST, izvo zvinogona kushandura mhedzisiro yemakwikwi embeu.Zvinoenderana nefungidziro iyi, spermatozoa ine yakaderera agglutination kugona kuburitsa SST kutanga, nepo spermatozoa ine yakakwira agglutination kugona inoburitsa vazhinji vevana.Kuumbwa kwe-single-pore sperm bundles kunobatsira uye kunokanganisa chiyero chemubereki-mwana, asi inoshandisa imwe nzira.Mune echidnas uye platypus, spermatozoa inorongedzerwa yakafanana kune imwe neimwe kuti iwedzere kukurumidza kumberi kwedanda.Masumbu eechidna anofamba kanenge katatu nekukurumidza kupfuura imwe spermatozoa.Zvinotendwa kuti kuumbwa kwembeu dzakadai muechidnas ishanduko yekushanduka-shanduka kuchengetedza kutonga, sezvo vakadzi vanoita hupombwe uye kazhinji vanosangana nevarume vakati wandei.Nokudaro, spermatozoa kubva kune zvakasiyana-siyana ejaculates inokwikwidza zvakasimba pakubatanidzwa kwezai.
Agglutinated spermatozoa yehuku dze sharkasi zviri nyore kuona uchishandisa chikamu chekusiyanisa microscopy, iyo inoonekwa seyakanakira nekuti inobvumira kudzidza nyore kwemaitiro espermatozoa in vitro.Iyo nzira iyo iyo spermatozoa inogadzirwa inosimudzira kuberekana muhuku dze sharkasi zvakare yakasiyana neiyo inoonekwa mune dzimwe placenta inoyamwisa inomiririra kubatana kwembeu maitiro sehuni mbeva, uko imwe spermatozoa inosvika mazai, ichibatsira vamwe vanhu vane hukama kusvika nekukuvadza mazai avo.kuti uzviratidze iwe pachako.kuzvibata.Self-fertilization 34. Mumwe muenzaniso wehutano hwekubatana mu spermatozoa wakawanikwa mumakonzo emhembwe, apo spermatozoa yakakwanisa kuziva uye kusanganisa ne spermatozoa yakawanda yakabatana uye inoumba mapoka ekubatana kuti awedzere kukurumidza kwavo kana ichienzaniswa nespermatozoa35 isingabatanidzi.
Zvigumisiro zvakawanikwa muchidzidzo ichi hazvipikisani nedzidziso yeFoman yekuchengetedza kwenguva refu yespermatozoa muSWS.Vatsvakurudzi vanotaura kuti masero embeu anoramba achifamba mukuyerera kwemasero epithelial akavhara SST kwenguva yakawedzerwa, uye mushure meimwe nguva yenguva, zvitoro zvesimba zvemasero eurume zvinopera, zvichikonzera kuderera kwekukurumidza, izvo zvinobvumira kudzingwa kwediki mamorekuru zvinhu.simba re spermatozoa nekuyerera kwemvura kubva mu lumen ye SST Mugoti we fallopian tube.Muchidzidzo chezvino, takaona kuti hafu yembeu imwe chete yakaratidza kukwanisa kushambira pamusoro pemvura inoyerera, uye kunamatira kwavo mumusumbu kwakawedzera kukwanisa kwavo kuratidza rheology yakanaka.Uyezve, data redu rinoenderana neaMatsuzaki et al.1 uyo akataura kuti yakawedzera lactate secretion muSST inogona kutadzisa mugari wembeu motility.Nekudaro, mhedzisiro yedu inotsanangura kuumbwa kwesperm motile ligaments uye maitiro avo erheological pamberi penzvimbo ine simba mukati meiyo microchannel mukuyedza kujekesa maitiro avo muSST.Tsvagiridzo yeramangwana inogona kutarisa pakuona kuumbwa kwemakemikari uye kwakabva agglutinating agent, izvo pasina mubvunzo zvichabatsira vaongorori kugadzira nzira nyowani dzekuchengeta urume hwemvura uye kuwedzera nguva yekubereka.
Gumi neshanu 30-vhiki-yekuzvarwa isina-mutsipa murume sharkasi (homozygous dominant; Na Na) vakasarudzwa sevanopa urume muchidzidzo.Shiri idzi dzakachengetwa paResearch Poultry Farm yeFaculty of Agriculture, Ashit University, Ashit Governorate, Egypt.Shiri dzakaiswa mumakeji ega (30 x 40 x 40 cm), pasi pechirongwa chechiedza (16 maawa echiedza uye maawa 8 erima) uye akadyisa kudya kuine 160 g yemapuroteni asina kuchena, 2800 kcal yesimba rinogadziriswa, 35 g ye calcium imwe neimwe.5 magiramu ephosphorus inowanikwa pakirogiramu yezvokudya.
Zvinoenderana nedata 36, ​​37, urume hwakaunganidzwa kubva kuvarume neabdominal massage.Huwandu hwe45 semen samples dzakaunganidzwa kubva kuvarume gumi nevashanu mumazuva matatu.Semen (n = 15 / zuva) yakakurumidza kuderedzwa 1: 1 (v: v) neBelsville Poultry Semen Diluent, ine potassium diphosphate (1.27 g), monosodium glutamate monohydrate (0.867 g), fructose (0.5 d) anhydrous sodium.acetate (0.43 g), tris(hydroxymethyl)aminomethane (0.195 g), potassium citrate monohydrate (0.064 g), potassium monophosphate (0.065 g), magnesium chloride (0.034 g) uye H2O (100 ml), pH = 7, 5, osmolarity 3/33 m33 m.Semen yakanyungudutswa yakatanga kuongororwa pasi pemaikorosikopu yakareruka kuti ive nechokwadi chekunaka kwembeu (unyoro) uye yozochengetwa mubhavhu remvura pa37 ° C kusvika yashandiswa mukati mehafu yeawa mushure mekuunganidzwa.
Iyo kinematics uye rheology ye spermatozoa inotsanangurwa uchishandisa system ye microfluidic zvishandiso.Semen samples yakawedzera kuderedzwa kusvika 1:40 muBeltsville Avian Semen Diluent, yakarongedzerwa muine microfluidic mudziyo (ona pazasi), uye kinetic parameters yakatemwa uchishandisa Computerized Semen Analysis (CASA) system yakambogadzirwa kuti microfluidics hunhu.pamusoro pekufamba kwespermatozoa mumvura yemvura (Dhipatimendi reMechanical Engineering, Faculty of Engineering, Assiut University, Egypt).Plugin inogona kutorwa pa: http://www.assiutmicrofluidics.com/research/casa39.Curve velocity (VCL, μm/s), linear velocity (VSL, μm/s) uye avhareji trajectory velocity (VAP, μm/s) akayerwa.Mavhidhiyo espermatozoa akatorwa pachishandiswa inverted Optika XDS-3 phase musiyano maikorosikopu (ine 40x chinangwa) yakabatana neTucson ISH1000 kamera pa30 fps kwe3 s.Shandisa iyo CASA software kudzidza zvinosvika nzvimbo nhatu uye mazana mashanu eurume trajectories pamuenzaniso.Vhidhiyo yakarekodhwa yakagadziriswa pachishandiswa CASA yekumba.Tsanangudzo yemotility muCASA plug-in inobva pakushambira kwechirume chichienzaniswa nekuyerera kwechirume, uye haisanganisi mamwe maitiro akadai sekufamba-kurutivi-kurutivi, sezvo izvi zvakawanikwa zvichinyanya kuvimbika mukuyerera kwemvura.Rheological motion inotsanangurwa sekufamba kwesperm cells ichipokana negwara rekuyerera kwemvura.Spermatozoa ine rheological properties yakakamurwa nenhamba ye motile spermatozoa;spermatozoa yakanga yazorora uye convectively inofamba spermatozoa yakabviswa kubva pakuverenga.
Makemikari ese akashandiswa akawanikwa kubva kuElgomhoria Pharmaceuticals (Cairo, Egypt) kunze kwekunge zvataurwa neimwe nzira.Chigadzirwa chacho chakagadzirwa sezvakatsanangurwa naEl-sherry et al.40 pamwe nezvimwe zvakagadziridzwa.Zvinhu zvakashandiswa kugadzira ma microchannels zvaisanganisira magirazi egirazi (Howard Glass, Worcester, MA), SU-8-25 negative resist (MicroChem, Newton, CA), diacetone alcohol (Sigma Aldrich, Steinheim, Germany), uye polyacetone.-184, Dow Corning, Midland, Michigan).Microchannels inogadzirwa uchishandisa soft lithography.Kutanga, yakajeka inodzivirira kumeso mask ine yaidiwa microchannel dhizaini yakadhindwa pane yakakwirira resolution printer (Prismatic, Cairo, Egypt uye Pacific Arts uye Dhizaini, Markham, ON).Iwo masters akagadzirwa achishandisa magirazi egirazi se substrates.Mahwendefa acho akacheneswa muacetone, isopropanol uye mvura yakasvibiswa uye akazoputirwa ne20 µm layer yeSU8-25 ne spin coating (3000 rpm, 1 min).Iwo SU-8 akaturikidzana akaomeswa zvinyoro nyoro (65 ° C, 2 min uye 95 ° C, 10 min) uye akaiswa kune UV mwaranzi kwe50 s.Post-exposure bika pa 65 ° C uye 95 ° C kwe 1 min uye 4 min kuti crosslink yakazaruka SU-8 layers, inoteverwa nekuvandudzwa mu diacetone doro kwe 6.5 min.Zvakaoma kubika waffles (200 ° C kwemaminetsi gumi nemashanu) kuti uwedzere kusimbisa iyo SU-8 layer.
PDMS yakagadzirwa nekusanganisa monoma uye hardener muhuremu hwe10: 1, ndokuzobviswa muvacuum desiccator ndokudururirwa paSU-8 huru furemu.Iyo PDMS yakaporeswa muchoto (120 ° C, 30 min), ipapo migero yakatemwa, yakaparadzaniswa kubva kuna tenzi, uye perforated kubvumira machubhu kuti abatanidzwe pakupinza uye kubuda kweiyo microchannel.Pakupedzisira, PDMS microchannels dzakabatanidzwa zvachose kune maikorosikopu masiraidhi vachishandisa inotakurika corona processor (Electro-Technic Zvigadzirwa, Chicago, IL) sezvakatsanangurwa kumwe kunhu.Iyo microchannel inoshandiswa muchidzidzo ichi inoyera 200 µm × 20 µm (W × H) uye yakareba 3.6 cm.
Mvura inoyerera inokonzerwa ne hydrostatic pressure mukati me microchannel inowanikwa nekuchengetedza chiyero chemvura mumudziyo wekupinza pamusoro pekusiyana kwehurefu Δh39 mumudziyo wekubuda (Fig. 1).
apo f ndiyo coefficient of friction, inotsanangurwa se f = C / Re ye laminar inoyerera mumutsara une rectangular, apo C inogara ichienderana nechikamu chechikamu chechiteshi, L ndiyo urefu hwe microchannel, Vav ivhareji yevelocity mukati me microchannel, Dh ndiyo hydraulic diameter ye chiteshi, g - kukurumidza kwegiravhiti.Uchishandisa iyi equation, avhareji chiteshi velocity inogona kuverengerwa uchishandisa inotevera equation: