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Kubereka kweshiri kunoenderana nekukwanisa kwadzo kuchengeta mbeu yakakwana kwenguva yakareba mu tubules dzekuchengetera mbeu (SST). Nzira chaiyo iyo mbeu yemurume inopinda, inogara, uye ichibva muSST ichiri kukakavadzana. Hurume hwehuku dzesharkasi hwakaratidza katsika kekusangana, zvichigadzira mabundles anofambika ane masero akawanda. Nekuda kwekuoma kwekuona kufamba uye maitiro ehurume muchubhu yefallopian isina chinhu, takashandisa mudziyo we microfluidic une microchannel cross-section yakafanana neye spermatozoa kudzidza spermatozoa agglutination uye kufamba. Chidzidzo ichi chinokurukura kuti mabundles ehurume anoumbwa sei, kuti anofamba sei, uye basa rawo rinogona kuitwa mukuwedzera kugara kwehurume muSST. Takaongorora kumhanya kwehurume uye maitiro erheological apo kuyerera kwemvura kwakagadzirwa mukati me microfluidic channel ne hydrostatic pressure (flow rate = 33 µm/s). Hurume hunowanzo shambira huchipesana ne current (positive rheology) uye kumhanya kwehurume hurume kunoderedzwa zvakanyanya kana tichienzanisa nehurume humwechete. Maponde embeu akaonekwa achifamba-famba uye achiwedzera kureba nekukora sezvo mbeu imwe chete ichiunganidzwa. Mapaketi embeu akaonekwa achiswedera uye achinamatira pamativi enzira dze microfluidic kudzivirira kutsvairwa nekumhanya kwekuyerera kwemvura > 33 µm/s. Mapaketi embeu akaonekwa achiswedera uye achinamatira pamativi enzira dze microfluidic kudzivirira kutsvairwa nekumhanya kwekuyerera kwemvura > 33 µm/s. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрофлюидных каналов, чтобы избежают смета жидкости> 33 мкм / p. Mapaketi embeu akaonekwa achiswedera pedyo nekunamatira pamadziro emativi emigero ye microfluidic kudzivirira kukweverwa nekuyerera kwemvura >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 сместо приближаются и прилипают к боковым стенкам микрожидкостного канала, чтобы избежать смести смести 33 мкм/с. Mapaketi embeu akaonekwa achiswedera pedyo nekunamatira pamadziro erutivi rwemugero we microfluidic kudzivirira kukweverwa nemvura inoyerera pa >33 µm/s.Kuongororwa nekushandiswa kwemaelektroni emagetsi kwakaratidza kuti mabundle embeu aitsigirwa nezvinhu zvakawanda zvakakora. Ruzivo rwakawanikwa runoratidza kufamba kwakasiyana kwembeu yehuku yeSharkazi, pamwe nekukwanisa kwembeu yehuku kubatana nekugadzira mabundle ekufamba, izvo zvinobatsira kunzwisisa zviri nani kuchengetwa kwenguva refu kwembeu yehuku muSMT.
Kuti vanhu nemhuka zhinji vawane kusangana kwembeu, mbeu nemazai zvinofanira kusvika panzvimbo yekusangana kwembeu panguva chaiyo. Nokudaro, kusangana kwembeu kunofanira kuitika nguva isati yasvika kana kuti panguva yekusangana kwembeu. Kune rumwe rutivi, dzimwe mhuka dzinoyamwisa, dzakadai sembwa, pamwe chete nedzimwe mhuka dzisiri mhuka dzinoyamwisa, dzakadai sezvipembenene, hove, zvinokambaira, neshiri, dzinochengeta mbeu munhengo dzadzo dzekubereka kwenguva yakareba kusvika mazai agadzirira kusangana (kusangana kwembeu 1). Shiri dzinokwanisa kuchengetedza simba rembeu dzinokwanisa kusangana nemazai kwemavhiki maviri kusvika gumi.
Ichi chinhu chakasiyana chinosiyanisa shiri nedzimwe mhuka, sezvo zvichiita kuti pave nemukana mukuru wekusangana kwembeu mushure mekuberekeswa kamwe chete kwemavhiki akati wandei pasina kusangana uye ovulation panguva imwe chete. Nhengo huru yekuchengetera mbeu, inonzi sperm storage tubule (SST), iri mumakomba emukati memucosal panosangana uterovaginal. Kusvika pari zvino, nzira dzinoshandiswa nembeu kupinda, kugara, uye kubuda mubhengi rembeu hadzina kunzwisiswa zvizere. Zvichibva pane zvidzidzo zvekare, fungidziro dzakawanda dzakaiswa, asi hapana kana imwe yadzo yakasimbiswa.
Forman4 akafungidzira kuti mbeu dzemurume dzinoramba dzichigara muSST cavity kuburikidza nekufamba-famba kunoenderera mberi nekufamba kwemvura kuburikidza nemapuroteni ari paSST epithelial cells (rheology). ATP inodzikira nekuda kwekushanda kweflagellar kunodiwa kuchengetedza mbeu dzemurume muSST lumen uye kufamba-famba kunozopedzisira kwaderera kusvika mbeu dzemurume dzabuda mubhengi rembeu nekuyerera kwemvura uye kutanga rwendo rutsva pasi pechubhu inokwira yefallopian kuti iberekese mbeu. Zai (Forman4). Iyi modhi yekuchengetwa kwembeu inotsigirwa nekuonekwa ne immunocytochemistry ye aquaporins 2, 3 na9 dziri mumasero eSST epithelial. Kusvika pari zvino, zvidzidzo pamusoro pehuku rheology uye basa rayo mukuchengetedza SST, kusarudza mbeu dzemurume, uye kukwikwidzana kwembeu hazvipo. Muhuku, mbeu dzemurume dzinopinda muhuku mushure mekusangana kwechisikigo, asi anopfuura 80% yembeu dzinoburitswa muhuku nguva pfupi mushure mekusangana. Izvi zvinoratidza kuti hunhu hwemurume ndiyo nzvimbo huru yekusarudzwa kwembeu mushiri. Pamusoro pezvo, zvakataurwa kuti pasi pe1% yembeu dzinoberekeswa muchibereko dzinoguma dzave muSST2. Mukuisa nhiyo dzevana muchibereko, huwandu hwembeu dzinosvika kuSST hunowanzowedzera maawa makumi maviri nemana mushure mekuisa nhiyo. Kusvika pari zvino, mashandiro ekusarudza mbeu panguva iyi haana kujeka, uye kufamba kwembeu kunogona kuita basa rakakosha mukutora mbeu dzeSST. Nekuda kwemadziro akakora uye asina kujeka emachubhu efallopian, zvakaoma kutarisa zvakananga kufamba kwembeu mumachubhu efallopian eshiri. Nokudaro, hatina ruzivo rwekuti mbeu dzinoshanduka sei kuenda kuSST mushure mekuisa nhiyo.
Rheology yave kuzivikanwa munguva pfupi yapfuura sechinhu chakakosha chinodzora kutakurwa kwembeu munhengo dzesikarudzi dzemhuka dzinoyamwisa. Zvichienderana nekukwanisa kwembeu kufamba-famba kunofamba-famba, Zaferani nevamwe vakashandisa corra microfluidic system kuti vabvise mbeu dzinofamba-famba kubva muzvimedu zvembeu zvakavharirwa. Rudzi urwu rwekuronga mbeu rwakakosha pakurapa kusabereka kwevana uye kutsvagurudza kwekiriniki, uye rwunofarirwa pane nzira dzechinyakare dzinotora nguva uye dzinoda kushanda nesimba uye dzinogona kukanganisa chimiro chembeu uye kusimba kwechimiro. Zvisinei, kusvika pari zvino, hapana zvidzidzo zvakaitwa pamusoro pemhedzisiro yekubuda kwembeu kubva munhengo dzesikarudzi dzehuku pakufamba kwembeu.
Pasinei nemaitiro anoita kuti mbeu yemurume irambe yakachengetwa muSST, vaongorori vazhinji vakaona kuti mbeu yemurume inogara muhuku dzinobatana muSST yehuku 9, 10, quail 2, uye turkeys 11 kuti dzigadzire nhengo dzehurume dzakasanganiswa. Vanyori vanoti pane hukama pakati pekubatana uku nekuchengetwa kwenguva refu kwembeu dzemurume muSST.
Tingari naLake12 vakataura nezvehukama hwakasimba pakati pembeu dziri mugland rinogamuchira urume rehuku uye vakabvunza kana mbeu dzembeu dzeshiri dzichibatanidzwa nenzira imwechete nembeu dzemhuka dzinoyamwisa. Vanotenda kuti kubatana kwakadzika pakati pembeu dziri muvas deferens kunogona kunge kuri nekuda kwekushushikana kunokonzerwa nekuvapo kwembeu dzakawanda munzvimbo diki.
Pakuongorora maitiro embeu dzemudumbu pamasiraidhi matsva egirazi, zviratidzo zvenguva pfupi zvekubatana kwembeu dzemudumbu zvinogona kuonekwa, kunyanya pamucheto wemadonhwe embeu dzemudumbu. Zvisinei, kubatana kwembeu dzemudumbu kwaiwanzokanganiswa nekufamba-famba kwakabatana nekufamba-famba kwenguva refu, izvo zvinotsanangura hunhu hwechiitiko ichi. Vaongorori vakaonawo kuti apo musanganiswa wakawedzerwa kumbeu dzemudumbu, masero akareba "akaita seshinda" akaonekwa.
Kuedza kwekutanga kutevedzera spermatozoon kwakaitwa nekubvisa waya yakatetepa kubva padonhwe rakarembera, izvo zvakaita kuti vesicle yakareba yakaita sesperm ibude kubva padonhwe rembeu. spermatozoa yakabva yangomira yakafanana mukati me vesicle, asi chinhu chose chakakurumidza kunyangarika nekuda kwemiganhu ye3D. Saka, kuti udzidze spermatozoa agglutination, zvakakosha kuona kufamba uye maitiro espermatozoa zvakananga mumatubule ekuchengetera sperm akaparadzaniswa, izvo zvakaoma kuita. Saka, zvakakosha kugadzira chishandiso chinotevedzera spermatozoa kutsigira zvidzidzo zvekufamba kwembeu uye maitiro e agglutination. Brillard et al13 vakashuma kuti avhareji yehurefu hwematubule ekuchengetera sperm muvana vakuru i400–600 µm, asi mamwe maSST anogona kureba kusvika 2000 µm. Mero naOgasawara14 vakapatsanura ma seminiferous glands kuita matubules ekuchengetera mbeu akakura uye asina kukura, ese ari maviri aive akaenzana pakureba (~500 µm) uye upamhi hwemutsipa (~38 µm), asi avhareji ye lumen dhayamita yematubules yaive 56.6 uye 56.6 µm. . , zvichiteerana 11.2 μm, zvichiteerana. Muchidzidzo chazvino, takashandisa mudziyo we microfluidic une saizi yechiteshi che200 µm × 20 µm (W × H), une chikamu chakati dyo nechakawedzerwa cheSST. Pamusoro pezvo, takaongorora kufamba kwembeu uye maitiro ekubatanidza mbeu mumvura inoyerera, zvinoenderana nefungidziro yaForeman yekuti mvura inogadzirwa nemasero eSST epithelial inochengetedza mbeu mu lumen munzira inopesana ne rheological (rheological).
Chinangwa chechidzidzo ichi chaive chekukunda matambudziko ekuona kufamba kwembeu dzemurume muchubhu yefallopian uye kudzivirira matambudziko ekudzidza rheology uye maitiro embeu dzemurume munzvimbo inochinja-chinja. Mudziyo we microfluidic wakashandiswa unogadzira kumanikidzwa kwe hydrostatic kutevedzera kufamba kwembeu dzemurume munhengo dzehuku.
Kana donhwe resamu rembeu rakasanganiswa (1:40) rakaiswa mumudziyo we microchannel, mhando mbiri dzekufamba kwembeu dzaigona kuonekwa (mbeu yakaparadzaniswa nembeu yakasungwa). Pamusoro pezvo, mbeu yaiwanzo shambira ichipesana nemhepo (positive rheology; vhidhiyo 1, 2). Kunyangwe maburi embeu aive nekumhanya kwakaderera pane kwembeu isina munhu (p < 0.001), akawedzera huwandu hwembeu inoratidza rheotaxis yakanaka (p < 0.001; Tafura 2). Kunyangwe maburi embeu aive nekumhanya kwakaderera pane kwembeu isina munhu (p < 0.001), akawedzera huwandu hwembeu inoratidza rheotaxis yakanaka (p < 0.001; Tafura 2). Хотя пучки сперматозоидов имели более низкую скорость, чем у одиночных сперматозоидов (p <0,001), они увеличивали процент, сперматозоидов демонстрирующих положительный реотаксис (p <0,001; таблица 2). Kunyangwe ma spermatozoa bundles aive nespeed yakaderera pane ye single spermatozoa (p < 0.001), akawedzera huwandu hwespermatozoa hunoratidza 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美生的美美。 0.001;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 流发(p <0.001 ; 2...."...)）)) Хотя скорость пучков сперматозоидов была ниже, чем у одиночных сперматозоидов (p <0,001), они увеличивали процентозложение процент спедроман реологией (p <0,001; таблица 2). Kunyangwe kumhanya kwemasumbu embeu kwakaderera pane kwembeu imwe chete (p < 0.001), vakawedzera huwandu hwembeu ne rheology yakanaka (p < 0.001; Tafura 2).Kuvapo kwembeu imwe chete nembeu imwe chete kunofungidzirwa kuti kuri pakati pe53% uye 85%, zvichiteerana.
Zvakaonekwa kuti mbeu yehuku dzesharkasi mushure mekubuda kwembeu dzinoumba mabundles akatsetseka, ane vanhu vakawanda. Mabundles aya anowedzera kureba nekukora nekufamba kwenguva uye anogona kugara ari mu vitro kwemaawa akati wandei asati anyangarika (vhidhiyo 3). Mabundles aya ane filamentous akaumbwa se echidna spermatozoa inoumbwa pamucheto we epididymis. Hurume hweSharkashi hwakaonekwa kuti hune katsika kekuungana uye kugadzira bundle reticulate mumaminitsi asingasviki imwe mushure mekuunganidzwa. Matanda aya anoshanduka uye anokwanisa kunamira pamadziro ari pedyo kana zvinhu zvisingachinji. Kunyangwe mabundles embeu anoderedza kumhanya kwemasero embeu, zviri pachena kuti macroscopically anowedzera linearity yawo. Kureba kwemabundles kunosiyana zvichienderana nehuwandu hwembeu yakaunganidzwa mumabundles. Zvikamu zviviri zvebundle zvakaparadzaniswa: chikamu chekutanga, kusanganisira musoro wakasununguka wembeu yakasanganiswa, uye chikamu chekupedzisira, kusanganisira muswe nemagumo ese embeu. Uchishandisa kamera inomhanya zvikuru (950 fps), misoro yakasununguka yembeu dzakasanganiswa yakaonekwa muchikamu chekutanga chembeu, ichikonzera kufamba kwembeu nekuda kwekufamba kwayo, ichidhonza yasara mumbeu nekufamba kwakatenderedza (Vhidhiyo 4). Zvisinei, mumatanda marefu, zvakaonekwa kuti mimwe misoro yembeu yakasununguka yainamatira mumuviri uye chikamu chekupedzisira chembeu chinoshanda semavane ekubatsira kusunda mbeu.
Paunenge uchiyerera zvishoma nezvishoma, masumbu embeu anofamba akaenzana, zvisinei, anotanga kupindirana uye kunamatira kune zvese zvakanyarara, kuitira kuti asasukwe nekuyerera kwemvura sezvo kumhanya kwekuyerera kuchiwedzera. Masumbu acho anoumbwa kana masero mashoma embeu aswedera pedyo, anotanga kufamba akabatana uye kutenderedzana, uye obva anamatira kune chinhu chinonamira. Mifananidzo 1 ne2 inoratidza kuti mbeu inoswedera pedyo sei, ichiumba muganho apo miswe inotenderedzana.
Vaongorori vakashandisa simba remvura kuti vagadzire kuyerera kwemvura mu microchannel kuti vadzidze nezve rheology yembeu. Microchannel ine saizi ye200 µm × 20 µm (W × H) uye kureba kwe3.6 µm yakashandiswa. Shandisa microchannels pakati pezvirongo zvine sirinji dzakaiswa kumagumo. Kupenda chikafu kwakashandiswa kuti nzira dzionekwe zviri nani.
Sungai tambo dzekubatanidza nezvimwe zvinhu pamadziro. Vhidhiyo yacho yakatorwa ne phase contrast microscope. Nemufananidzo wega wega, phase contrast microscopy uye mifananidzo yemepu zvinoratidzwa. (A) Kubatana pakati penzizi mbiri kunodzivirira kuyerera nekuda kwekufamba kwehelical (museve mutsvuku). (B) Kubatana pakati pechubhu bundle nemadziro echiteshi (miseve mitsvuku), panguva imwe chete zvakabatana kune mamwe mabundle maviri (miseve yeyero). (C) Mabundle embeu ari mu microfluidic channel anotanga kubatana (miseve mitsvuku), achigadzira mesh yemabundle embeu. (D) Kuumbwa kwenetwork yemabundle embeu.
Kana donhwe rembeu rakasanganiswa nemvura rakaiswa mumudziyo we microfluidic uye mvura yakabuda, danda rembeu rakaonekwa richipesana nekwainoenda mvura. Mapango acho anowirirana zvakanaka pamadziro ema microchannels, uye misoro yakasununguka muchikamu chekutanga chemapango acho yakabatana zvakanaka paari (vhidhiyo 5). Anonamatirawo kune chero zvidimbu zvisina kumira munzira yawo, zvakaita semarara, kuti adzivise kutorwa nemvura. Nekufamba kwenguva, matuft aya anova tambo refu dzinobata dzimwe mbeu imwe chete uye matuft mapfupi (Vhidhiyo 6). Sezvo mvura inotanga kuderera, mitsetse mirefu yembeu inotanga kuumba network yemitsetse yembeu (Vhidhiyo 7; Mufananidzo 2).
Pakumhanya kwakanyanya (V > 33 µm/s), kufamba kwetambo kunowedzerwa senzira yekubata mbeu dzakawanda dzinoumba masumbu kuti dzikwanise kurwisa simba rekuyerera kwemvura. Pakumhanya kwakanyanya (V > 33 µm/s), kufamba kwetambo kunowedzerwa senzira yekubata mbeu dzakawanda dzinoumba masumbu kuti dzikwanise kurwisa simba rekuyerera kwemvura. При высокой скорости потока (V > 33 мкм/с) спиралевидные движения нитей усиливаются, поскольку они пытаются поймать множествоы озденье образующих пучки, которые лучше противостоят дрейфующей силе потока. Pakuyerera kwakanyanya (V > 33 µm/s), kufamba kwetambo dzemuchina kunowedzera sezvo dzichiedza kubata mbeu dzakawanda dzinoumba masumbu anokwanisa kudzivirira simba rekuyerera kwemvura.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精子，从而更好地抵抗流动的。在 高 流速 (v> 33 µm/s) 时, 的 螺旋 运动 增加抵抗 的 漂移力。。。。。. При высоких скоростях потока (V > 33 мкм/с) пиральное движение нитей увеличивается в попытке захватить множество отделььное движение нитей увеличивается в попытке захватить множество отделььное движение нитей увеличивается пучки, чтобы лучше сопротивляться силам дрейфа потока. Pakuyerera kwakanyanya (V > 33 µm/s), kufamba kwetambo dzesimbi kunokwira mukuyedza kubata mapete akawanda embeu kuti akwanise kudzivirira simba rekuyerera kwembeu.Vakaedzawo kubatanidza ma microchannels kumadziro emativi.
Maponde embeu akaonekwa semapoka emisoro yembeu nemiswe yakakombama uchishandisa microscopy (LM). Maponde embeu ane maponde akasiyana-siyana akaonekwawo semisoro yakakombama nemitsara yeflagellar, miswe yembeu yakawanda yakasanganiswa, misoro yembeu yakabatana nemuswe, uye misoro yembeu ine maponde akakombama semaponde akawanda akasanganiswa. transmission electron microscopy (TEM). Scanning electron microscopy (SEM) yakaratidza kuti maponde embeu aive sheathed aggregates emisoro yembeu uye maponde embeu akaratidza network yakabatana yemiswe yakaputirwa.
Maumbirwo uye maumbirwo espermatozoa, kuumbwa kwespermatozoa bundles zvakadzidzwa pachishandiswa light microscopy (hafu yechikamu), scanning electron microscopy (SEM) uye transmission electron microscopy (TEM), sperm smears dzakaiswa ruvara rweacridine orange uye dzakaongororwa pachishandiswa epifluorescence microscopy.
Kutsvukisa mbeu yemurume neacridine orange (Mufananidzo 3B) kwakaratidza kuti misoro yembeu yemurume yakanga yakabatana uye yakafukidzwa nezvinhu zvinobudisa tsvina, izvo zvakaita kuti pave nematumbu makuru (Mufananidzo 3D). Matumbu embeu yemurume aiumbwa nenhengo dzembeu yemurume dzine network yemiswe yakabatana (Mufananidzo 4A-C). Matumbu embeu yemurume anoumbwa nemiswe yembeu yemurume yakawanda yakabatana (Mufananidzo 4D). Zvakavanzika (Mufananidzo 4E,F) zvakafukidza misoro yembeu yemurume.
Kuumbwa kwenhengo yespermatozoa Kushandisa phase contrast microscopy uye sperm smears dzakapendwa neacridine orange, zvakaratidza kuti misoro yespermatozoa inonamatirana. (A) Kuumbwa kwekutanga kwesperm tuft kunotanga nesperm (denderedzwa jena) uye sperm nhatu (denderedzwa yero), ne spiral inotangira kumuswe ichipera kumusoro. (B) Photomicrograph yesperm smear yakapendwa neacridine orange inoratidza misoro yesperm inonamatira (miseve). Discharge inofukidza misoro. Kukura × 1000. (C) Kugadzirwa kwedanda guru rinotakurwa nekuyerera mu microfluidic channel (ichishandisa kamera inomhanya kwazvo pa950 fps). (D) Micrograph yesperm smear yakapendwa neacridine orange inoratidza tufts huru (miseve). Kukura: ×200.
Kuskena maelektron micrograph embeu yembeu uye smear yembeu yakasanganiswa neacridine orange. (A, B, D, E) maelektron micrographs espermatozoa ane mavara edhijitari, uye C na F maekrographs espermatozoa ane mavara eacridine orange anoratidza kubatana kwespermatozoa yakawanda ichiputira caudal web. (AC) Ma aggregates embeu anoratidzwa se network yemiswe yakabatana (miseve). (D) Kunamatira kwembeu dzakawanda (nechinhu chinonamira, mupendero wepingi, museve) zvichitenderedza muswe. (E na F) Ma aggregates emusoro wembeu (zvinongedzo) akafukidzwa nezvinhu zvinonamira (zvinongedzo). Mbeu dzakaita mabundles ane maumbirwo akafanana nevortex (F). (C) ×400 uye (F) ×200 magnifications.
Tichishandisa transmission electron microscopy, takaona kuti ma sperm bundles aive ne tails dzakanamirwa (Mufananidzo 6A, C), misoro yakanamirwa ku tails (Mufananidzo 6B), kana misoro yakanamirwa ku tails (Mufananidzo 6D). Misoro ye spermatozoa iri mu bundle yakakombama, ichiratidzwa muchikamu chechipiri che nuclear regions (Mufananidzo 6D). Mu incision bundle, spermatozoa yaive ne head yakakombama ine nucleus regions mbiri uye flagellar regions dzakawanda (Mufananidzo 5A).
Micrograph yemaelektroni ine ruvara rwedhijitari inoratidza miswe inobatanidza munhengo yembeu uye zvinhu zvinobatanidza zvinobatanidza misoro yembeu. (A) Muswe wakabatana wembeu dzakawanda. Cherechedza kuti muswe wacho unotaridzika sei mumifananidzo (museve) uye nharaunda (museve). (B) Musoro (museve) wembeu wakabatana nemuswe (museve). (C) Miswe yembeu (miseve) yakati wandei yakabatana. (D) Zvinhu zvinobatanidza (AS, bhuruu) zvinobatanidza misoro mina yembeu (yepepuru).
Kuskena electron microscopy kwakashandiswa kuona misoro yembeu muzvikamu zvembeu zvakafukidzwa nezvinobuda kana ma membrane (Mufananidzo 6B), zvichiratidza kuti zvikamu zvembeu zvakabatanidzwa nezvinhu zviri kunze kwemuviri. Zvinhu zvakasanganiswa zvakaiswa mumusoro wembeu (jellyfish head-like assembly; Mufananidzo 5B) uye zvakatambanudzwa kure, zvichipa chitarisiko cheyero chakajeka pasi pe fluorescence microscopy kana zvaiswa acridine orange (Mufananidzo 6C). Chinhu ichi chinoonekwa zvakajeka pasi pe scanning microscope uye chinoonekwa sechinobatanidza. Zvikamu zvakatetepa zvishoma (Mufananidzo 5C) uye sperm smears dzakaiswa acridine orange dzakaratidza zvikamu zvembeu zvine misoro yakazara uye miswe yakakombama (Mufananidzo 5D).
Mifananidzo yakasiyana-siyana inoratidza kuunganidzwa kwemisoro yembeu nemiswe yakapetwa uchishandisa nzira dzakasiyana-siyana. (A) Micrograph yemagetsi inotumirwa nemavara edhijitari ine mutsetse wembeu inoratidza musoro wembeu wakakombama une nucleus ine zvikamu zviviri (bhuruu) uye zvikamu zvakati wandei zveflagellar (girini). (B) Micrograph yemagetsi inoskena nemavara edhijitari inoratidza boka remisoro yembeu yakaita sejellyfish (miseve) inoita kunge yakafukidzwa. (C) Chikamu chakatetepa chinoratidza misoro yembeu yakaunganidzwa (miseve) nemiswe yakakombama (miseve). (D) Micrograph yembeu yakasanganiswa neacridine orange inoratidza misoro yembeu (miseve) nemiswe yakakombama (miseve). Cherechedza kuti chinhu chinonamira (S) chinofukidza musoro wembeu. (D) × 1000 magnification.
Kushandisa transmission electron microscopy (Mufananidzo 7A), zvakaonekwawo kuti misoro yembeu yakanga yakakombama uye manyuchi aiva nechimiro che spiral, sezvakasimbiswa ne sperm smears dzakapendwa ne acridine orange uye dzakaongororwa uchishandisa fluorescence microscopy (Mufananidzo 7B).
(A) Micrograph yemagetsi inotumirwa neruvara rwedhijitari uye (B) Acridine orange stained sperm smear inoratidza misoro yakakombama uye kubatana kwemisoro yesperm nemiswe (miseve). (B) × 1000 magnification.
Chimwe chinhu chinonakidza chakawanikwa ndechekuti mbeu yaSharkazi inoungana kuti igadzire zvidimbu zvinofamba zvine filamentous bundles. Hunhu hwezvikamu izvi hunotibatsira kunzwisisa basa razvo mukutora nekuchengetera mbeu yemurume muSST.
Mushure mekusangana kwembeu, mbeu yembeu inopinda munhengo yemukadzi uye inoitwa sarudzo yakasimba, zvichikonzera huwandu hushoma hwembeu hunopinda muSST15,16. Kusvika pari zvino, nzira dzinopinda nekubuda nayo mbeu yembeu muSST hadzina kujeka. Muhuku, mbeu yembeu inochengetwa muSST kwenguva yakareba yemavhiki maviri kusvika gumi, zvichienderana nerudzi rwayo6. Gakava riripo pamusoro pemamiriro embeu panguva yekuchengetwa muSST. Dziri kufamba here kana kuti dzakazorora? Nemamwe mashoko, mbeu yembeu inochengetedza sei nzvimbo yayo muSST kwenguva yakareba kudaro?
Forman4 akataura kuti kugara kweSST nekubuda kwembeu zvinogona kutsanangurwa maererano nekufamba kwembeu. Vanyori vanofungidzira kuti mbeu inochengetedza nzvimbo yayo nekushambira ichipesana nekuyerera kwemvura kunogadzirwa neSST epithelium uye kuti mbeu inoburitswa kubva kuSST kana kumhanya kwayo kwaderera pasi penzvimbo yainotanga kudzokera shure nekuda kwekushaya simba. Zaniboni5 yakasimbisa kuvapo kweaquaporins 2, 3, uye 9 muchikamu cheapical chemasero eSST epithelial, izvo zvinogona kutsigira nenzira isina kunanga modhi yekuchengetera mbeu yaForeman. Muchidzidzo chazvino, takaona kuti inenge hafu yembeu yaSharkashi inoratidza rheology yakanaka mumvura inoyerera, uye kuti mabundle embeu akabatanidzwa anowedzera huwandu hwembeu inoratidza rheology yakanaka, kunyangwe agglutination ichidzikisa kumhanya kwawo. Mafambiro anoita mbeu muchubhu yeshiri kuenda kunzvimbo yekuberekesa haanzwisisike zvizere. Mumhuka dzinoyamwisa, fluid yefollicular chemoattracts spermatozoa. Zvisinei, chemoattractants inofungidzirwa kuti inotungamira mbeu kusvika kure7. Saka, mamwe maitiro ndiwo anokonzera kutakurwa kwembeu. Kugona kwembeu kutenderera nekuyerera ichipesana nemvura yemuchubhu yefallopian inoburitswa mushure mekusangana kwakataurwa kuti ndicho chinhu chikuru pakutarisa mbeu mumbeu. Parker 17 akataura kuti mbeu yembeu inoyambuka maoviducts nekushambira ichipesana nemhepo yeciliary mushiri nezvinokambaira. Kunyangwe zvisati zvaratidzwa mukuyedza mushiri, Adolphi18 ndiye akatanga kuona kuti mbeu yembeu yeshiri inopa mhedzisiro yakanaka kana jira rakatetepa remvura pakati pechivharo nechitsvedza rakagadzirwa nepepa rekusefa. Rheology. Hino naYanagimachi [19] vakaisa ovary-tubal-uterine complex yembeu mumhete yekuyerera kwemvura ndokuisa 1 µl yeinki muisthmus kuti vaone kuyerera kwemvura muchubhu yefallopian. Vakaona kufamba kwakasimba kwekudzikira nekureruka muchubhu yefallopian, umo mabhora ese einki aifamba zvishoma nezvishoma achienda kuampulla yechubhu yefallopian. Vanyori vanosimbisa kukosha kwekuyerera kwemvura yemuchubhu kubva pasi kuenda kuchubhu yepamusoro yefallopian kuti mbeu ikwire nekubereka. Brillard20 yakashuma kuti muhuku neturkeys, mbeu dzinofamba nekufamba dzichibva pamusuwo wemukadzi, kwadzinochengeterwa, kuenda kunosangana kwemukadzi nemukadzi, kwadzinochengeterwa. Zvisinei, kufamba uku hakudiwi pakati pekubatana kwemukadzi ne infundibulum nekuti mbeu dzinofambiswa nekufamba kwemukadzi. Tichiziva mazano aya ekare uye mhedzisiro yakawanikwa muchidzidzo chazvino, zvinogona kufungidzirwa kuti kugona kwembeu kufamba kumusoro (rheology) ndeimwe yehunhu hwakavakirwa pakusarudza. Izvi zvinosarudza kupfuura kwembeu nemukadzi uye kupinda kwayo muCCT kuti ichengetwe. Sezvakataurwa naForman4, izvi zvinogona zvakare kubatsira maitiro embeu kupinda muSST nenzvimbo yayo kwenguva yakati uye yozobuda kana kumhanya kwayo kwatanga kudzikira.
Kune rumwe rutivi, Matsuzaki naSasanami 21 vakataura kuti mbeu yembeu yeshiri inochinja kufamba kubva pakurara kuenda pakugona kufamba munhengo dzekubereka dzevarume nedzevakadzi. Kudziviswa kwekufamba kwembeu yembeu yembeu muSST kwakakurudzirwa kutsanangura nguva yakareba yekuchengetwa kwembeu uye kudzorerwa mushure mekubuda muSST. Mumamiriro ezvinhu asina oxygen, Matsuzaki nevamwe vake vakataura nezvekuburitswa kwakanyanya kwe lactate muSST, izvo zvinogona kutungamira mukudziviswa kwekufamba kwembeu yembeu yembeu yembeu. Muchiitiko ichi, kukosha kwe rheology yembeu kunoratidzwa mukusarudza nekunwa kwembeu yembeu, kwete mukuchengetedzwa kwayo.
Maitiro ekubatana kwembeu anoonekwa setsananguro inonzwisisika yenguva refu yekuchengetwa kwembeu muSST, sezvo iyi iri nzira yakajairika yekuchengetedza mbeu muhuku2,22,23. Bakst et al. 2 vakaona kuti mbeu zhinji dzainamatirana, dzichigadzira fascicular aggregates, uye mbeu imwe chete yaisawanzo kuwanikwa muCCM yezvihuta. Kune rumwe rutivi, Wen et al. 24 vakaona mbeu dzakawanda dzakapararira uye mbeu shoma dzembeu muSST lumen muhuku. Zvichibva pane izvi, zvinogona kufungidzirwa kuti mukana wekubatana kwembeu unosiyana pakati peshiri nembeu muhuku imwe chete. Pamusoro pezvo, Van Krey et al. 9 vakataura kuti kupatsanurwa kwembeu dzakasanganiswa ndiko kunokonzera kupinda kwembeu zvishoma nezvishoma mulumen yefallopian tube. Zvichienderana nepfungwa iyi, mbeu dzine simba rakaderera rekubatana dzinofanira kuburitswa muSST kutanga. Munyaya iyi, kugona kwembeu kuungana kunogona kunge kuri chinhu chinopesvedzera mhedzisiro yekukwikwidzana kwembeu mushiri dzine tsvina. Pamusoro pezvo, kana mbeu yakaungana ikaramba ichingopatsanuka, kubereka kunoenderera mberi kwenguva refu.
Kunyangwe kuungana kwembeu dzemudumbu nekuungana muzvikamu zvakaonekwa muzvidzidzo zvakati wandei2,22,24, hazvina kutsanangurwa zvakadzama nekuda kwekuoma kwekutarisa kwadzo mukati meSST. Kuedza kwakawanda kwakaitwa kudzidza kuungana kwembeu dzemudumbu mu vitro. Kuungana kwakakura asi kwenguva pfupi kwakaonekwa apo waya yakatetepa yakabviswa kubva pakudonha kwembeu yakarembera. Izvi zvinotungamira kukuti bubble refu rinobuda kubva pakudonha, richitevedzera gland rembeu. Nekuda kwemiganhu ye3D uye nguva pfupi yekuomesa madonhwe, bhokisi rese rakakurumidza kudonha9. Muchidzidzo chazvino, tichishandisa huku dzeSharkashi nemachipisi emicrofluidic, takakwanisa kutsanangura kuti matufts aya anoumbwa sei uye kuti anofamba sei. Zvikamu zvembeu zvemudumbu zvakaumbwa pakarepo mushure mekuunganidzwa kwembeu uye zvakawanikwa zvichifamba mudenderedzwa, zvichiratidza rheology yakanaka kana iripo mukuyerera. Uyezve, pazvakaonekwa nemacroscopically, zvikamu zvembeu zvemudumbu zvakaonekwa zvichiwedzera linearity yekufamba zvichienzaniswa nembeu dzakaparadzana. Izvi zvinoratidza kuti kuungana kwembeu yemurume kunogona kuitika SST isati yapinda uye kuti kugadzirwa kwembeu yemurume hakungogumiri panzvimbo diki nekuda kwekunetseka sezvakambotaurwa (Tingari neLake12). Panguva yekuumbwa kwetuft, mbeu yemurume inoshambira mu synchrony kusvika yaita junction, ipapo miswe yayo inotenderera uye musoro wembeu yemurume unoramba wakasununguka, asi muswe nechikamu chedistal chembeu yemurume zvinonamira pamwe chete nechinhu chinonamira. Nokudaro, musoro wembeu yemurume ndiyo inotungamira kufamba, ichidhonza mamwe masero embeu yemurume. Kuskena electron microscopy yema sperm bundles kwakaratidza misoro yembeu yakabatana yakafukidzwa nezvinhu zvakawanda zvinonamira, zvichiratidza kuti misoro yembeu yemurume yakabatanidzwa muma bundles akareruka, izvo zvingave zvakaitika mushure mekusvika panzvimbo yekuchengetera (SST).
Kana smear yembeu yatsvukiswa neacridine orange, zvinhu zvinonamira kunze kwemasero embeu zvinogona kuonekwa uchishandisa maikorosikopu ine fluorescent. Chinhu ichi chinobvumira mabundle embeu kunamira nekunamatira kune chero nzvimbo dzakatenderedza kana zvidimbu kuitira kuti zvisafambe nekuyerera kwakapoteredza. Saka, zvatakaona zvinoratidza basa rekunamatira kwembeu muchimiro chemabundle anofambiswa. Kugona kwavo kushambira vachipesana nemhepo uye kunamatira kunzvimbo dziri pedyo kunoita kutimbeu igare kwenguva refu muSST.
Rothschild25 akashandisa kamera ye hemocytometry kuti adzidze kupararira kwembeu yemombe mudonhwe re suspension, achitora ma photomicrographs kuburikidza nekamera ine vertical ne horizontal optical axis ye microscope. Zvakabuda zvakaratidza kuti spermatozoa yakakwezvwa pamusoro pekamuri. Vanyori vanoti panogona kunge paine hydrodynamic interactions pakati pembeu nepamusoro. Tichifunga izvi, pamwe nekukwanisa kwembeu yeSharkashi chick kugadzira tufts dzinonamira, zvinogona kuwedzera mukana wekuti mbeu irambe yakanamatira pamadziro eSST uye ichengetwe kwenguva yakareba.
Bccetti naAfzeliu26 vakataura kuti glycocalyx yembeu inodiwa kuti gamete izivikanwe uye iunganidzwe. Forman10 vakaona kuti hydrolysis ye α-glycosidic bonds mu glycoprotein-glycolipid coatings nekurapa mbeu yembeu ne neuraminidase zvakakonzera kudzikira kwekubereka pasina kukanganisa kufamba kwembeu. Vanyori vanoti mhedzisiro ye neuraminidase pa glycocalyx inokanganisa kuunganidzwa kwembeu pa utero-vaginal junction, nokudaro ichideredza kubereka. Zvavakaona hazvigone kufuratira mukana wekuti kurapwa neuraminidase kunogona kuderedza kuzivikanwa kwembeu ne oocyte. Forman naEngel10 vakaona kuti kubereka kwakadzikira apo huku dzaiiswa mbeu mukati memukadzi nembeu dzakarapwa ne neuraminidase. Zvisinei, IVF nembeu dzakarapwa ne neuraminidase haina kukanganisa kubereka zvichienzaniswa nehuku dzinodzora. Vanyori vakagumisa kuti shanduko mu glycoprotein-glycolipid coating yakakomberedza membrane yembeu yakaderedza kugona kwembeu kusanganisa nekukanganisa kuunganidzwa kwembeu panosangana utero-vaginal, izvo zvakazoita kuti kurasikirwa kwembeu kuwedzere nekuda kwekumhanya kwekubatanidza utero-vaginal, asi hazvikanganisi kuzivikanwa kwembeu nemazai.
Mumaturkeys, Bakst naBauchan 11 vakawana zvidimbu zvidiki zvema vesicles ne membrane mu lumen yeSST uye vakaona kuti mamwe ema granules aya akanga akabatana ne sperm membrane. Vanyori vanoti hukama uhwu hunogona kubatsira pakuchengetwa kwenguva refu kwespermatozoa muSST. Zvisinei, vaongorori havana kutaura kwakabva zvidimbu izvi, kungave zvakaburitswa neCCT epithelial cells, zvinogadzirwa uye zvinoburitswa nesystem yekubereka yevarume, kana kuti zvinoburitswa nesperm pachayo. Zvakare, zvidimbu izvi zvine basa re agglutination. Grützner et al27 vakataura kuti epididymal epithelial cells dzinogadzira uye dzinoburitsa protein chaiyo inodiwa pakuumbwa kwema single-pore seminal tracts. Vanyori vanotaurawo kuti kupararira kwema bundles aya kunoenderana nekudyidzana kwema epididymal proteins. Nixon et al28 vakaona kuti adnexa inoburitsa protein, acidic cysteine-rich osteonectin; SPARC inobatanidzwa mukuumbwa kwema sperm tufts mu echidnas ne platypuses dzine muromo mupfupi. Kupararira kwemavara aya kunobatanidzwa nekurasikirwa kwepuroteni iyi.
Muongororo yazvino, ongororo ye ultrastructural uchishandisa electron microscopy yakaratidza kuti spermatozoa yainamatira kune zvinhu zvakawanda zvakakora. Zvinhu izvi zvinofungidzirwa kuti ndizvo zvinokonzera agglutination inoungana pakati nepakati pemisoro inonamatira, asi paine huwandu hwakaderera munharaunda yemuswe. Tinofunga kuti chinhu ichi chinounganidza chinoburitswa kubva munhengo yekubereka yevarume (epididymis kana vas deferens) pamwe chete nembeu, sezvo tichiwanzoona urume huchiparadzana ne lymph ne seminal plasma panguva yekubuda kwembeu. Zvakataurwa kuti sezvo avian spermatozoa inopfuura nepakati pe epididymis ne vas deferens, inosangana nekuchinja kwakabatana nekukura kunotsigira kugona kwayo kusunga mapuroteni nekuwana plasma lemma-associated glycoproteins. Kuramba kuripo kwemapuroteni aya pa resident sperm membranes muSST kunoratidza kuti mapuroteni aya anogona kukanganisa kuwanikwa kwe sperm membrane stability 30 uye kusarudza kubereka kwavo 31. Ahammad nevamwe vake32 vakataura kuti mbeu dzemurume dzinowanikwa kubva muzvikamu zvakasiyana-siyana zvenhengo dzekubereka dzevarume (kubva kumatestes kusvika kuma distal vas deferens) dzakaratidza kuwedzera kuri kuita hupenyu hwemhuka pasi pemamiriro ekuchengetwa kwemvura, pasinei nekupisa kwenzvimbo yekuchengetwa, uye hupenyu hwemhuka hunowedzerawo mumachubhu efallopian mushure mekuisirwa mbeu dzechivanhu.
Matuft embeu dzehuku dzeSharkashi ane hunhu hwakasiyana uye mabasa akasiyana neamwe marudzi akadai seechidnas, platypuses, wood mice, deer rats, uye Guinea pigs. Muhuku dzeSharkasi, kuumbwa kwembeu dzehuku dzakaderedza kumhanya kwadzo kwekushambira zvichienzaniswa nembeu dzehuku imwe chete. Zvisinei, matuft aya akawedzera huwandu hwembeu dzehuku dzine rheologically positive uye akawedzera kugona kwembeu dzehuku kuti dzigadzikane munzvimbo ine simba. Saka, zvatakawana zvinosimbisa fungidziro yapfuura yekuti kuungana kwembeu muSST kunobatanidzwa nekuchengetwa kwembeu kwenguva refu. Tinofungidzirawo kuti tsika yembeu yekuumba tufts inogona kudzora mwero wekurasikirwa kwembeu muSST, izvo zvinogona kuchinja mhedzisiro yekukwikwidzana kwembeu. Sekufunga uku, mbeu dzehuku dzine agglutination shoma dzinoburitsa SST kutanga, nepo mbeu dzehuku dzine agglutination yakawanda dzinoburitsa vana vazhinji. Kuumbwa kwembeu dzehuku dzehuku dzine pore imwe chete kunobatsira uye kunokanganisa chiyero chemubereki nemwana, asi kunoshandisa nzira yakasiyana. Muma echidnas nema platypuses, mbeu dzemukomana dzakarongwa dzakafanana kuti dziwedzere kumhanya kwemuti. Maponde e echidnas anofamba nekukurumidza katatu kupfuura mbeu imwe chete. Zvinotendwa kuti kuumbwa kwembeu dzakadaro mu echidnas inzira yekushanduka-shanduka kuti irambe ichitonga, sezvo vanhukadzi vachigara vachisangana nevanhurume vakawanda uye vanowanzosangana nevanhurume vakawanda. Saka, mbeu dzemukomana dzakasiyana dzinokwikwidzana zvakasimba kuti zai ribereke.
Kuongororwa kwembeu dzehuku dzesharkasi kuri nyore kuona uchishandisa phase contrast microscopy, iyo inoonekwa seyakanakira nekuti inobvumira kudzidza zviri nyore maitiro embeu dzehuku mu vitro. Maitiro ayo kuumbwa kwembeu dzehuku kunokurudzira kubereka muhuku dzesharkasi akasiyanawo neanowanikwa mune dzimwe mhuka dzinoyamwisa dzemudumbu dzinomiririra maitiro embeu dzemudumbu akadai sembeva dzehuni, uko dzimwe mbeva dzinosvika mazai, zvichibatsira vamwe vanhu vane hukama kusvika nekukuvadza mazai avo. kuratidza kuti uri wega. maitiro ekuzvipira. Kuzviberekesa 34. Mumwe muenzaniso wemaitiro ekushandira pamwe mumbeu dzehuku wakawanikwa mumbeva dzemhara, uko mbeva dzakakwanisa kuziva nekubatana nembeu dzemudumbu dzine hukama hwakanyanya uye kuumba mapoka ekushandira pamwe kuti dziwedzere kumhanya kwadzo zvichienzaniswa nembeu dzisina hukama 35.
Zvakawanikwa muchidzidzo ichi hazvipikisi dzidziso yaFoman yekuchengetwa kwenguva refu kwembeu dzemudumbu muSWS. Vaongorori vanoti masero embeu anoramba achifamba mukufamba kwemasero epithelial ari mukati meSST kwenguva yakareba, uye mushure menguva yakati, matura esimba embeu dzemudumbu anodzikira, zvichikonzera kudzikira kwekumhanya, izvo zvinobvumira kuburitswa kwesimba rembeu dzemudumbu nekuyerera kwemvura kubva mu lumen yeSST. Muchidzidzo chazvino, takaona kuti hafu yembeu imwe chete yakaratidza kugona kushambira ichipesana nemvura inoyerera, uye kunamatira kwayo mubundle kwakawedzera kugona kwayo kuratidza rheology yakanaka. Uyezve, data redu rinoenderana neraMatsuzaki et al. 1 avo vakataura kuti kuwedzera kwe lactate secretion muSST kunogona kudzivirira kufamba kwembeu dzemudumbu. Zvisinei, zvatakawana zvinotsanangura kuumbwa kwembeu dzemudumbu dzinofamba uye maitiro adzo erheological pamberi penzvimbo inoshanduka mukati me microchannel mukuedza kujekesa maitiro avo muSST. Ongororo yeramangwana inogona kutarisa pakuona kuti makemikari api uye kwakabva chinhu chinosanganisa masero, izvo zvichabatsira vaongorori kugadzira nzira itsva dzekuchengetedza urume hwemvura uye kuwedzera nguva yekubereka.
Shiri gumi neshanu dzine sharkasi (homozygous dominant; Na Na) dzine mavhiki makumi matatu ekuberekwa dzakasarudzwa sevanopa mbeu dzembeu muongororo iyi. Shiri idzi dzakarerwa paResearch Poultry Farm yeFaculty of Agriculture, Ashit University, Ashit Governorate, Egypt. Shiri dzakachengetwa mumakeji akasiyana (30 x 40 x 40 cm), dzakaiswa chiedza (maawa gumi nematanhatu echiedza uye maawa masere erima) uye dzakapihwa chikafu chine 160 g yeprotein isina kucheneswa, 2800 kcal yesimba rinogadziriswa, 35 g yecalcium imwe neimwe. 5 magiramu ephosphorus inowanikwa pakirogiramu yekudya.
Sekureva kwedata 36, ​​​​37, mbeu yakatorwa kubva kuvarume kuburikidza nekukwizwa mudumbu. Masampuli embeu makumi mana nemashanu akatorwa kubva kuvarume gumi nevashanu kwemazuva matatu. Mbeu (n = 15/zuva) yakasanganiswa nekukurumidza ne1:1 (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 333 mOsm/kg38. Sampuro dzembeu dzakasanganiswa dzakatanga dzaongororwa nemaikorosikopu kuti dzionekwe kuti mbeu dzacho dzakanaka (hunyoro) uye dzozochengetwa mumvura inodziya pa37°C kusvika dzashandiswa mukati mehafu yeawa mushure mekutorwa.
Kinematics uye rheology yespermatozoa zvinotsanangurwa uchishandisa system ye microfluidic devices. Sampuli dzembeu dzakawedzerwa kuderedzwa kusvika 1:40 muBeltsville Avian Semen Diluent, dzakaiswa mu microfluidic device (ona pazasi), uye kinetic parameters dzakaongororwa uchishandisa Computerized Semen Analysis (CASA) system yakagadzirwa kare ye microfluidics characterization. pakufamba kwespermatozoa mu liquid media (Dhipatimendi reMechanical Engineering, Faculty of Engineering, Assiut University, Egypt). Plugin inogona kudhawunirodhwa pa: http://www.assiutmicrofluidics.com/research/casa39. Curve velocity (VCL, μm/s), linear velocity (VSL, μm/s) uye average trajectory velocity (VAP, μm/s) zvakayerwa. Mavhidhiyo espermatozoa akatorwa uchishandisa inverted Optika XDS-3 phase contrast microscope (ine 40x objective) yakabatana neTucson ISH1000 camera pa30 fps kwemasekondi matatu. Shandisa software yeCASA kuti udzidze nzvimbo nhatu uye nzira dzembeu mazana mashanu pamuenzaniso wega wega. Vhidhiyo yakarekodhwa yakagadziriswa uchishandisa CASA yakagadzirwa kumba. Tsanangudzo yekufamba muCASA plug-in yakavakirwa pakumhanya kwembeu kana tichienzanisa nekufamba kwayo, uye haisanganisire zvimwe zvinhu zvakaita sekufamba kwembeu padivi nepadivi, sezvo izvi zvakaonekwa kuti zvakavimbika mukuyerera kwembeu. Kufamba kwembeu kunotsanangurwa sekufamba kwembeu padivi pekufamba kwembeu. Mbeu ine hunhu hwemheu yakakamurwa nenhamba yembeu inofamba; mheu yaive yakamira uye mheu inofamba zvishoma nezvishoma haina kuverengerwa.
Makemikari ese akashandiswa akawanikwa kubva kuElgomhoria Pharmaceuticals (Cairo, Egypt) kunze kwekunge zvataurwa neimwe nzira. Mudziyo uyu wakagadzirwa sezvakatsanangurwa naEl-sherry et al. 40 pamwe nekumwe kuchinja. Zvinhu zvakashandiswa kugadzira ma microchannels zvaisanganisira ma glass plates (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 dzinogadzirwa uchishandisa soft lithography. Kutanga, mask yekudzivirira kumeso yakajeka ine dhizaini ye microchannel yaidiwa yakadhindwa paprinta ine resolution yepamusoro (Prismatic, Cairo, Egypt nePacific Arts and Design, Markham, ON). Mamasters akagadzirwa achishandisa magirazi plates se substrates. Ma plates akacheneswa ne acetone, isopropanol nemvura yakabviswa ion ndokuzoputirwa ne 20 µm layer yeSU8-25 ne spin coating (3000 rpm, 1 min). Zvikamu zveSU-8 zvakazoomeswa zvinyoro nyoro (65°C, 2 min uye 95°C, 10 min) ndokuiswa pamwaranzi yeUV kwemasekonzi makumi mashanu. Bika mushure mekupiswa pa65°C uye 95°C kwemaminitsi 1 ne4 kuti zvisanganiswe zvikamu zveSU-8, zvichiteverwa nekugadzira diacetone alcohol kwemaminitsi 6.5. Bika waffles zvakasimba (200°C kwemaminitsi 15) kuti SU-8 inyatsosimba.
PDMS yakagadzirwa nekusanganisa monomer ne hardener muchiyero chehuremu hwe10:1, yozobviswa utsi mu vacuum desiccator yodururwa pa SU-8 main frame. PDMS yakachekwa muovheni (120°C, 30 min), ipapo nzira dzakachekwa, dzakaparadzaniswa kubva ku master, uye dzakaboorwa kuti machubhu akwanise kunamirwa panzvimbo inopinzwa neinobuda ye microchannel. Pakupedzisira, nzira dze PDMS microchannel dzakanamirwa zvachose kuma microscope slides uchishandisa portable corona processor (Electro-Technic Products, Chicago, IL) sezvakatsanangurwa kune dzimwe nzvimbo. Microchannel yakashandiswa muchidzidzo ichi inoyera 200 µm × 20 µm (W × H) uye yakareba 3.6 cm.
Kuyerera kwemvura kunokonzerwa nekumanikidzwa kwehydrostatic mukati me microchannel kunowanikwa nekuchengetedza huwandu hwemvura mu inlet reservoir pamusoro pemusiyano wekukwirira Δh39 mu outlet reservoir (Mufananidzo 1).
apo f iri coefficient of friction, inotsanangurwa se f = C/Re yekufamba kwe laminar muchiteshi che rectangular, apo C iri constant zvichienderana ne aspect ratio yechiteshi, L ndiyo kureba kwechiteshi che microchannel, Vav ndiyo avhareji yevelocity mukati mechiteshi che microchannel, Dh ndiyo hydraulic diameter yechiteshi, g - kukurumidza kwegiravhiti. Uchishandisa equation iyi, avhareji yevelocity yechiteshi inogona kuverengerwa uchishandisa equation inotevera: