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Uburumbuke bw'inyoni buterwa n'ubushobozi bwazo bwo kubika intanga zihagije mu gihe kirekire mu tubules zo kubika intanga (SST). Uburyo nyabwo intanga zinjira, zigaturamo, kandi zigasohoka muri SST buracyari ikibazo. Intanga z'inkoko zo mu bwoko bwa sharkasi zagaragaje ko zikunda guterana cyane, zigakora uduce twinshi tw'udusimba tugenda duhindukamo uturemangingo. Bitewe n'ingorane zo kureba uko intanga zigenda n'imyitwarire yazo mu muyoboro w'inda utagaragara, twakoresheje igikoresho cya microfluidic gifite igice gito gisa n'icya spermatozoa kugira ngo twige uburyo intanga zigenda n'uko zigenda. Ubu bushakashatsi buvuga ku buryo intanga zigenda, uburyo zigenda, n'uruhare rwazo mu kongera igihe intanga ziguma muri SST. Twakoze ubushakashatsi ku muvuduko w'intanga n'imyitwarire ya rheological iyo amazi atembera mu muyoboro w'inda aturutse ku muvuduko wa hydrostatic (igipimo cy'umuvuduko = 33 µm/s). Intanga zikunda koga zinyuranyije n'umuvuduko w'amazi (rheology nziza) kandi umuvuduko w'intanga zigenda ugabanuka cyane ugereranije n'intanga zigenda imwe. Byagaragaye ko uduce tw’intanga ngabo twimuka mu buryo buzinze kandi twiyongera uburebure n’ubugari uko intanga ngabo imwe imwe zigenda ziyongera. Uduce tw’intanga ngabo twagaragaye twegera kandi tugafata ku mpande z’imiyoboro ya microfluidic kugira ngo twirinde gutwarwa n’umuvuduko w’amazi > 33 µm/s. Uduce tw’intanga ngabo twagaragaye twegera kandi tugafata ku mpande z’imiyoboro ya microfluidic kugira ngo twirinde gutwarwa n’umuvuduko w’amazi > 33 µm/s. Было замечено, что пучки спер музозоидов приближаются и прилипают к боковым мкакам матфлюидных панов, ч чы избежать сметания со скоросту потока жидкости> 33 мкм / с. Byagaragaye ko uduce tw’intanga ngabo twegera kandi tugafata ku nkuta z’impande z’imiyoboro ya microfluidic kugira ngo tudatwarwa n’amazi atemba >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 спер музозоидов приближаются и прилипают к боковым мкакам мжидкостного канала, ч чы избежать сметания потоком жидкости со секоростю> 33 мкм / с. Byagaragaye ko intanga ngabo zegera kandi zigafata ku nkuta zo ku ruhande rw'umuyoboro wa microfluidic kugira ngo hirindwe ko amazi atemba kuri >33 µm/s.Gusuzuma no gupima amashanyarazi byagaragaje ko uduce tw’intanga ngabo twari dufite ibikoresho byinshi byuzuye. Amakuru yabonetse agaragaza uburyo spermatozoa y’inkoko ya Sharkazi igenda yiyongera, ndetse n’ubushobozi bw’intanga ngabo bwo kwihuza no gukora uduce tw’intanga ngabo, ibyo bikaba bigira uruhare mu gusobanukirwa neza uburyo spermatozoa ibikwa mu gihe kirekire muri SMT.
Kugira ngo abantu n'inyamaswa nyinshi bagere ku intanga ngabo, intanga ngabo n'amagi bigomba kugera aho intanga ngabo zibera mu gihe gikwiye. Kubwibyo, guhuza bigomba kubaho mbere cyangwa mu gihe cyo gutanga intanga ngabo. Ku rundi ruhande, zimwe mu nyamaswa, nk'imbwa, ndetse n'izindi nyamaswa zitari intanga ngabo, nk'udukoko, amafi, ibikururuka, n'inyoni, bibika intanga ngabo mu myanya yazo y'imyororokere igihe kirekire kugeza igihe amagi yazo azaba yiteguye gusama (gusama intanga ngabo 1). Inyoni zishobora gukomeza kubaho kw'intanga ngabo zishobora gusama amagi mu gihe cy'ibyumweru 2-102.
Iki ni ikintu kidasanzwe gitandukanya inyoni n'izindi nyamaswa, kuko gitanga amahirwe menshi yo gusama nyuma yo gutera intanga rimwe mu gihe cy'ibyumweru byinshi nta guhuza no gutera intanga icyarimwe. Urugingo rw'ingenzi rubika intanga, rwitwa tubule yo kubika intanga (SST), ruherereye mu mikurire y'imbere mu gice cy'uruhinja rw'intanga. Kugeza ubu, uburyo intanga zinjira, zigatura, kandi zigasohoka mu kigo cy'intanga ntiburasobanuka neza. Hashingiwe ku bushakashatsi bwakozwe mbere, hari ibitekerezo byinshi byatanzwe, ariko nta na kimwe muri byo cyemejwe.
Forman4 yatekereje ko intanga ngabo ziguma mu mwanya wazo mu bwobo bwa SST binyuze mu kugenda buhoro buhoro mu buryo bunyura mu cyerekezo cy’amazi atembera mu miyoboro ya poroteyine iherereye ku turemangingo twa epithelial twa SST (rheology). ATP iragabanuka bitewe n’imikorere ihoraho y’intanga ngabo ikenewe kugira ngo intanga ngabo zigume mu gitereko cya SST kandi kugenda kwazo kugabanuka kugeza igihe intanga ngabo zisohokeye mu bubiko bw’intanga ngabo binyuze mu gutembera kw’amazi no gutangira urugendo rushya mu muyoboro w’intanga ngore uzamuka kugira ngo inyoboze intanga ngabo. Igi (Forman4). Ubu buryo bwo kubika intanga ngabo bushyigikiwe no kumenya aquaporins 2, 3 na 9 ziboneka mu turemangingo twa epithelial twa SST. Kugeza ubu, ubushakashatsi ku intanga ngabo z’inkoko n’uruhare rwazo mu kubika SST, guhitamo intanga ngabo, no guhangana kw’intanga ngore burabura. Mu nkoko, intanga ngabo zinjira mu gitsina nyuma yo gushyingiranwa bisanzwe, ariko intanga ngabo zirenga 80% zisohoka mu gitsina nyuma gato yo gushyingiranwa. Ibi bigaragaza ko igitsina ari cyo kibanza cy’ibanze cyo gutoranya intanga ngabo mu nyoni. Byongeye kandi, byatangajwe ko munsi ya 1% by'intanga ngore zatewe mu gitsina zishyirwa mu byiciro bya SST2. Mu gutera intanga mu gitsina mu buryo bw'ubukorano, umubare w'intanga ngore zigera kuri SST ukunda kwiyongera amasaha 24 nyuma yo gutera intanga. Kugeza ubu, uburyo bwo gutoranya intanga muri iki gikorwa ntabwo busobanutse neza, kandi kugenda kw'intanga ngore bishobora kugira uruhare runini mu kwinjiza intanga ngore muri SST. Bitewe n'inkuta nini kandi zitagaragara neza z'imiyoboro y'inkoko, biragoye gukurikirana mu buryo butaziguye kugenda kw'intanga mu miyoboro y'inkoko. Kubwibyo, ntabwo dufite ubumenyi bw'ibanze ku buryo intanga ngore zihinduka SST nyuma yo gutera intanga.
Inyigisho y’indwara z’imitsi iherutse kwemerwa nk'ikintu cy'ingenzi kigenzura ubwikorezi bw'intanga mu gitsina cy'inyamabere. Hashingiwe ku bushobozi bw'intanga zigenda zimuka mu buryo bunyuranyije n'ubwo, Zaferani n'abandi bakoresheje sisitemu ya corra microfluidic kugira ngo bakuremo intanga zigenda zimuka mu masohoro yafashwe. Ubu buryo bwo gutoranya intanga ni ingenzi mu kuvura ubugumba no mu bushakashatsi ku buvuzi, kandi bukundwa kuruta uburyo gakondo butwara igihe kinini kandi bushobora kwangiza imiterere y'intanga n'imiterere yayo. Ariko, kugeza ubu, nta bushakashatsi bwakozwe ku ngaruka z'imisemburo y'inkoko ku mikorere y'intanga.
Nubwo uburyo bwo kubungabunga intanga buguma mu itera ry’imisemburo, abashakashatsi benshi babonye ko intanga zituye zihurira ku mutwe mu itera ry’imisemburo mu nkoko 9, 10, inkware 2, n’inkoko 11 kugira ngo zikore uduce tw’intanga zifatanye. Abanditsi bavuga ko hari isano iri hagati y’uku guhuzwa no kubika intanga mu gihe kirekire mu itera ry’imisemburo.
Tingari na Lake12 batangaje isano ikomeye hagati y’intanga ngabo mu gasabo k’inkoko kakira intanga ngabo, maze bibaza niba intanga ngabo z’inyoni zihuza imiterere yazo nk’uko intanga ngabo z’inyamabere zihuza. Bemeza ko isano iri hagati y’intanga ngabo mu miyoboro y’amaraso ishobora guterwa n’umuvuduko uterwa no kuba hari intanga ngabo nyinshi mu mwanya muto.
Mu gusuzuma imyitwarire y’intanga ngabo ku birahuri bishya bimanitse, ibimenyetso by’ingufu z’intanga ngabo bishobora kugaragara, cyane cyane ku nkengero z’ibitonyanga by’intanga ngabo. Ariko, ingufu z’intanga ngabo zakunze kubangamira ibikorwa byo kuzenguruka bifitanye isano no kugenda ubudasiba, ibyo bikaba bisobanura imiterere y’igihe gito y’iki kintu. Abashakashatsi banabonye ko iyo inyunganizi yongewemo mu ntanga ngabo, hagaragaye utunyangingo turerire “dusa n’udutsi” twagaragaye.
Kugerageza kwa mbere kwigana intanga z'intanga byakozwe hakoreshejwe gukuraho insinga nto mu gitonyanga cyari cyamanitse, bituma intanga ndende isa n'intanga isohoka mu gitonyanga cy'intanga. Intanga z'intanga zahise zihuza mu buryo bumwe muri iyo ntanga, ariko igice cyose cyahise kibura bitewe n'aho 3D igarukira. Kubwibyo, kugira ngo wige uburyo intanga z'intanga ... Mero na Ogasawara14 bagabanyijemo utunyangingo tw’intanga ngabo mu tunyangingo twagutse n’utw’intanga ngabo tutanini, twombi twari dufite uburebure bumwe (~500 µm) n’ubugari bw’ijosi (~38 µm), ariko uburebure bw’impuzandengo y’utunyangingo bwari 56.6 na 56.6 µm. . , uko bikurikirana 11.2 μm. Mu bushakashatsi buriho ubu, twakoresheje igikoresho cya microfluidic gifite ingano ya 200 µm × 20 µm (W × H), igice cyacyo kiri hafi gato y’icya SST yazamuwe. Byongeye kandi, twasuzumye uburyo intanga zigenda n’imyitwarire yo gufungana mu mazi atemba, ibi bikaba bihuye n’igitekerezo cya Foreman cy’uko amazi akorwa n’utunyangingo twa epithelial twa SST atuma intanga ngabo ziguma mu kanyangingo mu cyerekezo kinyuranyije n’umuvuduko w’amaraso (rheological).
Intego y'ubu bushakashatsi kwari ukurwanya ibibazo byo kureba uko intanga zigenda mu muyoboro w'inkoko no kwirinda ingorane zo kwiga rheology n'imyitwarire y'intanga mu kirere gihindagurika. Hakoreshejwe igikoresho gitanga umuvuduko wa hydrostatic kugira ngo gishushanye uko intanga zigenda mu myanya ndangagitsina y'inkoko.
Iyo igitonyanga cy'intanga ngore yakuwemo amazi (1:40) cyashyirwaga mu gikoresho cya microchannel, hashoboraga kumenyekana ubwoko bubiri bw'uburyo intanga zigenda (intanga zitandukanijwe n'intanga zihambiriwe). Byongeye kandi, intanga ngore zakundaga koga zinyuranyije n'amazi (rheology nziza; videwo ya 1, 2). Nubwo intanga ngabo zari zifite umuvuduko muto ugereranyije n'intanga ngabo zonyine (p < 0.001), zongereye igipimo cy'intanga ngabo zigaragaza rheotaxis nziza (p < 0.001; Imbonerahamwe ya 2). Nubwo intanga ngabo zari zifite umuvuduko muto ugereranyije n'intanga ngabo zonyine (p < 0.001), zongereye igipimo cy'intanga ngabo zigaragaza rheotaxis nziza (p < 0.001; Imbonerahamwe ya 2). Хотя пучки спер тузоидов имели более низкую скорость, чем у одиночных спер мозозидов (p <0,001), Nubwo uduce tw’intanga ngabo twari dufite umuvuduko muto ugereranije n’uw’intanga ngabo imwe (p < 0.001), twongereye igipimo cy’intanga ngabo zigaragaza rheotaxis nziza (p < 0.001; Imbonerahamwe ya 2).尽管精子束的速度低于孤独精子的速度（ p <0.001 ），但它们增加了显示阳性流变性的精子百分比（ p <0.001 ；表 2 ）。尽管 精子束 的 速度 低于 孤独 的 （p <0.001） ， 但 增加 了 <<

Хотя скорость пу чов спер Музозоидов б ман ниже, чем у одиночных спер зозоидов (p <0,001), Nubwo umuvuduko w'intanga ngore wari munsi y'uwa spermatozoa imwe (p < 0.001), biyongereye igipimo cy'intanga ngore hamwe n'indwara ya rheology (p < 0.001; Imbonerahamwe ya 2).Impuzandengo nziza ku ngano imwe y'umura n'utubuto tw'umura bigereranywa kuri 53% na 85%, uko bikurikirana.
Byagaragaye ko intanga z'inkoko za sharkasi zikimara gusohora zikora uduce duto, tugizwe n'abantu benshi. Utwo duce twiyongera uburebure n'ubugari uko igihe kigenda kandi dushobora kuguma muri vitro amasaha menshi mbere yo gushwanyagurika (videwo ya 3). Utwo duce dufite uduce duto dufite ishusho nk'iy'intanga z'ingore zikora ku mpera ya epididymis. Byagaragaye ko intanga z'ingore za Sharkashi zifite ubushake bwo gushwanyagurika no gukora agace gafite imiyoboro mu gihe kitarenze umunota umwe nyuma yo gukusanywa. Utwo duce turahinduka kandi dushobora gufata ku nkuta zose zegereye cyangwa ibintu bidahinduka. Nubwo uduce duto tw'intanga tugabanya umuvuduko w'uturemangingo tw'intanga, biragaragara ko mu buryo bwa macroscopic twongera umurongo wazo. Uburebure bw'uduce buratandukanye bitewe n'umubare w'intanga zakusanyijwe mu duce duto. Ibice bibiri by'agace byashyizwe ukwabyo: igice cya mbere, harimo umutwe w'intanga washyizwe mu duce duto, n'igice cya nyuma, harimo umurizo n'iherezo ryose ry'intanga. Hakoreshejwe kamera yihuta cyane (950 fps), imitwe y’intanga ngabo yagaragaye mu gice cya mbere cy’urufunguzo, ikora ku buryo urufunguzo rugenda bitewe n’uko rugenda ruhindagurika, rukurura izindi mu rufunguzo mu buryo bwa helical (Videwo ya 4). Ariko, mu migozi miremire, byagaragaye ko imitwe imwe y’intanga ngabo yafashe ku mubiri kandi igice cya nyuma cy’urufunguzo gikora nk’ibice bifasha mu gusunika urufunguzo.
Mu gihe amazi agenda buhoro buhoro, uduce tw’intanga ngabo tugenda tubangikanye, ariko dutangira guhurirana no gufatana ku kintu cyose gihagaze, kugira ngo tudatwarwa n’amazi agenda uko umuvuduko w’amazi wiyongera. Utwo duce tubaho iyo uturemangingo tw’intanga ngabo duke twegeranye, dutangira kugenda mu buryo bw’ikigereranyo no gupfunyikana, hanyuma tugafatana ku kintu gifatana. Ishusho ya 1 na 2 igaragaza uburyo intanga ngabo zegerana, zigakora ihuriro mu gihe imirizo izingirana.
Abashakashatsi bakoresheje umuvuduko wa hydrostatic kugira ngo habeho gutembera kw'amazi mu muyoboro muto kugira ngo biga ibijyanye n'imikorere y'intanga ngore. Hakoreshejwe umuyoboro muto ufite ingano ya 200 µm × 20 µm (W × H) n'uburebure bwa 3.6 µm. Koresha imiyoboro mito hagati y'ibikoresho bifite inshinge ku mpera. Hakoreshejwe irangi ry'ibiryo kugira ngo imiyoboro igaragare neza.
Fata insinga n'ibindi bikoresho ku rukuta. Videwo yafashwe hakoreshejwe mikorosikopi y'icyiciro cyo kugereranya. Kuri buri shusho, hagaragazwa mikorosikopi y'icyiciro cyo kugereranya n'amashusho yo kugena aho ibintu bihurira. (A) Isano iri hagati y'imigezi ibiri irwanya amazi bitewe n'ingendo z'umuzunguruko (umwambi utukura). (B) Isano iri hagati y'umuyoboro w'amazi n'urukuta rw'umuyoboro (umwambi utukura), icyarimwe bihuzwa n'indi miyoboro ibiri (umwambi w'umuhondo). (C) Imiyoboro y'intanga ngabo iri mu muyoboro wa mikorosikopi itangira guhuzwa (umwambi utukura), igakora urusobe rw'imiyoboro y'intanga ngabo. (D) Ikorwa ry'urusobe rw'imiyoboro y'intanga ngabo.
Iyo igitonyanga cy'intanga ngore cyavangwaga cyashyirwaga mu gikoresho cy'amazi make maze hagakorwa urujya n'uruza, uruziga rw'intanga ngore rwagaragaye ko rugenda rugana aho urujya n'uruza rugana. Uduce duto dufata neza ku nkuta z'imiyoboro mito, kandi imitwe iri mu gice cya mbere cy'uduce duto iradufata neza (videwo ya 5). Nanone zifata ku duce twose duhagaze mu nzira yazo, nk'imyanda, kugira ngo zirinde gutwarwa n'amazi. Uko igihe kigenda gihita, utu duce duhinduka uduce turerure dufata izindi ntanga ngore imwe n'uduce tugufi (videwo ya 6). Uko urujya n'uruza rutangira kugenda buhoro, imirongo miremire y'intanga itangira gukora urusobe rw'imirongo y'intanga ngore (videwo ya 7; Ishusho ya 2).
Ku muvuduko mwinshi w’amazi (V > 33 µm/s), ingendo z’imigozi ziyongera mu rwego rwo gufata intanga nyinshi zikora uduce duto kugira ngo zirwanye imbaraga zo gutemba kw’amazi. Ku muvuduko mwinshi w’amazi (V > 33 µm/s), ingendo z’imigozi ziyongera mu rwego rwo gufata intanga nyinshi zikora uduce duto kugira ngo zirwanye imbaraga zo gutemba kw’amazi. При высокой скорости потока (V> 33 мкм / с) спиралевидные мжжения нитей усиливаются, поскольку они пытаются пой мать множвество отдельных спер язозоидч, пучю потока. Ku muvuduko mwinshi w’amazi (V > 33 µm/s), ingendo z’imigozi zigenda ziyongera uko zigerageza gufata intanga nyinshi zikora uduce duto dushobora kurwanya imbaraga zo gutemba kw’amazi.V (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漂移力。在 高 流速 (v> 33 µm / s) 时 ， 的 螺旋 运动 增加 ， 以 许多 许多 形成 单 个 精子 ， 漂移力。。。。。。。。。。 При высоких скоростях потока (V> 33 мкм / с) спиральное мжжение нитей увеличивается в попытке захитит множество отдельных спер позозоидов, обрующих пучки, ч кый лучше сопрочлч. Iyo amazi atemba cyane (V > 33 µm/s), urujya n'uruza rw'imitsi ruriyongera mu rwego rwo gufata intanga nyinshi zikora uduce kugira ngo zirwanye neza imbaraga z'amazi atemba.Bagerageje kandi gushyira imiyoboro mito ku nkuta z'impande.
Uduce tw’intanga ngabo twagaragaye nk’imitwe y’intanga ngabo n’imirizo izunguruka hakoreshejwe mikorosikopi yoroheje (LM). Uduce tw’intanga ngabo dufite uduce dutandukanye twagaragaye nk’imitwe izunguruka n’imitwe izunguruka, imirizo myinshi izunguruka, imitwe y’intanga ngabo ifatanye n’umurizo, n’imitwe y’intanga ngabo ifite nuclei izunguruka nk’imitwe myinshi izunguruka. Mikorosikopi y’amashanyarazi yo mu bwoko bwa scanning (SEM) yagaragaje ko uduce tw’intanga ngabo twari dufatanye mu gisenge cy’imitwe y’intanga ngabo kandi utwo duce tw’intanga ngabo twerekanye urusobe rw’imirizo izunguruka.
Imiterere n'imiterere ya spermatozoa, ikorwa ry'udutsiko tw'intanga byakorewe ubushakashatsi hakoreshejwe mikoroskopi yoroheje (igice cya kabiri cy'igice), mikoroskopi ya electron scanning (SEM) na mikoroskopi ya electron transmission (TEM), smears z'intanga zasizwe irangi ry'icunga rya acridine hanyuma zigasuzumwa hakoreshejwe mikoroskopi ya epifluorescence.
Ibara ry'intanga ngore rikoreshejwe akridine orange (Ishusho ya 3B) ryagaragaje ko imitwe y'intanga ngore yari ifatanye kandi itwikiriwe n'ibikoresho bisohora, byatumye habaho uduce duto (Ishusho ya 3D). Uduce tw'intanga ngore twari tugizwe n'uduce tw'intanga ngore dufite urusobe rw'imirizo ifatanye (Ishusho ya 4A-C). Uduce tw'intanga ngore tugizwe n'imirizo y'intanga ngore nyinshi zifatanye (Ishusho ya 4D). Amabanga (Ishusho ya 4E, F) yari atwikirije imitwe y'uduce tw'intanga ngore.
Imiterere y'uruvange rw'intanga ngabo Hakoreshejwe mikorosikope y'ibara ry'ikinyuranyo cy'ibara n'imisemburo y'intanga ngabo yasizweho irangi ry'icunga rya akridine, byagaragaje ko imitwe y'intanga ngabo ifatana. (A) Imiterere y'intanga ngabo itangirana n'intanga ngabo (uruziga rw'umweru) n'intanga ngabo eshatu (uruziga rw'umuhondo), aho uruziga ruhera ku murizo rukarangira ku mutwe. (B) Fotokopi y'uruvange rw'intanga ngabo yasizweho irangi ry'icunga rya akridine igaragaza imitwe y'intanga ngabo ifatana (imyambi). Amasohoro atwikira umutwe (imitwe). Ubwinshi × 1000. (C) Iterambere ry'uruvange runini rutwarwa n'amazi mu muyoboro wa microfluidic (hakoreshejwe kamera yihuta cyane kuri 950 fps). (D) Mikorokopi y'uruvange rw'intanga ngabo yasizweho irangi ry'icunga rya akridine igaragaza imitumba minini (imyambi). Ubwinshi: × 200.
Gupima mikorografi ya elegitoroni y'umurabyo w'intanga n'intanga zishushanyijeho ibara ry'icunga rya akridine. (A, B, D, E) ni mikorografi ya elegitoroni y'intanga zishushanyijeho ibara ry'ikoranabuhanga, naho C na F ni mikorografi ya mikorografi ya akridine y'icunga igaragaza ko intanga nyinshi zipfundikiye urusobe rw'intanga. (AC) Intanga zishyirwa hamwe zigaragara nk'urusobe rw'imirizo ifatanye (imyambi). (D) Gufata intanga nyinshi (zifite ikintu gifata, ishusho y'umutuku, umwambi) zizunguruka umurizo. (E na F) Intanga zishyirwa hamwe (icyerekezo) zitwikiriwe n'ibikoresho bifatanye (icyerekezo). Intanga zikora uduce dufite imiterere myinshi isa n'iy'urumuri (F). (C) ×400 na (F) ×200.
Dukoresheje mikorosikope ya elegitoroniki yo kohereza, twasanze uduce tw’intanga ngabo dufite imirizo ifatanye (Ishusho ya 6A, C), imitwe ifatanye n’imirizo (Ishusho ya 6B), cyangwa imitwe ifatanye n’imirizo (Ishusho ya 6D). Imitwe y’intanga ngabo iri mu gasanduku iragoramye, igaragaza mu gice cya kabiri cy’uturere twa nuclear (Ishusho ya 6D). Mu gasanduku k’isuka, intanga ngabo yari ifite umutwe ugoramye ufite uduce tubiri twa nuclear n’uduce twinshi twa flagellum (Ishusho ya 5A).
Mikorografi y'ikoranabuhanga yerekana imirizo ihuza mu gipfunyika cy'intanga n'ibikoresho bihuza imitwe y'intanga. (A) Umurizo ufatanye w'umubare munini w'intanga. Reba uko umurizo ugaragara haba mu ishusho (umwambi) no mu miterere y'ubutaka (umwambi). (B) Umutwe (umwambi) w'intanga ufatanye n'umurizo (umwambi). (C) Imirizo myinshi y'intanga (imyambi) ifatanye. (D) Ibikoresho byo guhuza (AS, ubururu) bihuza imitwe ine y'intanga (umuhengeri).
Mikroskopi ya elegitoroniki yo gupima yakoreshejwe mu kumenya imitwe y'intanga mu dupfunyika tw'intanga cyangwa utwenda (Ishusho ya 6B), bigaragaza ko udupfunyika tw'intanga twari dufatanye n'ibintu byo hanze y'inyuma. Ibikoresho byashyizwe mu mutwe w'intanga byashyizwe mu mutwe w'intanga (umutwe wa jellyfish umeze nk'umutwe; Ishusho ya 5B) kandi byagutse cyane, bitanga isura y'umuhondo mwiza munsi ya mikroskopi ya fluorescence iyo yasizweho akridine orange (Ishusho ya 6C). Iki kintu kigaragara neza munsi ya mikroskopi yo gupima kandi gifatwa nk'ikintu gifata. Ibice bito cyane (Ishusho ya 5C) n'imisemburo y'intanga byasizweho akridine orange byagaragaje udupfunyika tw'intanga turimo imitwe myinshi n'imirizo igonze (Ishusho ya 5D).
Fotografi zitandukanye zigaragaza ukwishyira hamwe kw'imitwe y'intanga n'imirizo ipfundikiye hakoreshejwe uburyo butandukanye. (A) Mikorografi y'ikoranabuhanga yo kohereza amabara ya elegitoroniki y'umugozi w'intanga igaragaza umutwe w'intanga upfundikiye ufite igice kibiri (ubururu) n'ibice byinshi by'ibendera (icyatsi). (B) Mikorografi y'ikoranabuhanga yo gupima amabara ya elegitoroniki yerekana itsinda ry'imitwe y'intanga isa n'iy'ifi isa n'ipfundikiye. (C) Igice gito cyane kigaragaza imitwe y'intanga ipfundikiye (imyambi) n'imirizo ipfundikiye (imyambi). (D) Mikorografi y'intanga ipfundikiye ibara ry'icunga rya acridine igaragaza imitwe y'intanga ipfundikiye (imyambi) n'imirizo ipfundikiye (imyambi). Menya ko ikintu gifata (S) gitwikira umutwe w'intanga. (D) × 1000 magnification.
Hakoreshejwe mikoroskopi ya elegitoroniki yo kohereza (Ishusho ya 7A), byagaragaye kandi ko imitwe y'intanga yari yazungurutse kandi ko ingirangingo zayo zari zifite ishusho y'uruziga, nk'uko byemejwe n'imisemburo y'intanga yasizwe ibara ry'icunga rya akridine kandi igasuzumwa hakoreshejwe mikoroskopi ya fluorescence (Ishusho ya 7B).
(A) Mikorografi ya elegitoroniki yo kohereza amabara mu buryo bwa digitale na (B) Isuzuma ry'intanga ngore rya Acridine orange rigaragaza imitwe izungurutse n'aho imitwe n'imirizo by'intanga ngore bifatanye (imyambi). (B) × 1000 magnification.
Igishimishije ni uko intanga za Sharkazi ziterana zigakora uduce tw’imigozi tugenda. Imiterere y’utu duce idufasha gusobanukirwa uruhare rushoboka rwatwo mu kwinjiza no kubika intanga muri SST.
Nyuma yo guhuza, intanga zinjira mu gitsina zigahita zitoranywa cyane, bigatuma umubare muto w’intanga winjira muri SST15,16. Kugeza ubu, uburyo intanga zinjira zigasohoka muri SST ntiburasobanuka neza. Mu nkoko, intanga zibikwa muri SST mu gihe cy’ibyumweru 2 kugeza ku 10, bitewe n’ubwoko6. Haracyari impaka ku miterere y’intanga mu gihe zibikwa muri SST. Ese ziri mu nzira cyangwa ziruhutse? Mu yandi magambo, ni gute intanga zigumana aho zihagaze muri SST igihe kirekire?
Forman4 yavuze ko aho intanga zituye n'aho zisohoka bishobora gusobanurwa mu buryo intanga zigenda. Abanditsi bavuga ko intanga zigumana aho zihagaze zoga zigana ku mazi atemba aturuka kuri epithelium ya SST kandi ko intanga zisohoka muri SST iyo umuvuduko wazo ugabanutse munsi y'aho zitangiye gusubira inyuma kubera kubura imbaraga. Zaniboni5 yemeje ko hari aquaporins 2, 3, na 9 mu gice cy'uturemangingo twa SST, bishobora gushyigikira uburyo bwo kubika intanga bwa Foreman. Mu bushakashatsi bwakozwe ubu, twasanze hafi kimwe cya kabiri cy'intanga za Sharkashi zigaragaza rheology nziza mu mazi atemba, kandi ko intanga zivanze zongera umubare w'intanga zigaragaza rheology nziza, nubwo agglutination izitinza umuvuduko. Uburyo intanga zigenda mu muyoboro w'inkoko zijya aho intanga zishyingirwa ntabwo byumvikana neza. Mu nyamaswa, amenyo ya follicular fluid chemotherapy ikurura intanga. Ariko, imiti ikurura intanga ikekwaho kuyobora intanga kwegera intera ndende7. Kubwibyo, hari izindi nzira zituma intanga zitwara. Ubushobozi bw'intanga bwo kwerekeza no gutembera ku muyoboro w'amazi usohoka nyuma yo gukururana byavuzwe ko ari ikintu gikomeye mu kwibasira intanga mu mbeba. Parker 17 yavuze ko intanga zinyura mu mabya zikoga zigana ku muyoboro w'amazi mu nyoni no mu bikurura. Nubwo bitaragaragazwa mu buryo bw'igerageza mu nyoni, Adolphi18 ni we wa mbere wasanze intanga ziguruka zitanga umusaruro mwiza iyo urwego ruto rw'amazi hagati y'urupapuro rwometseho n'urushundura rukozwe n'agapapuro k'urushundura. Rheology. Hino na Yanagimachi [19] bashyize ingano y'intanga-utubari-nyababyeyi mu ruziga rw'amazi maze batera µl 1 y'inyi mu gice cy'amazi kugira ngo babone uko amazi atembera mu miyoboro y'amazi. Babonye ko hari urujya n'uruza rwinshi mu muyoboro w'amazi, aho udupira twose tw'inyinyo twagendaga tugana ku ruhu rw'umuyoboro w'amazi. Abanditsi bashimangira akamaro k'amazi y'imiyoboro y'amazi ava hasi ajya ku muyoboro wo hejuru w'amazi kugira ngo intanga zizamuke kandi zigere ku nda. Brillard20 yavuze ko mu nkoko n'ingurube, intanga zigenda zimuka zivuye mu muryango w'igitsina gore, aho zibikwa, zijya mu ihuriro ry'umura n'igitsina gore, aho zibikwa. Ariko, uku kugenda ntigusabwa hagati y'ihuriro ry'umura n'infundibulum kuko intanga zitwarwa no kwimuka. Tumenye izi nama zabanje n'ibyavuye mu bushakashatsi buriho, dushobora gutekereza ko ubushobozi bw'intanga zigenda zizamuka (rheology) ari kimwe mu bintu inzira yo guhitamo ishingiyeho. Ibi bigena inzira intanga zinyuramo mu gitsina ndetse n'uburyo zinjira muri CCT kugira ngo zibikwe. Nk'uko Forman4 yabivuze, ibi bishobora no koroshya inzira intanga zinjira muri SST n'aho ziba mu gihe runaka hanyuma zigasohoka iyo umuvuduko wazo utangiye kugabanuka.
Ku rundi ruhande, Matsuzaki na Sasanami 21 bavuze ko intanga ngabo zihinduka mu mikorere yazo kuva ku kurara kugeza ku kumererwa neza mu myanya myibarukiro y’abagabo n’abagore. Hagaragajwe ko intanga ngabo zitagira imikorere myiza mu mikorere yazo kugira ngo hasobanurwe igihe kirekire intanga ngabo zibikwamo hanyuma zikongera kumererwa neza nyuma yo kuva muri SST. Mu gihe cy’ubushyuhe bwinshi, Matsuzaki n’abandi bavuze ko umusaruro mwinshi wa lactate muri SST, bishobora gutuma intanga ngabo zitagira imikorere myiza. Muri iki gihe, akamaro k’intanga ngabo kagaragarira mu guhitamo no kwinjiza intanga ngabo, atari mu kubika kwazo.
Imiterere y’intanga ngore ifatwa nk'ibisobanuro byumvikana ku gihe kirekire intanga ngore zibikwa muri SST, kuko iyi ari ihame risanzwe ryo kugumana intanga mu nkoko2,22,23. Bakst n'abandi 2 babonye ko intanga nyinshi zafatanaga, zigakora uduce duto, kandi intanga imwe ntiyakunze kuboneka muri CCM y'inkware. Ku rundi ruhande, Wen n'abandi 24 babonye intanga nyinshi zitatanye kandi intanga nke zigasigara mu ruhu rwa SST mu nkoko. Dukurikije ibi byagaragaye, dushobora gutekereza ko ubushake bwo guteranya intanga butandukanye hagati y'inyoni n'intanga mu ruhu rumwe. Byongeye kandi, Van Krey n'abandi 9 bavuze ko kwitandukanya kw'intanga ngore mu buryo butunguranye ari byo bituma intanga zinjira buhoro buhoro mu ruhu rw'umuyoboro w'inda. Dukurikije iki gitekerezo, intanga ngore zifite ubushobozi buke bwo guteranya intanga zigomba kubanza gukurwa muri SST. Muri uru rwego, ubushobozi bw'intanga ngabo bwo kwihuza bushobora kuba ikintu bigira ingaruka ku musaruro w'irushanwa ry'intanga ngabo mu nyoni zanduye. Byongeye kandi, uko intanga ngabo zihuza igihe kirekire, niko uburumbuke bukomeza kuba bwinshi.
Nubwo kwishyira hamwe kw'intanga no kwishyira hamwe mu matsinda byagaragaye mu bushakashatsi butandukanye2,22,24, ariko ntibyasobanuwe mu buryo burambuye bitewe n'uburyo zigaragara mu buryo bugoye muri SST. Hakozwe ubushakashatsi bwinshi bwo kwiga kwishyira hamwe kw'intanga mu buryo bwa vitro. Kwishyira hamwe gukabije ariko by'igihe gito byagaragaye ubwo insinga nto yakurwaga ku gitonyanga cy'imbuto cyari kimanitse. Ibi bituma agahu kanini gasohoka mu gitonyanga, gakigana igitsina cy'intanga. Bitewe n'ubushobozi bwo kugabanya imiterere ya 3D no kuma igihe gito cy'amazi, agace kose karangiritse vuba9. Mu bushakashatsi buriho, dukoresheje inkoko za Sharkashi n'udusimba twa microfluidic, twashoboye gusobanura uburyo utu dusimba dukora n'uko twimuka. Udusimba tw'intanga twakozwe ako kanya nyuma yo gukusanya amasohoro kandi byagaragaye ko twimuka mu buryo buziguye, bigaragaza rheology nziza iyo turi mu ruziga. Byongeye kandi, iyo harebwe mu buryo bwa macroscopic, utwo dusimba tw'intanga twagaragaye ko twongera umurongo w'imikorere ugereranije n'udusimba twihariye. Ibi byerekana ko guterana kw'intanga bishobora kubaho mbere yuko SST yinjira kandi ko umusaruro w'intanga utagarukira ku gace gato bitewe n'umuvuduko nk'uko byavuzwe mbere (Tingari na Lake12). Mu gihe cyo gukora umuyoboro w'intanga, intanga ziroga mu buryo buhuye kugeza igihe zikoze aho zihurira, hanyuma imirizo yazo irazengurukana maze umutwe w'intanga zigakomeza kuba zidafite aho zibogamiye, ariko umurizo n'igice cyo hagati cy'intanga zifatana hamwe n'ikintu gifatana. Kubwibyo, umutwe w'intanga zifatana ni wo utuma bigenda, ukurura igice gisigaye cy'intanga. Gupima mikorosikope y'intanga zifatana byagaragaje imitwe y'intanga zifatana itwikiriwe n'ibintu byinshi bifatana, bigaragaza ko imitwe y'intanga yari ifatanye mu dupfunyika, ibyo bikaba byarabaye nyuma yo kugera aho ububiko bubikwa (SST).
Iyo intanga ngabo zikoze ibara ry’umuhondo wa akridine, ibikoresho byo mu bwoko bwa kole bikikije intanga ngabo bishobora kugaragara hifashishijwe mikorosikopi y’urumuri. Iyi ngirabuzimafatizo yemerera intanga ngabo gufata no gufata ku buso cyangwa uduce duto dukikije kugira ngo tudatembera hamwe n’amazi akikije intanga. Bityo, ibyo twabonye bigaragaza uruhare rw’intanga ngabo mu buryo bw’uduce duto. Ubushobozi bwazo bwo koga zihanganye n’amazi no gufata ku buso bwegereye butuma intanga ngabo ziguma igihe kirekire muri SST.
Rothschild25 yakoresheje kamera ya hemocytometry kugira ngo yige uburyo amasohoro y’inka akwirakwira mu gitonyanga cy’urumuri, afata fotomikorografi binyuze muri kamera ifite umurongo ugororotse n’ugororotse wa mikorosikopi. Ibisubizo byagaragaje ko intanga ngabo zakururwaga n’ubuso bw’icyumba. Abanditsi bavuga ko hashobora kubaho imikoranire ya hydrodynamic hagati y’intanga ngabo n’ubuso. Dukurikije ibi, hamwe n’ubushobozi bw’intanga ngabo za Sharkashi bwo gukora uduce duto, bishobora kongera amahirwe y’uko amasohoro azaguma ku rukuta rwa SST kandi akabikwa igihe kirekire.
Bccetti na Afzeliu26 bavuze ko glycocalyx y'intanga ikenewe kugira ngo hamenyekane gamete kandi hakorwe agglutination. Forman10 yabonye ko hydrolysis y'imigozi ya α-glycosidic mu migozi ya glycoprotein-glycolipid binyuze mu kuvura intanga z'inkoko na neuraminidase byatumye uburumbuke bugabanuka nta ngaruka ku ihinduka ry'intanga. Abanditsi bavuga ko ingaruka za neuraminidase kuri glycocalyx zibangamira uburyo intanga zishyirwa mu ihuriro ry'intanga n'abagore, bityo zikagabanya uburumbuke. Ibyo babonye ntibishobora kwirengagiza ko uburyo bwo kuvura neuraminidase bushobora kugabanya imenyekana ry'intanga n'uruhinja. Forman na Engel10 basanze uburumbuke bwaragabanutse igihe inkoko zaterwaga intanga mu gitsina zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigaterwa intanga zigafatwa ... Abanditsi banzuye ko impinduka mu gishishwa cya glycoprotein-glycolipide gikikije ururenda rw'intanga ngore zagabanyije ubushobozi bw'intanga ngore bwo gukurura intanga ngore binyuze mu kwangiza uburyo intanga ngore zihuzwa n'ururenda rw'intanga ngore, ibyo bikaba byaratumye intanga ngore zitakaza umuvuduko bitewe n'umuvuduko w'ururenda rw'intanga ngore, ariko ntibigire ingaruka ku kumenyekana kw'intanga ngore n'igi.
Mu biguruka, Bakst na Bauchan 11 basanze uduce duto tw’uturemangingo n’uduce tw’uturemangingo mu ruhu rw’intanga ngabo, maze babona ko bimwe muri ibyo binyabutabire byari byarahujwe n’uruhererekane rw’intanga ngabo. Abanditsi bavuga ko iyi mibanire ishobora kugira uruhare mu kubika intanga ngabo mu gihe kirekire. Ariko, abashakashatsi ntibagaragaje aho izi ngingo zikomoka, niba zituruka mu turemangingo twa CCT, zikorwa kandi zigakurwa mu buryo bw’imyororokere bw’abagabo, cyangwa se zikorwa n’intanga ngabo ubwayo. Nanone, izi ngingo nizo zituma intanga ngabo zikomeza kwiyongera. Grützner n’abandi27 bavuze ko uturemangingo twa epididymal dukora kandi tugakura poroteyine runaka ikenewe kugira ngo habeho uburyo bwo gukora igice kimwe cy’imisemburo. Abanditsi banavuga ko gukwirakwira kw’izi ngingo biterwa n’imikoranire ya poroteyine za epididymal. Nixon n’abandi28 basanze adnexa ikora poroteyine, osteonectin ikungahaye kuri cysteine; SPARC igira uruhare mu ikorwa ry’uduce tw’intanga ngabo mu turemangingo tugufi twa echidnas na platypuses. Gukwirakwira kw'iyi mirasire bifitanye isano no kubura iyi poroteyine.
Mu bushakashatsi bwakozwe ubu, isesengura ry’imiterere y’inyubako hakoreshejwe mikorosikopi ya elegitoroniki ryagaragaje ko intanga ngabo zifata ku bwinshi bw’ibintu byinshi byuzuye. Ibi bintu bivugwa ko ari byo bituma intanga ngabo zifatana hagati no hafi y’imitwe ifatana, ariko ku rugero ruto mu gace k’umurizo. Dutekereza ko iki kintu gifatana gisohoka mu buryo bw’imyororokere bw’abagabo (epididymis cyangwa vas deferens) hamwe n’amasohoro, kuko akenshi tubona amasohoro atandukanye n’amaraso ya lymph na plasma y’intanga ngabo mu gihe cyo gusohora. Byavuzwe ko uko intanga ngabo z’inyoni zinyura muri epididymis na vas deferens, zihura n’impinduka zijyanye no gukura zishyigikira ubushobozi bwazo bwo gufata poroteyine no kubona glycoproteine ​​zifitanye isano na plasma lemma. Kuba izi poroteyine ziguma ku mpande z’intanga ngabo muri SST bigaragaza ko izi poroteyine zishobora kugira ingaruka ku buryo intanga ngabo zigumana neza 30 kandi zikagena uburumbuke bwazo 31. Ahammad n'abandi32 bavuze ko intanga ngabo zikomoka mu bice bitandukanye by'imyanya myibarukiro y'abagabo (kuva ku mabya kugeza ku gice cy'inyuma cy'uturemangingo tw'amaraso) zagaragaje ko zikomeza kwiyongera mu mikorere y'amazi mu gihe cyo kubika, hatitawe ku bushyuhe bwo kubika, kandi ko imikoreshereze y'inkoko nayo yiyongera mu miyoboro y'inda nyuma yo gutera intanga mu buryo bw'ubukorano.
Udukoko tw’inkoko twa Sharkashi dufite imiterere n’imikorere itandukanye n’andi moko nka echidnas, platypus, imbeba zo mu giti, imbeba z’impala, n’ingurube zo mu bwoko bwa Guinea. Mu nkoko za sharkasi, ikorwa ry’udukoko tw’intanga ryagabanije umuvuduko wazo wo koga ugereranije n’udukoko tw’intanga imwe. Ariko, utu dukoko twongereye igipimo cy’udukoko twa rheologically kandi twongera ubushobozi bw’udukoko bwo kwihagararaho mu buryo buhindagurika. Bityo, ibisubizo byacu byemeza igitekerezo cyabanje cy’uko guterana kw’intanga muri SST bifitanye isano no kubika intanga igihe kirekire. Nanone dutekereza ko gukunda gukora udukoko bishobora kugenzura umuvuduko w’ibura ry’intanga muri SST, bishobora guhindura umusaruro w’irushanwa ry’intanga. Dukurikije iki gitekerezo, spermatozoa ifite ubushobozi buke bwo guterana intanga ibanza kurekura SST, mu gihe spermatozoa ifite ubushobozi bwinshi bwo guterana intanga ibyara abana benshi. Ikorwa ry’udukoko tw’intanga imwe ni ingirakamaro kandi rigira ingaruka ku gipimo cy’umubyeyi n’umwana, ariko rikoresha uburyo butandukanye. Muri echidnas na platypus, intanga zitondetse zifatanye kugira ngo zongere umuvuduko w'imbere w'urumuri. Imigozi ya echidnas igenda vuba inshuro eshatu kurusha intanga imwe. Bikekwa ko kuremwa kw'intanga nk'izo muri echidnas ari uburyo bwo kwiyubaka kugira ngo zikomeze kwiganza, kubera ko abagore baba ari ubusambanyi kandi akenshi babana n'abagabo benshi. Kubwibyo, intanga zikomoka ku misemburo itandukanye zirushanwa cyane kugira ngo igi ribeho.
Intanga ngabo z’inkoko za sharkasi zoroshye kuzireba hakoreshejwe mikorosikopi y’ibara ritandukanye, ibi bifatwa nk'ingirakamaro kuko bituma byoroha kwiga imyitwarire y’intanga ngabo muri vitro. Uburyo intanga ngabo zikora butuma inkoko zororoka mu nkoko za sharkasi butandukana n’ubugaragara mu nyamaswa zimwe na zimwe zo mu nyababyeyi zigaragaza imyitwarire y’intanga ngabo nk’imbeba zo mu giti, aho zimwe mu ntanga ngabo zigera ku magi, bigafasha izindi mpungenge kugera no kwangiza amagi yazo. kugira ngo wigaragaze. imyitwarire y’ubwitange. Kwishyira mu mwanya w’abandi 34. Urundi rugero rw’imyitwarire y’ubufatanye mu ntanga ngabo rwabonetse mu mbeba z’impala, aho intanga ngabo zashoboye kumenya no guhuriza hamwe n’intanga ngabo zifitanye isano cyane n’uturemangingo no gukora amatsinda y’ubufatanye kugira ngo zongere umuvuduko wazo ugereranije n’intanga ngabo zitafitanye isano35.
Ibisubizo byabonetse muri ubu bushakashatsi ntibivuguruza inyigisho ya Foman yo kubika intanga mu gihe kirekire muri SWS. Abashakashatsi bavuga ko intanga zikomeza kugenda mu rujya n'uruza rw'uturemangingo tw’imisemburo dukikije SST mu gihe kirekire, kandi nyuma y'igihe runaka, ingufu zibikwa mu ngirabuzimafatizo z'intanga ziragabanuka, bigatuma umuvuduko ugabanuka, bigatuma imbaraga z'intanga zisohoka mu ruhu rw'amazi aturuka mu ruhu rw'imisemburo ya SST. Muri ubu bushakashatsi, twabonye ko kimwe cya kabiri cy'intanga imwe cyagaragaje ubushobozi bwo koga zihanganye n'amazi atemba, kandi gufatana kwazo mu gipfunyika byongera ubushobozi bwazo bwo kugaragaza rheology nziza. Byongeye kandi, amakuru yacu arahuye n'aya Matsuzaki n'abandi 1 bavuze ko kwiyongera kwa lactate muri SST bishobora kubuza intanga zituye kugenda. Ariko, ibisubizo byacu bivuga ku iremwa ry'imitsi igenda y'intanga n'imyitwarire yazo ya rheological mu gihe hari ahantu hahindagurika mu muyoboro muto mu rwego rwo gusobanura imyitwarire yazo muri SST. Ubushakashatsi bw'ejo hazaza bushobora kwibanda ku kumenya imiterere y'ibinyabutabire n'inkomoko y'ikintu gitera ubwiyongere bw'intanga ngabo, nta gushidikanya ko bizafasha abashakashatsi gutegura uburyo bushya bwo kubika amasohoro y'amazi no kongera igihe cy'uburumbuke.
Inyoni 15 z'igitsina gabo zifite ijosi ritagira ijosi (Na Na) zifite ibyumweru 30 zatoranijwe nk'abatanga intanga mu bushakashatsi. Izi nyoni zororewe mu ifumbire y'inkoko y'ishami ry'ubuhinzi, Kaminuza ya Ashit, Intara ya Ashit, mu Misiri. Inyoni zashyizwe mu tuzu dutandukanye (30 x 40 x 40 cm), zikorerwa gahunda y'urumuri (amasaha 16 y'urumuri n'amasaha 8 y'umwijima) kandi zigaburirwa indyo irimo garama 160 za poroteyine, 2800 kcal z'ingufu zishobora gukoreshwa mu gusya, garama 35 za kalisiyumu buri imwe. Garama 5 za fosifore iboneka kuri kilogarama y'indyo.
Dukurikije amakuru ya 36, ​​​​37, amasohoro yakuwe mu bagabo hakoreshejwe massage yo mu nda. Ingero 45 z'amasohoro zakuwe mu bagabo 15 mu minsi 3. Intanga ngabo (n = 15 ku munsi) zahise zivangwa muri 1:1 (v:v) hamwe na Belsville Poultry Semen Diluent, irimo potasiyumu diphosphate (1.27 g), monosodium glutamate monohydrate (0.867 g), fructose (0.5 d) sodium anhydrous. acetate (0.43 g), tris (hydroxymethyl)aminomethane (0.195 g), potasiyumu citrate monohydrate (0.064 g), potasiyumu monophosphate (0.065 g), magnesium chloride (0.034 g) na H2O (100 ml), pH = 7, 5, osmolarity 333 mOsm/kg38. Ingero z'amasohoro yakuwemo amazi zabanje gusuzumwa hifashishijwe mikorosikopi yoroheje kugira ngo harebwe ko amasohoro ari meza (ubushuhe) hanyuma zikabikwa mu bwogero bw'amazi kuri dogere selisiyusi 37 kugeza igihe zikoreshejwe mu gihe kitarenze iminota 30 nyuma yo kuyafata.
Isesengura ry’imiterere y’intanga ngore n’imiterere y’intanga bisobanurwa hakoreshejwe uburyo bw’ibikoresho bito bito. Ingero z’intanga ngore zongerewe ku gipimo cya 1:40 muri Beltsville Avian Semen Diluent, zishyirwa mu gikoresho gito bito (reba hepfo), kandi ibipimo bya kinetic byagenwe hakoreshejwe uburyo bwa Computerized Semen Analysis (CASA) bwakozwe mbere kugira ngo bugaragaze microfluidics. ku buryo intanga ngore zigenda mu buryo bw’amazi (Ishami ry’Ubwubatsi bw’Imashini, Ishami ry’Ubwubatsi, Kaminuza ya Assiut, Misiri). Iyi plugin irashobora gukururwa kuri: http://www.assiutmicrofluidics.com/research/casa39. Umuvuduko w’inyuguti (VCL, μm/s), umuvuduko w’umurongo (VSL, μm/s) n’umuvuduko w’umurongo (VAP, μm/s) byarapimwe. Amashusho y’intanga ngore yafashwe hakoreshejwe mikorosikopi ya Optika XDS-3 phase contrast (ifite intego ya 40x) ihujwe na kamera ya Tucson ISH1000 kuri 30 fps mu gihe cy’amasegonda 3. Koresha porogaramu ya CASA kugira ngo wige nibura ibice bitatu n'inzira 500 z'intanga ngore kuri buri gipimo. Videwo yafashwe hakoreshejwe CASA yakorewe mu rugo. Ibisobanuro by'uburyo bwo kugenda muri CASA bishingiye ku muvuduko wo koga kw'intanga ugereranije n'umuvuduko w'amazi, kandi ntibikubiyemo ibindi bipimo nko kugenda uruhande rumwe, kuko byagaragaye ko ari byiza mu gutembera kw'amazi. Uburyo bwo kugenda kw'intanga ngore busobanurwa nk'ingendo z'uturemangingo tw'intanga ngore tunyura mu cyerekezo cy'amazi atembera. Intanga ngore ifite imiterere ya rheological yagabanyijwemo umubare w'intanga ngore zigenda; intanga ngore zari zihagaze kandi zigenda zihuza n'intanga ngore ntizashyizwe mu mubare.
Imiti yose yakoreshejwe yakuwe muri Elgomhoria Pharmaceuticals (Cairo, Misiri) keretse iyo byavuzwe ukundi. Iki gikoresho cyakozwe nk'uko byasobanuwe na El-sherry et al. 40 hamwe n'impinduka zimwe na zimwe. Ibikoresho byakoreshejwe mu gukora imiyoboro mito birimo plaque z'ikirahure (Howard Glass, Worcester, MA), SU-8-25 negative resist (MicroChem, Newton, CA), diacetone alcohol (Sigma Aldrich, Steinheim, Ubudage), na polyacetone. -184, Dow Corning, Midland, Michigan). Imiyoboro mito ikorwa hakoreshejwe lithography yoroshye. Ubwa mbere, agapfukamunwa gafite imiterere ya microchannel kifuzwaga kacapishijwe kuri printer ifite ubushobozi bwo hejuru (Prismatic, Cairo, Misiri na Pacific Arts and Design, Markham, ON). Ibikoresho byakozwe hakoreshejwe plaque z'ikirahure nk'ibikoresho. Amasahani yogejwe muri acetone, isopropanol n'amazi yakuwemo iion hanyuma ashyirwa ku urwego rwa 20 µm rwa SU8-25 hakoreshejwe spin coating (3000 rpm, umunota 1). Hanyuma ibice bya SU-8 byumye buhoro buhoro (65°C, iminota 2 na 95°C, iminota 10) hanyuma bishyirwa ku mirasire ya UV mu gihe cy'amasegonda 50. Nyuma yo guteka kuri 65°C na 95°C mu gihe cy'umunota 1 n'iminota 4 kugira ngo ibice bya SU-8 bigaragare, hagakurikiraho guteranya alcohol ya diacetone mu gihe cy'iminota 6.5. Teka cyane waffles (200°C mu gihe cy'iminota 15) kugira ngo urwego rwa SU-8 rukomeze gukomera.
PDMS yateguwe hakoreshejwe kuvanga monomer na hardener mu gipimo cy'uburemere cya 10:1, hanyuma ikurwamo imyuka mu cyuma gisukura umwuka hanyuma isukwa ku gitereko cy'ingenzi cya SU-8. PDMS yavuwe mu ifuru (120°C, iminota 30), hanyuma imiyoboro iracibwa, itandukanywa n'icyuma gikurura, hanyuma iratoborwa kugira ngo imiyoboro ifatanye n'aho imiyoboro yinjira n'aho isohoka. Amaherezo, imiyoboro mito ya PDMS yashyizwe burundu kuri mikorosikopi hakoreshejwe poroseseri ya corona (Electro-Technic Products, Chicago, IL) nk'uko byasobanuwe ahandi. Umuyoboro mito wakoreshejwe muri ubu bushakashatsi upima 200 µm × 20 µm (W × H) kandi ufite uburebure bwa cm 3.6.
Urujya n'uruza rw'amazi ruterwa n'umuvuduko wa hydrostatic uri mu muyoboro muto rugerwaho binyuze mu kugumana urugero rw'amazi mu kigega cy'amazi cyinjira hejuru y'itandukaniro ry'uburebure Δh39 mu kigega cy'amazi gisohoka (Ishusho ya 1).
aho f ari coefficient of friction, isobanurwa nka f = C/Re ku bijyanye n'umuvuduko wa laminar mu muyoboro wa mpandeshatu, aho C ari constant bitewe n'ikigereranyo cy'umuyoboro, L ni uburebure bw'umuyoboro wa microchannel, Vav ni umuvuduko mpuzandengo uri imbere muri microchannel, Dh ni diameter ya hydraulic y'umuyoboro, g - kwihutisha imbaraga rukuruzi. Hakoreshejwe iki kigereranyo, umuvuduko mpuzandengo w'umuyoboro ushobora kubarwa hakoreshejwe iki kigereranyo gikurikira: