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Haihuwar tsuntsaye ya dogara ne da ikonsu na adana isasshen maniyyi mai rai na tsawon lokaci a cikin bututun ajiyar maniyyi (SST). Tsarin da maniyyi ke shiga, zama a ciki, da kuma barin SST ya ci gaba da zama abin muhawara. Manin kaji sharkasi ya nuna babban yanayin haɗuwa, yana samar da tarin filamentous masu motsi waɗanda ke ɗauke da ƙwayoyin halitta da yawa. Saboda wahalar lura da motsi da halayen maniyyi a cikin bututun fallopian mara haske, mun yi amfani da na'urar microfluidic tare da sashin giciye na microchannel mai kama da na maniyyi don nazarin haɗuwa da motsi na maniyyi. Wannan binciken ya tattauna yadda tarin maniyyi ke samuwa, yadda suke motsawa, da kuma yuwuwar rawar da suke takawa wajen faɗaɗa zama a cikin maniyyi a cikin SST. Mun binciki saurin maniyyi da halayen rheological lokacin da aka samar da kwararar ruwa a cikin tashar microfluidic ta hanyar matsin lamba na hydrostatic (yawan kwarara = 33 µm/s). Manya na iya yin iyo a kan yanayin da ake ciki (positive rheology) kuma saurin manya yana raguwa sosai idan aka kwatanta da manya guda ɗaya. An lura da tarin maniyyi suna motsawa a cikin karkace kuma suna ƙaruwa da tsayi da kauri yayin da ake samun ƙarin maniyyi guda ɗaya. An lura da tarin maniyyi suna kusantowa da mannewa a gefen hanyoyin microfluidic don gujewa sharewa da saurin kwararar ruwa sama da 33 µm/s. An lura da tarin maniyyi suna kusantowa da mannewa a gefen hanyoyin microfluidic don gujewa sharewa da saurin kwararar ruwa sama da 33 µm/s. Было zamечено, что пучки сперматозоидов приближаются избежать сметания со скоростью потока жидкости> 33 мкм / с. An lura da tarin maniyyi suna kusantowa da mannewa a gefen hanyoyin microfluidic don gujewa sharar ruwa a cikin ƙimar kwararar ruwa sama da 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 µ过.33 µm/s. Было zamечено, что пучки сперматозоидов приближаются избежать сметания потоком жидкости со скоростью > 33 мкм/с. An lura da tarin maniyyi suna kusantowa da mannewa a gefen tashar microfluidic don guje wa kwararar ruwa a sama da 33 µm/s.Binciken na'urar daukar hoto da kuma na'urar daukar hoto ta lantarki ta gano cewa tarin maniyyi sun sami tallafi daga kayan da ke da yawa. Bayanan da aka samu sun nuna motsi na musamman na spermatozoa na kaji Sharkazi, da kuma ikon spermatozoa na tara da kuma samar da fakitin motsi, wanda ke taimakawa wajen fahimtar adana maniyyi na dogon lokaci a cikin SMT.
Domin samun hadi a cikin mutane da yawancin dabbobi, maniyyi da ƙwai dole ne su isa wurin hadi a lokacin da ya dace. Saboda haka, dole ne a yi hadi kafin ko lokacin da za a yi ovulation. A gefe guda kuma, wasu dabbobi masu shayarwa, kamar karnuka, da kuma nau'ikan dabbobi marasa shayarwa, kamar kwari, kifi, dabbobi masu rarrafe, da tsuntsaye, suna adana maniyyi a cikin gabobin haihuwa na tsawon lokaci har sai ƙwainsu sun shirya don hadi (hadi asynchronous 1). Tsuntsaye suna iya kiyaye wanzuwar maniyyi wanda zai iya hadi ƙwai na tsawon makonni 2-102.
Wannan wata siffa ce ta musamman da ke bambanta tsuntsaye da sauran dabbobi, domin tana ba da damar yin hadi bayan an yi hadi sau ɗaya na tsawon makonni da dama ba tare da haɗuwa da ovulation a lokaci guda ba. Babban sashin ajiyar maniyyi, wanda ake kira tubule na ajiyar maniyyi (SST), yana cikin lanƙwasa na ciki a mahaɗin uterovaginal. Zuwa yanzu, ba a fahimci hanyoyin da maniyyi ke shiga, zama, da fita daga bankin maniyyi gaba ɗaya ba. Dangane da binciken da aka yi a baya, an gabatar da hasashe da yawa, amma babu ɗayansu da aka tabbatar.
Forman4 ya yi hasashen cewa maniyyi yana ci gaba da zama a cikin ramin SST ta hanyar ci gaba da motsi mai juyawa a kan alkiblar kwararar ruwa ta hanyoyin furotin da ke kan ƙwayoyin halittar SST (rheology). ATP yana raguwa saboda yawan aikin flagellar da ake buƙata don kiyaye maniyyi a cikin lumen na SST kuma motsi daga ƙarshe yana raguwa har sai an fitar da maniyyi daga bankin maniyyi ta hanyar kwararar ruwa kuma a fara sabuwar tafiya ta cikin bututun fallopian mai hawa don takin maniyyi. Kwai (Forman4). Wannan samfurin ajiyar maniyyi yana da goyan bayan gano ta hanyar immunocytochemistry na aquaporins 2, 3 da 9 da ke cikin ƙwayoyin halittar SST. Zuwa yanzu, bincike kan ilimin halittar maniyyi na kaza da rawar da yake takawa a cikin ajiyar SST, zaɓin maniyyi na farji, da gasar maniyyi ba su da yawa. A cikin kaji, maniyyi yana shiga farji bayan haɗuwa ta halitta, amma fiye da kashi 80% na maniyyi ana fitar da su daga farji jim kaɗan bayan haɗuwa. Wannan yana nuna cewa farji shine babban wurin zaɓin maniyyi a cikin tsuntsaye. Bugu da ƙari, an ruwaito cewa ƙasa da kashi 1% na maniyyi da aka haɗa a cikin farji yana ƙarewa a cikin SSTs2. A cikin yin allurar riga-kafi ga 'yan kaji a cikin farji, adadin maniyyi da ke kaiwa SST yakan ƙaru awanni 24 bayan yin allura. Zuwa yanzu, hanyar zaɓar maniyyi a lokacin wannan tsari ba a fayyace ba, kuma motsi na maniyyi na iya taka muhimmiyar rawa wajen ɗaukar maniyyi na SST. Saboda kauri da bangon bututun fallopian, yana da wuya a sa ido kai tsaye kan motsi na maniyyi a cikin bututun fallopian na tsuntsaye. Saboda haka, ba mu da ilimin asali game da yadda maniyyi ke canzawa zuwa SST bayan hadi.
An fahimci ilimin halittar jini a kwanan nan a matsayin muhimmin abu da ke sarrafa jigilar maniyyi a cikin al'aurar dabbobi masu shayarwa. Dangane da ikon maniyyi mai motsi don ƙaura a lokaci guda, Zaferani da abokan aikinsa8 sun yi amfani da tsarin corra microfluidic don ware maniyyi mai motsi daga samfuran maniyyi da aka rubuta. Wannan nau'in rarraba maniyyi yana da mahimmanci don maganin rashin haihuwa na likita da binciken asibiti, kuma an fi so fiye da hanyoyin gargajiya waɗanda ke ɗaukar lokaci da aiki kuma suna iya lalata yanayin maniyyi da ingancin tsarinsa. Duk da haka, har zuwa yau, babu wani bincike da aka gudanar kan tasirin fitar maniyyi daga al'aurar kaji akan motsi na maniyyi.
Ko da kuwa menene tsarin da ke kula da maniyyi da aka adana a cikin SST, masu bincike da yawa sun lura cewa spermatozoa masu zama suna haɗuwa kai da kai a cikin SST na kaji 9, 10, quails 2, da turkeys 11 don samar da tarin maniyyi masu haɗuwa. Marubutan sun ba da shawarar cewa akwai alaƙa tsakanin wannan haɗuwa da adana maniyyi na dogon lokaci a cikin SST.
Tingari da Lake12 sun ba da rahoton wata babbar alaƙa tsakanin maniyyi a cikin glandar da ke karɓar maniyyi na kaji, kuma sun yi tambaya ko maniyyi na tsuntsaye yana haɗuwa kamar yadda maniyyi na dabbobi masu shayarwa yake haɗuwa. Sun yi imanin cewa zurfafan haɗin da ke tsakanin maniyyi a cikin vas deferens na iya faruwa ne saboda damuwa da kasancewar adadi mai yawa na maniyyi a cikin ƙaramin sarari ke haifarwa.
Lokacin da ake kimanta halayen maniyyi a kan sabbin gilashin da aka rataye, ana iya ganin alamun haɗuwa na ɗan lokaci, musamman a gefunan ɗigon maniyyi. Duk da haka, haɗuwa sau da yawa yana damun aikin juyawa da ke da alaƙa da motsi mai ci gaba, wanda ke bayyana yanayin wannan lamari na ɗan lokaci. Masu binciken sun kuma lura cewa lokacin da aka ƙara mai narkewa a cikin maniyyi, ƙwayoyin halitta masu tsayi "kamar zare" sun bayyana.
An yi ƙoƙarin farko na kwaikwayon maniyyi ta hanyar cire siririn waya daga digon da aka rataye, wanda ya haifar da dogon vesicle mai kama da maniyyi da ya fito daga digon maniyyi. Nan da nan maniyyi ya yi layi a cikin vesicle, amma dukkan na'urar ta ɓace da sauri saboda ƙarancin 3D. Saboda haka, don nazarin haɗuwar maniyyi, ya zama dole a lura da motsi da halayen maniyyi kai tsaye a cikin bututun ajiyar maniyyi da aka keɓe, wanda yake da wahalar cimmawa. Saboda haka, ya zama dole a ƙirƙiri kayan aiki wanda ke kwaikwayon maniyyi don tallafawa nazarin motsi na maniyyi da halayyar haɗuwa. Brillard et al13 sun ba da rahoton cewa matsakaicin tsawon bututun ajiyar maniyyi a cikin 'yan kaji manya shine 400-600 µm, amma wasu SSTs na iya zama har zuwa 2000 µm. Mero da Ogasawara14 sun raba glandar seminiferous zuwa manyan tubules na ajiyar maniyyi da ba su da girma, waɗanda duka suna da tsayi iri ɗaya (~500 µm) da faɗin wuya (~38 µm), amma matsakaicin diamita na lumen na tubules shine 56.6 da 56.6 µm. , bi da bi 11.2 μm, bi da bi. A cikin binciken da ake yi yanzu, mun yi amfani da na'urar microfluidic mai girman tashoshi na 200 µm × 20 µm (W × H), wanda sashin giciyensa ya ɗan kusa da na SST da aka ƙara. Bugu da ƙari, mun bincika motsi na maniyyi da halayyar haɗakar maniyyi a cikin ruwa mai gudana, wanda ya yi daidai da hasashen Foreman cewa ruwa da ƙwayoyin epithelial na SST ke samarwa yana kiyaye maniyyi a cikin lumen a cikin alkiblar da ba ta da ƙarfi (rheological).
Manufar wannan binciken ita ce shawo kan matsalolin lura da motsin maniyyi a cikin bututun fallopian da kuma guje wa wahalhalun nazarin ilimin halittar maniyyi da halayyar maniyyi a cikin yanayi mai canzawa. An yi amfani da na'urar microfluidic wacce ke haifar da matsin lamba na hydrostatic don kwaikwayon motsi na maniyyi a cikin al'aurar kaza.
Idan aka ɗora digo na samfurin maniyyi mai narkewa (1:40) a cikin na'urar microchannel, ana iya gano nau'ikan motsi guda biyu na maniyyi (wani maniyyi da aka keɓe da maniyyi mai ɗaure). Bugu da ƙari, maniyyi yana yawan yin iyo akan halin yanzu (positive rheology; bidiyo na 1, 2). Duk da cewa tarin maniyyi yana da ƙarancin gudu fiye da na maniyyi mai kaɗaici (p < 0.001), sun ƙara yawan maniyyi da ke nuna rheotaxis mai kyau (p < 0.001; Tebur 2). Duk da cewa tarin maniyyi yana da ƙarancin gudu fiye da na maniyyi mai kaɗaici (p < 0.001), sun ƙara yawan maniyyi da ke nuna rheotaxis mai kyau (p < 0.001; Tebur 2). Хотя пучки сперматозоидов имели более процент сперматозоидов, демонстрирующих положительный реотаксис (p <0,001; таблица 2). Duk da cewa tarin maniyyi yana da ƙarancin gudu fiye da na maniyyi ɗaya (p < 0.001), sun ƙara yawan maniyyi wanda ke nuna rheotaxis mai kyau (p < 0.001; Tebur 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 速度 （p <0.001）百分比 （p <0.001 ： 2）） Хотя скорость пучков сперматозоидов была ниже. сперматозоидов с положительной реологией (p <0,001; таблица 2). Duk da cewa saurin tarin maniyyi ya yi ƙasa da na maniyyi ɗaya (p < 0.001), sun ƙara yawan maniyyi tare da rheology mai kyau (p < 0.001; Tebur 2).An kiyasta cewa rheology mai kyau ga maniyyi ɗaya da tufts yana da kusan kashi 53% da 85%, bi da bi.
An lura cewa maniyyin kaji sharkasi nan da nan bayan fitar maniyyi yana samar da tarin layi, wanda ya kunshi mutane da dama. Waɗannan tarin suna ƙaruwa da kauri akan lokaci kuma suna iya kasancewa a cikin vitro na tsawon sa'o'i da yawa kafin su watse (bidiyo na 3). Waɗannan tarin filamentous suna da siffar kamar echidna spermatozoa wanda ke samuwa a ƙarshen epididymis. An gano cewa maniyyin kaji Sharkashi yana da babban yanayin haɗuwa da samar da tarin reticulate cikin ƙasa da minti ɗaya bayan tattarawa. Waɗannan hasken suna da ƙarfi kuma suna iya mannewa akan duk wani bango ko abubuwa masu tsauri. Kodayake tarin maniyyi yana rage saurin ƙwayoyin maniyyi, a bayyane yake cewa ta hanyar macroscopic suna ƙara layinsu. Tsawon tarin ya bambanta dangane da adadin maniyyin da aka tattara a cikin tarin. An ware sassa biyu na tarin: ɓangaren farko, gami da kan maniyyin da aka haɗa, da ɓangaren ƙarshe, gami da wutsiya da ƙarshen maniyyin gaba ɗaya. Ta amfani da kyamarar sauri mai sauri (950 fps), an lura da kan maniyyi mai ɗauke da ƙwayoyin halitta a farkon ɓangaren, wanda ke da alhakin motsin ƙwayar saboda motsinsu na juyawa, yana jan sauran cikin ƙwayar tare da motsi mai kama da helical (Bidiyo na 4). Duk da haka, a cikin dogayen tufts, an lura cewa wasu kan maniyyi masu kyauta suna manne da jiki da kuma ɓangaren ƙarshe na tuft suna aiki azaman vanes don taimakawa wajen motsa tuft.
Yayin da suke cikin ruwa a hankali, tarin maniyyi suna tafiya a layi ɗaya da juna, duk da haka, suna fara haɗuwa da juna kuma suna manne da duk abin da yake a tsaye, don kada kwararar ruwa ta tafi da su yayin da saurin kwararar ke ƙaruwa. Ƙwayoyin suna samuwa lokacin da ƴan ƙwayoyin maniyyi suka kusanci juna, suna fara motsawa cikin daidaituwa kuma suna naɗe juna, sannan su manne da wani abu mai mannewa. Hotuna na 1 da 2 sun nuna yadda maniyyi ke kusantar juna, suna samar da haɗuwa yayin da wutsiyoyin ke naɗe juna.
Masu binciken sun yi amfani da matsin lamba na hydrostatic don samar da kwararar ruwa a cikin wani ƙaramin tashar don nazarin ilimin halittar maniyyi. An yi amfani da ƙaramin tashar mai girman 200 µm × 20 µm (W × H) kuma tsawon 3.6 µm. Yi amfani da ƙananan hanyoyin sadarwa tsakanin kwantena tare da sirinji da aka sanya a ƙarshen. An yi amfani da launin abinci don sa hanyoyin su bayyana sosai.
A ɗaure kebul da kayan haɗi zuwa bango. An ɗauki bidiyon da na'urar hangen nesa ta mataki. Tare da kowane hoto, an gabatar da na'urar hangen nesa ta mataki da hotunan taswirar. (A) Haɗin da ke tsakanin rafuka biyu yana hana kwararar ruwa saboda motsi mai haske (kibiya ja). (B) Haɗin da ke tsakanin bututun bututu da bangon tashar (kibiya ja), a lokaci guda an haɗa su da wasu ƙugiya biyu (kibiya rawaya). (C) Ƙwayoyin maniyyi a cikin tashar microfluidic suna fara haɗuwa da juna (kibiya ja), suna samar da raga na ƙugiya maniyyi. (D) Samar da hanyar sadarwa ta ƙugiya maniyyi.
Lokacin da aka ɗora digo na maniyyi mai narkewa a cikin na'urar microfluidic kuma aka ƙirƙiri kwarara, an lura da yadda maniyyin ke motsawa a kan alkiblar kwararar. Ƙwayoyin sun dace sosai da bangon ƙananan hanyoyin, kuma kawunan 'yantacce a ɓangaren farko na tarin sun dace sosai a kansu (bidiyo na 5). Suna kuma manne da duk wani barbashi da ke tsaye a hanyarsu, kamar tarkace, don hana kwararar ta tafi da su. Bayan lokaci, waɗannan tufts ɗin sun zama dogayen filaments waɗanda ke kama wasu spermatozoa guda ɗaya da ƙananan tufts (Bidiyo na 6). Yayin da kwararar ta fara raguwa, dogayen layukan maniyyi sun fara samar da hanyar sadarwa ta layukan maniyyi (Bidiyo na 7; Hoto na 2).
A saurin kwararar ruwa mai yawa (V > 33 µm/s), ana ƙara yawan motsin zare a matsayin yunƙurin kama tarin maniyyi daban-daban waɗanda suka fi ƙarfin kwararar ruwa. A saurin kwararar ruwa mai yawa (V > 33 µm/s), ana ƙara yawan motsin zare a matsayin yunƙurin kama tarin maniyyi daban-daban waɗanda suka fi ƙarfin kwararar ruwa. При высокой скорости потока (V > 33 мкм/с) спиралевидные множество отдельных сперматозоидов, образующих пучки, которые лучше противостоят дрейюющей A yawan kwararar ruwa mai yawa (V > 33 µm/s), motsin helical na zare yana ƙaruwa yayin da suke ƙoƙarin kama tarin maniyyi daban-daban waɗanda suka fi iya jure ƙarfin kwararar.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。在 高 流速 (v> 33 µm/s) 时 , 的 螺旋 展开地 抵抗 的 漂移力…………………………. При высоких скоростях потока (V > 33 мкм/с) спиральное движение сперматозоидов, образующих пучки, чтобы лучше сопротивляться силам дрейфа потока. A yawan kwararar ruwa mai yawa (V > 33 µm/s), motsin helical na zaruruwa yana ƙaruwa a ƙoƙarin kama tarin maniyyi da yawa don su fi tsayayya da ƙarfin kwararar ruwa.Sun kuma yi ƙoƙarin haɗa ƙananan tashoshi a bangon gefe.
An gano tarin maniyyi a matsayin tarin kan maniyyi da wutsiyoyin lanƙwasa ta amfani da na'urar hangen nesa mai haske (LM). An kuma gano tarin maniyyi masu tarin abubuwa daban-daban a matsayin kawunan da aka murɗe da tarin flagellar, wutsiyoyin maniyyi masu tarin yawa, kawunan maniyyi da aka haɗa da wutsiya, da kawunan maniyyi masu lanƙwasa a matsayin ƙwayoyin halitta masu tarin yawa. na'urar hangen nesa ta watsa electron (TEM). Binciken na'urar hangen nesa ta electron (SEM) ya nuna cewa tarin maniyyi tarin kawunan maniyyi ne kuma tarin maniyyi ya nuna hanyar sadarwa da aka haɗa ta wutsiya masu naɗewa.
An yi nazarin yanayin da kuma tsarin ƙwayayen maniyyi, samuwar ƙunƙunan maniyyi ta amfani da na'urar hangen nesa mai haske (rabin sashe), na'urar hangen nesa ta electron microscopy (SEM) da na'urar hangen nesa ta transmission electron microscopy (TEM), an yi wa maniyyi fenti da acridine orange sannan aka duba ta ta amfani da na'urar hangen nesa ta epifluorescence microscopy.
Tabon maniyyi da aka yi da acridine orange (Hoto na 3B) ya nuna cewa kan maniyyin ya manne tare kuma an rufe shi da kayan ɓoyewa, wanda ya haifar da samuwar manyan tufts (Hoto na 3D). Tabon maniyyin ya ƙunshi tarin maniyyi tare da hanyar sadarwa ta wutsiya da aka haɗa (Hoto na 4A-C). Tabon maniyyin ya ƙunshi wutsiyoyin maniyyi da yawa da aka manne tare (Hoto na 4D). Sirrin (Hoto na 4E,F) ya rufe kan tarin maniyyi.
Samar da tarin maniyyi Ta amfani da na'urar hangen nesa ta zamani da kuma na'urar hangen nesa ta acridine orange, an nuna cewa kawunan maniyyi sun manne tare. (A) Samar da maniyyi na farko yana farawa da maniyyi (da'irar fari) da maniyyi uku (da'irar rawaya), tare da karkacewa yana farawa daga wutsiya kuma yana ƙarewa a kai. (B) Hoton ma'aunin maniyyi mai launin acridine orange wanda ke nuna kan maniyyi mai manne (kibiyoyi). Fitar tana rufe kan(s). Girman × 1000. (C) Ci gaban babban katako da ake jigilar shi ta hanyar kwarara a cikin tashar microfluidic (ta amfani da kyamarar sauri mai sauri a 950 fps). (D) Micrograph na ma'aunin maniyyi mai launin acridine orange wanda ke nuna manyan tufts (kibiyoyi). Girman: ×200.
Na'urar daukar hoton lantarki ta hanyar amfani da hasken rana da kuma na'urar daukar hoton maniyyi da aka yi wa fenti da ruwan lemu mai siffar acridine. (A, B, D, E) na'urar daukar hoton launi ta hanyar amfani da hasken rana, kuma C da F na'urar daukar hoton maniyyi mai siffar acridine orange ce, wadda ke nuna hadewar maniyyi da yawa da ke nade gidan yanar gizo na caudal. (AC) Ana nuna tarin maniyyi a matsayin hanyar sadarwa ta wutsiyoyin da aka makala (kibiyoyi). (D) Mannewa da wasu maniyyi (tare da wani abu mai mannewa, ruwan hoda, kibiya) da ke nadewa a kusa da wutsiya. (E da F) Tarin kan maniyyi (masu nuna haske) da aka rufe da kayan mannewa (masu nuna haske). Tarin maniyyi ya samar da tarin maniyyi da siffofi masu kama da vortex da yawa (F). (C) ×400 da (F) ×200 girma.
Ta amfani da na'urar hangen nesa ta lantarki, mun gano cewa tarin maniyyi yana da wutsiya masu haɗe (Hoto na 6A, C), kawunan da aka haɗa da wutsiya (Hoto na 6B), ko kawunan da aka haɗa da wutsiya (Hoto na 6D). Kan maniyyi a cikin tarin suna lanƙwasa, suna bayyana a sashe na biyu na yankunan nukiliya (Hoto na 6D). A cikin tarin yanke, maniyyi yana da kai mai karkace tare da yankuna biyu na nukiliya da yankuna da yawa na flagellar (Hoto na 5A).
Na'urar daukar hoto ta lantarki mai launi ta dijital wadda ke nuna wutsiyoyin da ke haɗa maniyyi da kuma kayan da ke haɗa kan maniyyi. (A) Wutsiyar da aka haɗa da adadi mai yawa na maniyyi. Lura da yadda wutsiyar take a cikin hoton (kibiya) da kuma yanayin ƙasa (kibiya). (B) Kan (kibiya) na maniyyi an haɗa shi da wutsiya (kibiya). (C) An haɗa wutsiyoyin maniyyi da yawa (kibiya). (D) Kayan haɗa maniyyi (AS, shuɗi) sun haɗa kan maniyyi huɗu (shuɗi).
An yi amfani da na'urar daukar hoton lantarki ta hanyar amfani da na'urar daukar hoton lantarki don gano kan maniyyi a cikin tarin maniyyi da aka rufe da abubuwan da ke fita daga jiki ko membranes (Hoto na 6B), wanda ke nuna cewa tarin maniyyi an makale su da kayan da ke cikin jiki. An tattara kayan da aka haɗa a cikin kan maniyyi (jigon kai kamar na jellyfish; Hoto na 5B) kuma an faɗaɗa su nesa, suna ba da kamannin rawaya mai haske a ƙarƙashin na'urar daukar hoton haske lokacin da aka yi masa fenti da acridine orange (Hoto na 6C). Wannan abu yana bayyane a fili a ƙarƙashin na'urar daukar hoton haske kuma ana ɗaukarsa a matsayin abin ɗaurewa. Sassan rabin siriri (Hoto na 5C) da kuma smears ɗin maniyyi da aka yi wa fenti da acridine orange sun nuna tarin maniyyi da ke ɗauke da kan da aka cika da yawa da wutsiya masu lanƙwasa (Hoto na 5D).
Na'urorin daukar hoto daban-daban da ke nuna tarin kawunan maniyyi da wutsiyoyin da aka naɗe ta amfani da hanyoyi daban-daban. (A) Na'urar daukar hoto mai launi na dijital mai rarrabawa ta hanyar amfani da na'urar daukar hoto mai launi na wani tarin maniyyi da ke nuna kan maniyyi mai naɗewa tare da tsakiya mai sassa biyu (shuɗi) da sassan flagellar da dama (kore). (B) Na'urar daukar hoto mai launi ta dijital mai nuna tarin kawunan maniyyi masu kama da jellyfish (kibiyoyi) waɗanda suka bayyana a rufe. (C) Sashe mai siriri wanda ke nuna kawunan maniyyi masu tarin yawa (kibiyoyi) da wutsiyoyin da aka naɗe (kibiyoyi). (D) Na'urar daukar hoto mai launi ta maniyyi da aka yi wa fenti da acridine orange wanda ke nuna tarin kawunan maniyyi (kibiyoyi) da wutsiyoyin da aka naɗe (kibiyoyi). Lura cewa wani abu mai mannewa (S) ya rufe kan maniyyi. (D) × 1000 girma.
Ta amfani da na'urar hangen nesa ta hanyar watsawa (Hoto na 7A), an kuma lura cewa kawunan maniyyi sun karkace kuma ƙwayoyin halittar suna da siffar karkace, kamar yadda aka tabbatar ta hanyar smears ɗin maniyyi da aka yi wa fenti da acridine orange kuma aka duba ta amfani da na'urar hangen nesa ta fluorescence (Hoto na 7B).
(A) Ma'aunin watsa launi na dijital na lantarki da (B) Ma'aunin ruwan lemu na Acridine wanda ke nuna kawunan maniyyi da haɗewar kawunan maniyyi da wutsiyoyinsa (kibiyoyi). (B) × 1000 girma.
Wani abin sha'awa da aka gano shi ne cewa tarin maniyyin Sharkazi yana samar da tarin filamentous masu motsi. Halayen waɗannan tarin suna ba mu damar fahimtar rawar da za su iya takawa wajen sha da adana maniyyi a cikin SST.
Bayan haɗuwa, maniyyi yana shiga farji kuma yana yin tsari mai zurfi na zaɓi, wanda ke haifar da ƙarancin adadin maniyyi da ke shiga SST15,16. Zuwa yanzu, hanyoyin da maniyyi ke shiga da fita SST ba su da tabbas. A cikin kaji, ana adana maniyyi a cikin SST na tsawon makonni 2 zuwa 10, ya danganta da nau'in 6. Ana ci gaba da cece-kuce game da yanayin maniyyi yayin ajiya a cikin SST. Shin suna cikin motsi ko suna hutawa? A wata ma'anar, ta yaya ƙwayoyin maniyyi ke riƙe matsayinsu a cikin SST na dogon lokaci?
Forman4 ya ba da shawarar cewa za a iya bayanin wurin zama na SST da kuma fitar da maniyyi ta hanyar amfani da motsin maniyyi. Marubutan sun yi hasashen cewa maniyyi yana riƙe matsayinsa ta hanyar yin iyo a kan kwararar ruwa da SST epithelium ya samar kuma maniyyi yana fitar da maniyyi daga SST lokacin da saurinsa ya faɗi ƙasa da inda suka fara komawa baya saboda rashin kuzari. Zaniboni5 ya tabbatar da kasancewar aquaporins 2, 3, da 9 a cikin ɓangaren apical na ƙwayoyin epithelial na SST, wanda zai iya tallafawa samfurin ajiyar maniyyi na Foreman a kaikaice. A cikin binciken da aka yi a yanzu, mun gano cewa kusan rabin maniyyin Sharkashi yana nuna ingantaccen rheology a cikin ruwan da ke gudana, kuma cewa tarin maniyyi da aka haɗa yana ƙara yawan maniyyi da ke nuna ingantaccen rheology, kodayake haɗuwa yana rage su. Yadda ƙwayoyin maniyyi ke tafiya ta cikin bututun fallopian na tsuntsu zuwa wurin da aka hadi ba a fahimci shi sosai ba. A cikin dabbobi masu shayarwa, ruwan follicular yana jan hankalin maniyyi. Duk da haka, ana kyautata zaton cewa masu jan hankalin maniyyi suna jagorantar maniyyi ya kusanci nisan nesa7. Saboda haka, wasu hanyoyin suna da alhakin jigilar maniyyi. An ruwaito cewa ikon maniyyi na karkatar da ruwa da gudana a kan ruwan bututun fallopian da aka saki bayan haɗuwa shine babban abin da ke haifar da nisantar maniyyi a cikin beraye. Parker 17 ya ba da shawarar cewa maniyyi ya ratsa ƙwayoyin halittar ta hanyar yin iyo a kan kwararar ciliary a cikin tsuntsaye da dabbobi masu rarrafe. Kodayake ba a nuna shi ta hanyar gwaji a cikin tsuntsaye ba, Adolphi18 shine farkon wanda ya gano cewa maniyyin tsuntsaye yana ba da sakamako mai kyau lokacin da aka ƙirƙiri siririn Layer na ruwa tsakanin murfin murfin da zamewa tare da zamewa da zamewa. Rheology. Hino da Yanagimachi [19] sun sanya mahaɗin ƙwai-tubal-uterine na linzamin kwamfuta a cikin zoben perfusion kuma suka allurar µl na tawada a cikin isthmus don ganin yadda ruwa ke gudana a cikin bututun fallopian. Sun lura da motsi mai aiki na matsewa da shakatawa a cikin bututun fallopian, wanda duk ƙwallon tawada suna motsawa akai-akai zuwa ampulla na bututun fallopian. Marubutan sun jaddada muhimmancin kwararar ruwan bututu daga ƙananan bututun fallopian zuwa sama don ɗaga maniyyi da hadi. Brillard20 ta ruwaito cewa a cikin kaji da turkeys, spermatozoa suna ƙaura ta hanyar motsi mai aiki daga ƙofar farji, inda ake adana su, zuwa mahaɗin mahaifa da farji, inda ake adana su. Duk da haka, wannan motsi ba a buƙatar tsakanin mahaɗin uterovaginal da infundibulum saboda ana jigilar spermatozoa ta hanyar ƙaura mara aiki. Sanin waɗannan shawarwari na baya da sakamakon da aka samu a cikin binciken da ake yi yanzu, za a iya ɗauka cewa ikon spermatozoa na motsawa sama (rheology) yana ɗaya daga cikin halayen da tsarin zaɓi ya dogara da su. Wannan yana ƙayyade wucewar spermatozoa ta farji da shigar su cikin CCT don ajiya. Kamar yadda Forman4 ya ba da shawara, wannan na iya sauƙaƙe tsarin shigar da maniyyi SST da mazauninsa na ɗan lokaci sannan ya fita lokacin da saurinsu ya fara raguwa.
A gefe guda kuma, Matsuzaki da Sasanami 21 sun ba da shawarar cewa maniyyin tsuntsaye yana fuskantar canje-canje a motsi daga barci zuwa motsi a cikin hanyoyin haihuwa na maza da mata. An gabatar da hana motsi na maniyyi a cikin SST don bayyana tsawon lokacin ajiya na maniyyi sannan kuma sake farfaɗowa bayan barin SST. A ƙarƙashin yanayin hypoxic, Matsuzaki da abokan aikinsa 1 sun ba da rahoton yawan samarwa da sakin lactate a cikin SST, wanda zai iya haifar da hana motsi na maniyyi. A wannan yanayin, mahimmancin ilimin halittar maniyyi yana bayyana a cikin zaɓar da shan maniyyi, ba a cikin ajiyar su ba.
Tsarin tattara maniyyi ana ɗaukarsa a matsayin wani abu mai yiwuwa ga tsawon lokacin adana maniyyi a cikin SST, domin wannan tsari ne da aka saba gani na riƙe maniyyi a cikin kaji2,22,23. Bakst da abokan aikinsa 2 sun lura cewa yawancin maniyyi suna manne da juna, suna samar da tarin maniyyi, kuma ba kasafai ake samun maniyyi guda ɗaya a cikin CCM na kwarkwata ba. A gefe guda kuma, Wen da abokan aikinsa 24 sun lura da ƙarin maniyyi da aka warwatse da ƙarancin tarin maniyyi a cikin lumen na SST a cikin kaji. Dangane da waɗannan abubuwan da aka lura, ana iya ɗauka cewa sha'awar tattara maniyyi ya bambanta tsakanin tsuntsaye da tsakanin maniyyi a cikin wannan fitar maniyyi. Bugu da ƙari, Van Krey da abokan aikinsa 9 sun ba da shawarar cewa rabuwar maniyyi da aka haɗa bazuwar shine ke haifar da shigar maniyyi a hankali cikin lumen na bututun fallopian. A cewar wannan hasashe, ya kamata a fara fitar da maniyyi da ƙarancin ƙarfin tattara maniyyi daga SST. A wannan mahallin, ikon maniyyi na tara ruwa na iya zama wani abu da ke tasiri ga sakamakon gasar maniyyi a cikin tsuntsaye masu datti. Bugu da ƙari, tsawon lokacin da maniyyin da aka tara ruwa ya rabu, tsawon lokacin da aka samu haihuwa zai ci gaba.
Duk da cewa an lura da tarin maniyyi da taruwarsa cikin tarin a cikin bincike da dama2,22,24, ba a bayyana su dalla-dalla ba saboda sarkakiyar lura da kinematic ɗinsu a cikin SST. An yi ƙoƙari da yawa don nazarin taruwar maniyyi a cikin vitro. An lura da taruwar maniyyi mai yawa amma mai ɗan lokaci lokacin da aka cire siririn waya daga digon iri da ke rataye. Wannan ya haifar da gaskiyar cewa wani kumfa mai tsayi ya fito daga digon, yana kwaikwayon glandar maniyyi. Saboda iyakokin 3D da gajeren lokacin bushewar digo, dukkan tubalin ya faɗi cikin matsala9. A cikin binciken da ake yi a yanzu, ta amfani da kaji Sharkashi da ƙananan kwakwalwan microfluidic, mun sami damar bayyana yadda waɗannan tufts ke samuwa da kuma yadda suke motsawa. Tarin maniyyi sun samo asali nan da nan bayan tattara maniyyi kuma an gano suna motsawa a cikin karkace, suna nuna rheology mai kyau lokacin da suke cikin kwararar. Bugu da ƙari, lokacin da aka duba ta hanyar macroscopic, an lura da tarin maniyyi don ƙara layin motsi idan aka kwatanta da spermatozoa da aka ware. Wannan yana nuna cewa haɗuwar maniyyi na iya faruwa kafin shigar SST kuma samar da maniyyi ba ta takaita ga ƙaramin yanki ba saboda damuwa kamar yadda aka nuna a baya (Tingari da Lake12). A lokacin samuwar tuft, maniyyi yana iyo a lokaci guda har sai sun samar da mahaɗi, sannan wutsiyoyinsu suna naɗe juna kuma kan maniyyi yana nan ba tare da wata matsala ba, amma wutsiya da ɓangaren nesa na maniyyi suna manne tare da wani abu mai mannewa. Saboda haka, kan jijiyar da ba ta da matsala yana da alhakin motsi, yana jan sauran jijiyar. Binciken na'urar daukar hoto ta electron microscopy na tarin maniyyi ya nuna kawunan maniyyi da aka haɗa da aka rufe da kayan manne da yawa, yana nuna cewa kawunan maniyyi an haɗa su a cikin maƙullan hutawa, wanda wataƙila ya faru bayan isa wurin ajiya (SST).
Idan aka yi wa maniyyi fenti da acridine orange, ana iya ganin kayan manne na waje da ke kewaye da ƙwayoyin maniyyi a ƙarƙashin na'urar hangen nesa mai haske. Wannan abu yana ba da damar mannewar maniyyi ya manne da kuma manne wa duk wani saman ko barbashi da ke kewaye don kada su zame tare da kwararar da ke kewaye. Don haka, abubuwan da muka lura sun nuna rawar da mannewar maniyyi ke takawa a cikin nau'in manne mai motsi. Ikonsu na yin iyo a kan kwararar ruwa da kuma mannewa a saman da ke kusa yana ba da damar maniyyi ya daɗe a cikin SST.
Rothschild25 ta yi amfani da kyamarar hemocytometry don nazarin rarraba maniyyi na shanu a cikin digo na dakatarwa, tana ɗaukar hotunan micrographs ta cikin kyamara tare da madaidaiciyar axis na gani na madubin. Sakamakon ya nuna cewa maniyyi ya jawo hankalin saman ɗakin. Marubutan sun ba da shawarar cewa akwai yiwuwar hulɗar hydrodynamic tsakanin maniyyi da saman. Idan aka yi la'akari da wannan, tare da ikon maniyyin kazar Sharkashi na samar da tufts masu mannewa, yana iya ƙara yiwuwar maniyyi ya manne da bangon SST kuma a adana shi na dogon lokaci.
Bccetti da Afzeliu26 sun ruwaito cewa ana buƙatar glycocalyx na maniyyi don gane kwayoyin halitta da kuma haɗa su. Forman10 sun lura cewa hydrolysis na α-glycosidic bonds a cikin glycoprotein-glycolipid coatings ta hanyar magance maniyyi na tsuntsaye da neuraminidase ya haifar da raguwar haihuwa ba tare da shafar motsi na maniyyi ba. Marubutan sun ba da shawarar cewa tasirin neuraminidase akan glycocalyx yana lalata sequestration na maniyyi a mahaɗin mahaifa da farji, ta haka ne rage haihuwa. Abubuwan da suka lura ba za su iya yin watsi da yuwuwar cewa maganin neuraminidase na iya rage gane maniyyi da kuma ugcyte ba. Forman da Engel10 sun gano cewa haihuwa ta ragu lokacin da aka yi wa kaji haihuwa ta hanyar amfani da maniyyi da aka yi wa magani da neuraminidase. Duk da haka, IVF tare da maniyyi da aka yi wa magani da neuraminidase bai shafi haihuwa ba idan aka kwatanta da kaji masu sarrafawa. Marubutan sun kammala da cewa canje-canje a cikin rufin glycoprotein-glycolipid da ke kewaye da membrane na maniyyi ya rage ikon maniyyi na yin taki ta hanyar rage jerin maniyyi a wurin haɗuwar mahaifa da farji, wanda hakan ya ƙara asarar maniyyi saboda saurin haɗuwar mahaifa da farji, amma ba ya shafar gano maniyyi da kuma gane ƙwai.
A cikin turkeys, Bakst da Bauchan 11 sun sami ƙananan vesicles da gutsuttsuran membrane a cikin lumen na SST kuma sun lura cewa wasu daga cikin waɗannan granules sun haɗu da membrane na maniyyi. Marubutan sun ba da shawarar cewa waɗannan alaƙar na iya taimakawa wajen adana maniyyi na dogon lokaci a cikin SST. Duk da haka, masu binciken ba su ƙayyade tushen waɗannan barbashi ba, ko ƙwayoyin epithelial na CCT ne suka fitar da su, ko tsarin haihuwa na maza ya samar kuma ya fitar da su, ko kuma maniyyin da kansa ya samar. Hakanan, waɗannan barbashi suna da alhakin haɗuwa. Grützner et al27 sun ba da rahoton cewa ƙwayoyin epithelial na epididymal suna samarwa da kuma fitar da wani takamaiman furotin wanda ake buƙata don ƙirƙirar sassan seminal guda ɗaya. Marubutan sun kuma ba da rahoton cewa watsar waɗannan fakitin ya dogara ne akan hulɗar sunadarai na epididymal. Nixon et al28 sun gano cewa adnexa yana fitar da furotin, osteonectin mai acidic mai arzikin cysteine; SPARC yana da hannu wajen ƙirƙirar tufts na maniyyi a cikin echidnas da platypuses masu gajeru. Yaɗuwar waɗannan hasken yana da alaƙa da asarar wannan furotin.
A cikin wannan binciken, binciken ultrastructural ta amfani da na'urar hangen nesa ta electron ya nuna cewa spermatozoa ya manne da wani abu mai yawa. Ana tsammanin waɗannan abubuwan ne ke haifar da haɗuwar da ke taruwa tsakanin da kewayen kawunan da ke manne, amma a ƙananan yawansu a yankin wutsiya. Muna ɗauka cewa wannan abu mai haɗuwa yana fitowa daga tsarin haihuwa na namiji (epididymis ko vas deferens) tare da maniyyi, tunda sau da yawa muna ganin maniyyi yana rabuwa daga lymph da jini yayin fitar maniyyi. An ruwaito cewa yayin da spermatozoa na tsuntsaye ke ratsawa ta cikin epididymis da vas deferens, suna fuskantar canje-canje masu alaƙa da balaga waɗanda ke tallafawa ikonsu na ɗaure sunadaran da kuma samun glycoproteins masu alaƙa da plasma lemma. Dorewar waɗannan sunadaran akan membranes na maniyyi a cikin SST yana nuna cewa waɗannan sunadaran na iya yin tasiri ga samun kwanciyar hankali na membrane na maniyyi 30 da kuma tantance haihuwa 31. Ahmad et al32 sun ruwaito cewa maniyyi da aka samu daga sassa daban-daban na tsarin haihuwa na maza (daga ƙwai zuwa ga ƙwai na distal) ya nuna ƙaruwar rayuwa a ƙarƙashin yanayin ajiyar ruwa, ba tare da la'akari da yanayin ajiya ba, kuma rayuwa a cikin kaji yana ƙaruwa a cikin bututun fallopian bayan an yi wa jarirai kwai.
Tufts na maniyyi na kaji Sharkashi suna da halaye da ayyuka daban-daban fiye da sauran nau'ikan halittu kamar echidnas, platypuses, beraye na itace, berayen barewa, da aladu na Guinea. A cikin kaji sharkasi, samuwar tarin maniyyi ya rage saurin iyo idan aka kwatanta da spermatozoa guda ɗaya. Duk da haka, waɗannan tarin sun ƙara yawan spermatozoa masu kyau a rheologically kuma sun ƙara ikon spermatozoa don daidaita kansu a cikin yanayi mai ƙarfi. Don haka, sakamakonmu ya tabbatar da shawarar da ta gabata cewa haɗuwar maniyyi a cikin SST yana da alaƙa da ajiyar maniyyi na dogon lokaci. Hakanan muna hasashen cewa saurin maniyyi don samar da tufts na iya sarrafa ƙimar asarar maniyyi a cikin SST, wanda zai iya canza sakamakon gasar maniyyi. A cewar wannan zato, spermatozoa tare da ƙarancin ƙarfin haɗuwa yana fitar da SST da farko, yayin da spermatozoa tare da babban ƙarfin haɗuwa yana samar da yawancin zuriya. Samuwar tarin maniyyi mai ramuka ɗaya yana da amfani kuma yana shafar rabon iyaye da yara, amma yana amfani da wata hanya daban. A cikin echidnas da platypuses, an shirya spermatozoa a layi ɗaya don ƙara saurin gaba na hasken. Kunshin echidnas suna motsawa sau uku fiye da maniyyi ɗaya. Ana kyautata zaton cewa samuwar irin waɗannan tufts na maniyyi a cikin echidnas daidaitawa ce ta juyin halitta don ci gaba da mamayewa, tunda mata ba sa yin jima'i kuma galibi suna haɗuwa da maza da yawa. Saboda haka, spermatozoa daga maniyyi daban-daban suna fafatawa sosai don hadi na ƙwai.
Ana iya ganin maniyyi mai girman gaske na kaji sharksi ta hanyar amfani da na'urar hangen nesa ta zamani (phase contrast microscopy), wanda ake ganin yana da amfani domin yana ba da damar yin nazari mai sauƙi kan halayen maniyyi a cikin vitro. Hanyar da samuwar maniyyi ke haifar da haihuwa a cikin kaji sharksi shi ma ya bambanta da wanda ake gani a wasu dabbobi masu shayarwa na mahaifa waɗanda ke wakiltar halayen maniyyi na haɗin gwiwa kamar beraye na itace, inda wasu maniyyi ke kaiwa ga ƙwai, yana taimaka wa wasu mutane masu alaƙa su isa da lalata ƙwai. don tabbatar da kansu. ɗabi'ar son kai. Haihuwa kai 34. Wani misali na halayyar haɗin gwiwa a cikin maniyyi an samo shi a cikin beraye na barewa, inda maniyyi ya sami damar gano da haɗuwa da maniyyi mafi alaƙa da kwayoyin halitta kuma ya samar da ƙungiyoyin haɗin gwiwa don ƙara saurin su idan aka kwatanta da maniyyi mara alaƙa35.
Sakamakon da aka samu a cikin wannan binciken bai saɓa wa ka'idar Foman ta adana maniyyi na dogon lokaci a cikin SWS ba. Masu binciken sun ba da rahoton cewa ƙwayoyin maniyyi suna ci gaba da motsawa a cikin kwararar ƙwayoyin epithelial da ke rufe SST na tsawon lokaci, kuma bayan wani lokaci, ajiyar makamashin ƙwayoyin maniyyi ya ƙare, wanda ke haifar da raguwar gudu, wanda ke ba da damar fitar da ƙananan abubuwa masu nauyin kwayoyin halitta. kuzarin maniyyi tare da kwararar ruwa daga lumen na SST. Kogon bututun fallopian. A cikin binciken da ake yi yanzu, mun lura cewa rabin maniyyi ɗaya ya nuna ikon yin iyo a kan ruwa mai gudana, kuma mannewarsu a cikin tarin ya ƙara ƙarfinsu na nuna rheology mai kyau. Bugu da ƙari, bayananmu sun yi daidai da na Matsuzaki et al. 1 waɗanda suka ba da rahoton cewa ƙaruwar fitar lactate a cikin SST na iya hana motsi na maniyyi. Duk da haka, sakamakonmu ya bayyana samuwar ligaments na motsi na maniyyi da halayensu na rheological a gaban yanayi mai ƙarfi a cikin microchannel a ƙoƙarin bayyana halayensu a cikin SST. Binciken da za a yi nan gaba zai iya mayar da hankali kan tantance sinadaran da asalin sinadarin da ke haifar da maye, wanda babu shakka zai taimaka wa masu bincike wajen ƙirƙiro sabbin hanyoyin adana ruwayen maniyyi da kuma ƙara tsawon lokacin haihuwa.
An zaɓi tsuntsayen sharkasi maza masu tsawon makonni 30 masu wuyan da ba su da wuya (homozygous dominant; Na Na) a matsayin masu bayar da maniyyi a cikin binciken. An kiwon tsuntsayen a Gonar Kaji ta Bincike ta Faculty of Agriculture, Jami'ar Ashit, Ashit Governorate, Masar. An ajiye tsuntsayen a cikin keji daban-daban (30 x 40 x 40 cm), an ba su shirin haske (awanni 16 na haske da awanni 8 na duhu) kuma an ciyar da su da abinci mai ɗauke da gram 160 na furotin mai ɗanɗano, 2800 kcal na kuzarin da za a iya narkewa, gram 35 na calcium kowanne. gram 5 na phosphorus da ake samu a kowace kilogiram na abinci.
A cewar bayanai na 36, ​​​​37, an tattara maniyyi daga maza ta hanyar tausa ciki. An tattara jimillar samfuran maniyyi 45 daga maza 15 a cikin kwanaki 3. An narkar da maniyyi (n = 15/rana) nan da nan tare da ruwan diluent na Belsville Poultry Semen Diluent, wanda ya ƙunshi 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) da H2O (100 ml), pH = 7, 5, osmolarity 333 mOsm/kg38. An fara duba samfuran maniyyi da aka narkar da su a ƙarƙashin na'urar hangen nesa mai sauƙi don tabbatar da ingancin maniyyi (danshi) sannan a adana su a cikin ruwan wanka a zafin digiri 37 na Celsius har sai an yi amfani da su cikin rabin sa'a bayan an tattara su.
An bayyana tsarin kinematics da rheology na spermatozoa ta amfani da tsarin na'urorin microfluidic. An ƙara narkar da samfuran maniyyi zuwa 1:40 a cikin Beltsville Avian Avian Maniyyi Diluent, an ɗora shi a cikin na'urar microfluidic (duba ƙasa), kuma an ƙayyade sigogin motsi ta amfani da tsarin Computerized Maniyyi Analysis (CASA) da aka ƙirƙira a baya don halayyar microfluidics. akan motsi na spermatozoa a cikin kafofin ruwa (Sashen Injiniyan Injiniya, Faculty of Engineering, Jami'ar Assiut, Masar). Ana iya sauke plugin ɗin a: http://www.assiutmicrofluidics.com/research/casa39. An auna saurin lanƙwasa (VCL, μm/s), saurin layi (VSL, μm/s) da matsakaicin saurin hanya (VAP, μm/s). An ɗauki bidiyon spermatozoa ta amfani da na'urar hangen nesa ta Optika XDS-3 mai juyawa (tare da manufa 40x) da aka haɗa da kyamarar Tucson ISH1000 a 30 fps na tsawon daƙiƙa 3. Yi amfani da manhajar CASA don yin nazarin aƙalla wurare uku da hanyoyin maniyyi 500 a kowane samfuri. An sarrafa bidiyon da aka ɗauka ta amfani da CASA na gida. Ma'anar motsi a cikin plug-in na CASA ya dogara ne akan saurin ninkaya na maniyyi idan aka kwatanta da yawan kwararar, kuma bai haɗa da wasu sigogi kamar motsi gefe-da-gefe ba, domin an gano cewa wannan ya fi aminci a cikin kwararar ruwa. An bayyana motsin rheological a matsayin motsi na ƙwayoyin maniyyi a kan alkiblar kwararar ruwa. An raba maniyyi tare da halayen rheological da adadin maniyyi mai motsi; an cire maniyyi waɗanda ke hutawa kuma suna motsi a cikin convective daga ƙidaya.
An samo dukkan sinadarai da aka yi amfani da su daga Elgomhoria Pharmaceuticals (Cairo, Egypt) sai dai idan an lura da wani abu daban. An ƙera na'urar kamar yadda El-sherry et al. 40 suka bayyana tare da wasu gyare-gyare. Kayan da aka yi amfani da su don ƙera ƙananan tashoshi sun haɗa da faranti na gilashi (Howard Glass, Worcester, MA), SU-8-25 negative resist (MicroChem, Newton, CA), diacetone alcohol (Sigma Aldrich, Steinheim, Germany), da polyacetone. -184, Dow Corning, Midland, Michigan). Ana ƙera ƙananan tashoshi ta amfani da lithography mai laushi. Da farko, an buga abin rufe fuska mai kariya mai haske tare da ƙirar microchannel da ake so akan firinta mai ƙuduri mai girma (Prismatic, Cairo, Egypt and Pacific Arts and Design, Markham, ON). An yi manyan samfuran ta amfani da faranti na gilashi a matsayin substrates. An tsaftace faranti a cikin acetone, isopropanol da ruwan deionized sannan aka shafa su da Layer na 20 µm na SU8-25 ta hanyar juyi (3000 rpm, minti 1). Bayan an busar da layukan SU-8 a hankali (65°C, minti 2 da 95°C, minti 10) sannan a fallasa su ga hasken UV na daƙiƙa 50. Bayan an fallasa su, a gasa su a zafin 65°C da 95°C na tsawon minti 1 da minti 4 don haɗa layukan SU-8 da aka fallasa, sannan a ci gaba da ƙara musu barasar diacetone na tsawon minti 6.5. A gasa waffles ɗin sosai (200°C na tsawon minti 15) don ƙara ƙarfafa layin SU-8.
An shirya PDMS ta hanyar haɗa monomer da hardener a cikin rabon nauyi na 10:1, sannan a cire gas ɗin a cikin injin cire iskar gas sannan a zuba a kan babban firam ɗin SU-8. An wartsake PDMS a cikin tanda (120°C, minti 30), sannan aka yanke hanyoyin, aka raba su da master, sannan aka huda su don ba da damar a haɗa bututu a wurin shiga da fita na microchannel. A ƙarshe, an haɗa ƙananan tashoshi na PDMS har abada zuwa faifai na microscope ta amfani da na'urar sarrafawa ta corona mai ɗaukuwa (Electro-Technic Products, Chicago, IL) kamar yadda aka bayyana a wani wuri. Microchannel ɗin da aka yi amfani da shi a wannan binciken yana da girman 200 µm × 20 µm (W × H) kuma yana da tsawon cm 3.6.
Ana samun kwararar ruwa da matsin lamba na hydrostatic ke haifarwa a cikin tashar microchannel ta hanyar kiyaye matakin ruwa a cikin ma'ajiyar shiga sama da bambancin tsayi Δh39 a cikin ma'ajiyar fitarwa (Hoto na 1).
inda f shine ma'aunin gogayya, wanda aka ayyana a matsayin f = C/Re don kwararar laminar a cikin tashar murabba'i mai kusurwa huɗu, inda C yake da daidaito dangane da rabon yanayin tashar, L shine tsawon tashar micro, Vav shine matsakaicin saurin da ke cikin tashar micro, Dh shine diamita na hydraulic na tashar, g - hanzarta nauyi. Ta amfani da wannan lissafi, ana iya ƙididdige matsakaicin saurin tashar ta amfani da lissafi mai zuwa: