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Haihuwar tsuntsaye ya dogara ne akan iyawarsu ta adana isasshiyar maniyyi mai ƙarfi na tsawon lokaci a cikin tubules ɗin ajiyar maniyyi (SST).Haƙiƙanin hanyar da maniyyi ya shiga, zama a ciki, da barin SST ya kasance mai kawo rigima.Maniyyi na sharkasi hens ya nuna babban hali zuwa agglutination, samar da wayar filamentous daure dauke da yawa cell.Saboda wahalar lura da motsi da halayyar maniyyi a cikin bututun fallopian mara kyau, mun yi amfani da na'urar microfluidic tare da sashin giciye na microchannel mai kama da na spermatozoa don nazarin spermatozoa agglutination da motsi.Wannan binciken ya tattauna yadda ɗimbin maniyyi ke samuwa, yadda suke motsawa, da yuwuwar rawar da za su iya takawa wajen faɗaɗa zaman maniyyi a cikin SST.Mun bincika saurin maniyyi da halayen rheological lokacin da aka haifar da kwararar ruwa a cikin tashar microfluidic ta matsin lamba na hydrostatic (yawan kwarara = 33 µm/s).Maniyyi yakan yi iyo a kan halin yanzu (tabbataccen rheology) kuma saurin daurin spermatozoon ya ragu sosai idan aka kwatanta da guda spermatozoa.An lura da tarin maniyyi suna motsawa cikin karkace kuma suna ƙaruwa da tsayi da kauri yayin da ake ɗaukar ƙarin maniyyi ɗaya. An lura da tarin maniyyi suna gabatowa kuma suna manne da bangon bangon tashoshi na microfluidic don gujewa sharewa da saurin kwararar ruwa> 33 µm/s. An lura da tarin maniyyi suna gabatowa kuma suna manne da bangon bangon tashoshi na microfluidic don gujewa sharewa da saurin kwararar ruwa> 33 µm/s. Было zamечено, что пучки сперматозоидов приближаются. ть сметания со скоростью потока жидкости> 33 мкм / с. An lura da tarin maniyyi don kusanci da mannewa ga bangon gefen tashoshi na microfluidic don gujewa sharewa a yawan kwararar ruwa> 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 ь сметания потоком жидкости со скоростью > 33 мкм/с. An lura da tarin maniyyi don kusanci da kuma manne da bangon gefen tashar microfluidic don guje wa zubar da ruwa a> 33 µm/s.Dubawa da watsawa da ƙananan ƙwayoyin lantarki sun nuna cewa tarin maniyyi suna da goyan bayan abubuwa masu yawa.Bayanan da aka samu sun nuna nau'in motsi na musamman na Sharkazi kaza spermatozoa, da kuma ikon spermatozoa don agglutinate da samar da nau'o'in wayar hannu, wanda ke taimakawa wajen fahimtar dogon lokaci na ajiyar maniyyi a cikin SMT.
Don cimma hadi a cikin mutane da yawancin dabbobi, dole ne maniyyi da ƙwai su isa wurin da ake hadi a lokacin da ya dace.Saboda haka, mating dole ne ya faru kafin ko a lokacin ovulation.A daya bangaren kuma, wasu dabbobi masu shayarwa, irin su karnuka, da nau’in da ba na dabbobi ba, kamar kwari, kifi, dabbobi masu rarrafe, da tsuntsaye, suna adana maniyyi a cikin gabobin haihuwarsu na tsawon lokaci har sai kwan su ya shirya don hadi (asynchronous fertilization 1).Tsuntsaye suna iya kula da yuwuwar spermatozoa masu iya takin ƙwai na makonni 2-102.
Wannan siffa ce ta musamman wacce ke bambanta tsuntsaye da sauran dabbobi, saboda yana ba da babbar yuwuwar hadi bayan fure ɗaya na makonni da yawa ba tare da saduwa da juna ba tare da kwai.Babban sashin ajiyar maniyyi, wanda ake kira tubule sperm storage tubule (SST), yana cikin folds na mucosal na ciki a mahadar uterovaginal.Har ya zuwa yau, ba a fahimci hanyoyin da maniyyi ke shiga, zama, da fita daga bankin maniyyi ba.Bisa binciken da aka yi a baya, an gabatar da zato da yawa, amma ba a tabbatar da ko daya daga cikinsu ba.
Forman4 yayi tsammanin cewa maniyyi yana kula da mazaunin su a cikin kogon SST ta hanyar ci gaba da motsi na oscillatory a kan jagorancin ruwa mai gudana ta hanyar tashoshin furotin da ke kan SST epithelial cell (rheology).ATP yana raguwa saboda yawan aiki na tuta da ake buƙata don kiyaye maniyyi a cikin SST lumen kuma motsin motsi daga ƙarshe ya ragu har sai an fitar da maniyyi daga bankin maniyyi ta hanyar ruwa kuma ya fara sabon tafiya zuwa hawan hawan fallopian don takin maniyyi.Kwai (Forman4).Wannan samfurin ajiyar maniyyi yana goyan bayan ganowa ta hanyar immunocytochemistry na aquaporins 2, 3 da 9 da ke cikin SST epithelial sel.Har ya zuwa yau, binciken da aka yi kan rheology na maniyyi kaji da rawar da yake takawa a cikin ajiyar SST, zaɓin maniyyi na farji, da gasar maniyyi ba a samu ba.A cikin kaji, maniyyi yana shiga cikin farji bayan dabi'ar dabi'a, amma fiye da 80% na spermatozoa ana fitar da su daga farji jim kadan bayan saduwa.Wannan yana nuna cewa farji shine wuri na farko don zaɓin maniyyi a cikin tsuntsaye.Bugu da kari, an bayar da rahoton cewa kasa da 1% na spermatozoa da aka hadu a cikin farji ya ƙare a SSTs2.A cikin insemination na kajin wucin gadi a cikin farji, adadin spermatozoa ya kai SST yana nuna karuwa 24 hours bayan balaga.Ya zuwa yanzu, tsarin zaɓin maniyyi yayin wannan tsari ba a bayyana ba, kuma motsin maniyyi na iya taka muhimmiyar rawa wajen ɗaukar maniyyi SST.Saboda kauri da kaurin bangon bututun fallopian, yana da wuya a iya lura da motsin maniyyi kai tsaye a cikin bututun tsuntsaye na fallopian.Saboda haka, ba mu da ainihin ilimin yadda maniyyi ke canzawa zuwa SST bayan hadi.
Kwanan nan an gane Rheology a matsayin muhimmin abu mai sarrafa jigilar maniyyi a cikin al'aurar mammalian.Dangane da iyawar maniyyi masu motsi don yin ƙaura ba tare da bata lokaci ba, Zaferani et al8 sun yi amfani da tsarin corra microfluidic don ware maniyyi mai motsi daga samfuran maniyyi da aka rubuta.Irin wannan nau'in rarraba maniyyi yana da mahimmanci don maganin rashin haihuwa na likita da bincike na asibiti, kuma an fifita shi fiye da hanyoyin gargajiya waɗanda suke da lokaci da aiki mai tsanani kuma suna iya yin tasiri ga tsarin halittar maniyyi da daidaiton tsari.Sai dai har ya zuwa yau, ba a gudanar da bincike kan illar fitar da kaji ke haifar da motsin maniyyi ba.
Ko da kuwa tsarin da ke kula da maniyyi da aka adana a cikin SST, masu bincike da yawa sun lura cewa mazaunin spermatozoa agglutinate kai-da-kai a cikin SST na kaji 9, 10, quails 2, da turkeys 11 don samar da gunkin maniyyi agglutinated.Mawallafa sun ba da shawarar cewa akwai hanyar haɗi tsakanin wannan agglutination da kuma adana dogon lokaci na spermatozoa a cikin SST.
Tingari da Lake12 sun ba da rahoton wata alaƙa mai ƙarfi tsakanin spermatozoa a cikin gland mai karɓar maniyyi na kaza kuma sun yi tambaya ko avian spermatozoa agglutinate daidai da mammalian spermatozoa.Sun yi imanin cewa haɗin kai mai zurfi tsakanin maniyyi a cikin vas deferens na iya zama saboda damuwa da kasancewar yawan adadin maniyyi a cikin karamin wuri.
Lokacin da ake kimanta halayen spermatozoa akan sabon rataye gilashin nunin faifai, ana iya ganin alamun tashin hankali na wucin gadi, musamman a gefuna na ɗigon ruwan maniyyi.Duk da haka, agglutination sau da yawa yakan damu da aikin jujjuyawar da ke hade da ci gaba da motsi, wanda ke bayyana yanayin wucin gadi na wannan sabon abu.Masu binciken sun kuma lura cewa lokacin da aka ƙara diluent a cikin maniyyi, tarin tantanin halitta "zari-kamar" sun bayyana.
An yi ƙoƙari na farko don kwaikwayon maniyyi ta hanyar cire wata siririyar waya daga digon rataye, wanda ya haifar da wani tsayin daka mai kama da maniyyi yana fitowa daga digon maniyyi.Nan da nan spermatozoa ta jera layi ɗaya a cikin vesicle, amma gaba ɗaya rukunin ya ɓace da sauri saboda iyakancewar 3D.Don haka, don nazarin spermatozoa agglutination, ya zama dole a lura da motsi da halayen spermatozoa kai tsaye a cikin keɓaɓɓen tubules na ajiyar maniyyi, wanda ke da wahala a cimma.Sabili da haka, ya zama dole don haɓaka kayan aiki wanda ke kwaikwayon spermatozoa don tallafawa nazarin motsin maniyyi da halayen agglutination.Brillard et al13 sun ruwaito cewa matsakaicin tsayin tubules na maniyyi a cikin kajin manya shine 400-600 µm, amma wasu SSTs na iya zama tsayin 2000 µm.Mero da Ogasawara14 sun raba glandan seminiferous zuwa manyan tubules na maniyyi masu girma da marasa girma, dukansu iri ɗaya ne tsawon (~ 500 µm) da faɗin wuyansa (~ 38 µm), amma ma'anar lumen diamita na tubules shine 56.6 da 56.6 µm.., bi da bi 11.2 μm, bi da bi.A cikin binciken na yanzu, mun yi amfani da na'urar microfluidic tare da girman tashoshi 200 µm × 20 µm (W × H), wanda sashin giciye ya ɗan kusa da na SST.Bugu da kari, mun yi nazari kan motsin maniyyi da halayyar agglutination a cikin ruwa mai gudana, wanda ya yi daidai da hasashen Foreman cewa ruwan da SST epithelial sel ke samarwa yana kiyaye maniyyi a cikin lumen a cikin alkibla (rheological).
Manufar wannan binciken shine don shawo kan matsalolin lura da motsin maniyyi a cikin bututun fallopian da kuma guje wa matsalolin nazarin ilimin rheology da halayyar spermatozoa a cikin yanayi mai ƙarfi.An yi amfani da na'urar microfluidic wanda ke haifar da matsin lamba don daidaita motsin maniyyi a cikin al'aurar kaza.
Lokacin da aka ɗora diluted samfurin maniyyi (1:40) a cikin na'urar microchannel, ana iya gano nau'ikan motsin maniyyi guda biyu (waɗanda aka keɓe da kuma ɗaure maniyyi).Bugu da ƙari, spermatozoa yana kula da yin iyo a kan halin yanzu (mai kyau rheology; bidiyo 1, 2). Kodayake tarin maniyyi yana da ƙananan saurin gudu fiye da na maniyyi kaɗai (p <0.001), sun ƙara yawan adadin maniyyi da ke nuna rheotaxis mai kyau (p <0.001; Table 2). Kodayake tarin maniyyi yana da ƙananan saurin gudu fiye da na maniyyi kaɗai (p <0.001), sun ƙara yawan adadin maniyyi da ke nuna rheotaxis mai kyau (p <0.001; Table 2). Хотя пучки сперматозоидов имели более низкую скорость. ерматозоидов, демонстрирующих положительный реотаксис (p <0,001; таблица 2). Kodayake spermatozoa bundles suna da ƙananan sauri fiye da na spermatozoa guda ɗaya (p <0.001), sun ƙara yawan adadin spermatozoa yana nuna rheotaxis mai kyau (p <0.001; Table 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显示阳性显示阳性孤的显示阳性孤示阳<0.001; 2）尽管 精子束 的 速度 低于 孤独 的 速度 （p <0.001） (p <0.001; 2….)))) Хотя скорость пучков сперматозоидов была ниже, чем у одиночныh дов с положительной реологией (p <0,001; таблица 2). Kodayake saurin ɗigon maniyyi ya kasance ƙasa da na spermatozoa guda ɗaya (p <0.001), sun ƙara yawan adadin spermatozoa tare da ingantaccen rheology (p <0.001; Table 2).Kyakkyawan rheology na spermatozoa guda ɗaya da tufts an kiyasta kusan 53% da 85%, bi da bi.
An lura cewa spermatozoa na kajin sharkasi nan da nan bayan fitar maniyyi ya zama nau'i na layi, wanda ya ƙunshi mutane da dama.Wadannan tufts suna ƙaruwa da tsayi da kauri akan lokaci kuma suna iya kasancewa a cikin vitro na sa'o'i da yawa kafin su rabu (bidiyo 3).Wadannan dauren filamentous suna da siffa kamar echidna spermatozoa wanda ke samuwa a ƙarshen epididymis.An gano maniyyin kaza na Sharkashi yana da babban hali don yin agglutinate da samar da tarin reticulate a cikin ƙasa da minti ɗaya bayan tattarawa.Waɗannan katako suna da ƙarfi kuma suna iya mannewa ga kowane bango na kusa ko a tsaye.Ko da yake ɗigon maniyyi yana rage saurin ƙwayoyin maniyyi, a bayyane yake cewa macroscopically suna haɓaka layinsu.Tsawon ƙullun ya bambanta dangane da adadin maniyyi da aka tattara a daure.An keɓance sassa biyu na dam ɗin: ɓangaren farko, gami da shugaban kyauta na maniyyi agglutinated, da ɓangaren ƙarshen, gami da wutsiya da ƙarshen ƙarshen maniyyi.Yin amfani da kyamara mai sauri (950fps), an lura da shugabannin kyauta na agglutinated spermatozoa a cikin farkon ɓangaren ƙwayar cuta, da alhakin motsin daɗaɗɗen saboda motsin motsin su, yana jawo sauran zuwa cikin damfara tare da motsi na helical (Video 4).Duk da haka, a cikin dogon tufts, an lura cewa wasu kawunan maniyyi masu kyauta suna mannewa ga jiki kuma sashin da ya dace na tuft yana aiki azaman vanes don taimakawa tudun tudun.
Yayin da ruwa ke tafiya a hankali, daurin maniyyi suna tafiya daidai da juna, duk da haka, sai su fara yin rufa-rufa suna manne da duk abin da yake har yanzu, don gudun kada ruwan na yanzu ya wanke shi yayin da saurin gudu ya karu.Dauren suna samuwa ne a lokacin da ɗimbin ɗimbin ƙwayoyin maniyyi suka kusanci juna, sai su fara motsawa tare da naɗa juna, sannan su manne da wani abu mai ɗaci.Hoto na 1 da na 2 sun nuna yadda maniyyi ke kusanto juna, inda suka zama mahaɗa yayin da wutsiyoyi ke zagaye da juna.
Masu binciken sun yi amfani da matsa lamba na hydrostatic don haifar da kwararar ruwa a cikin microchannel don nazarin rheology na maniyyi.An yi amfani da microchannel mai girman 200 µm × 20 µm (W × H) da tsawon 3.6 µm.Yi amfani da ƙananan tashoshi tsakanin kwantena tare da sirinji masu dacewa a ƙarshen.An yi amfani da canza launin abinci don sanya tashoshi mafi bayyane.
Daura igiyoyi masu haɗin haɗin kai da na'urorin haɗi zuwa bango.An ɗauki bidiyon tare da maƙalli mai ban mamaki na lokaci.Tare da kowane hoto, ana gabatar da bambance-bambancen microscope da hotunan taswira.(A) Haɗin kai tsakanin koguna guda biyu yana tsayayya da kwarara saboda motsi na helical (kibiya ja).(B) Haɗin kai tsakanin bututun bututu da bangon tashar (kibiyoyin ja), a lokaci guda an haɗa su da wasu nau'ikan guda biyu (kibiyoyin rawaya).(C) Maniyyi daure a cikin microfluidic tashar fara haɗi da juna (ja kibiyoyi), forming raga na maniyyi daure.(D) Samar da hanyar sadarwa na daure maniyyi.
Lokacin da aka ɗora diluted na maniyyi a cikin na'urar microfluidic kuma an halicci kwarara, an lura da katakon maniyyi yana motsawa a kan hanyar da ke gudana.Ƙunƙarar ta dace da ganuwar microchannels, kuma kawuna masu kyauta a cikin ɓangaren farko na ƙullun sun dace da su (bidiyo 5).Har ila yau, suna manne wa kowane ɓangarorin da ke tsaye a hanyarsu, kamar tarkace, don ƙin yarda da abin da yanzu ke ɗauke da shi.A tsawon lokaci, waɗannan tufts sun zama dogayen filaments masu kama wasu maniyyi guda ɗaya da guntun tufts (Video 6).Yayin da magudanar ya fara raguwa, dogayen layin maniyyi sun fara samar da hanyar sadarwa na layin maniyyi (Bidiyo 7; Hoto na 2).
A babban saurin kwarara (V> 33 µm/s), motsin zaren zaren yana ƙaruwa azaman yunƙurin kama maniyyi da yawa waɗanda ke samar da daure mafi kyawu don tsayayya da ƙarfin kwararar. A babban saurin kwarara (V> 33 µm/s), motsin zaren zaren yana ƙaruwa azaman yunƙurin kama maniyyi da yawa waɗanda ke samar da daure mafi kyawu don tsayayya da ƙarfin kwararar. При высокой скорости потока (V > 33 мкм/с) спиралевидные тдельных сперматозоидов, образующих пучки, которые лучше. A babban adadin kwarara (V> 33 µm/s), motsin motsi na igiyoyi yana ƙaruwa yayin da suke ƙoƙarin kama maniyyi da yawa waɗanda ke samar da daure waɗanda suka fi dacewa da tsayayyar ƙarfin kwararar.在高流过流动的漂移力。在 高 流速 (v> 33 µm/s) 时 , 的 螺旋 展开的 漂移力............... При высоких скоростях потока (V > 33 мкм/с) спиральное движение матозоидов, образующих пучки, чтобы лучше сопротивляться силам дрейфа потока. A babban adadin kwarara (V> 33 µm/s), motsi na filament yana ƙaruwa a cikin yunƙurin kama maniyyi da yawa waɗanda ke samar da daure don mafi kyawun jure wa rundunonin ruwa.Sun kuma yi ƙoƙarin haɗa microchannels zuwa bangon gefe.
An gano dauren maniyyi a matsayin gungu na kawunan maniyyi da wutsiyoyi masu murzawa ta hanyar amfani da hasken microscopy (LM).Har ila yau, an gano tarin maniyyi masu tarin tarin yawa a matsayin karkatattun kawuna da tarin tuta, wutsiyar maniyyi da yawa, kawunan maniyyi da ke manne da wutsiya, da kan maniyyi masu lankwashe tsakiya a matsayin dunkule masu hade da yawa.microscope na watsawa (TEM).Na'urar bincike ta microscopy (SEM) ta nuna cewa tarin maniyyin sun kasance da tarin kawunan maniyyi kuma tarin maniyyi ya nuna hanyar sadarwa na nannade.
An yi nazarin ilimin halittar jiki da ultrastructure na spermatozoa, da samuwar spermatozoa daure ta amfani da haske microscopy (rabin sashe), scanning electron microscopy (SEM) da kuma watsa electron microscopy (TEM), sperm smears aka stained da acridine orange da kuma bincika ta amfani da epifluorescence microscopy.
Smear smear tare da acridine orange (Fig. 3B) ya nuna cewa kawunan maniyyi sun makale tare kuma an rufe su da kayan asiri, wanda ya haifar da samuwar manyan tufts (Fig. 3D).Rukunin maniyyi sun ƙunshi tarin maniyyi tare da hanyar sadarwa na wutsiyoyi (Fig. 4A-C).Dauren maniyyi sun ƙunshi wutsiyoyi da yawa na maniyyi masu makale tare (Fig. 4D).Asirin (Fig. 4E,F) ya rufe kawunan maniyyi daurin.
Samar da tarin spermatozoa Yin amfani da yanayin bambance-bambancen microscopy da sperm smears wanda aka yi da acridine orange, ya nuna cewa kawunan spermatozoa yana tare.(A) Farkon samuwar maniyyi yana farawa da maniyyi (fararen da'irar) da maniyyi uku (rawaya da'ira), tare da karkace daga wutsiya kuma yana ƙarewa a kai.(B) Hoton faifan maniyyi mai tabo da lemu na acridine yana nuna madaidaicin kawunan maniyyi (kibau).Fitowar ta rufe kai (s).Girman girma × 1000. (C) Haɓaka babban katako da aka ɗauka ta hanyar kwarara a cikin tashar microfluidic (ta yin amfani da kyamara mai sauri a 950 fps).(D) Micrograph na smear maniyyi da aka tabo da lemu na acridine yana nuna manyan tufa (kibiyoyi).Girma: ×200.
Ana duba micrograph na lantarki na katakon maniyyi da maniyyi mai tabo da orange acridine.(A, B, D, E) su ne dijital launi na sikanin lantarki micrographs na spermatozoa, da kuma C da F su ne micrographs na acridine orange tabo smears nuna abin da aka makala na mahara spermatozoa kunshe da caudal yanar gizo.(AC) Ana nuna tarin maniyyi azaman hanyar sadarwa na wutsiyoyi (kibiyoyi).(D) Manne da yawa maniyyi (tare da m abu, ruwan hoda shaci, kibiya) nada a kusa da wutsiya.E da Fspermatozoa sun kafa daure tare da sifofi masu kama da vortex da yawa (F).(C) × 400 da (F) × 200 girma.
Yin amfani da microscope na lantarki watsawa, mun gano cewa ɗigon maniyyi sun haɗa wutsiyoyi (Fig. 6A, C), kawunan da aka haɗe zuwa wutsiya (Fig. 6B), ko kawunan da aka haɗe zuwa wutsiya (Fig. 6D).Shugabannin spermatozoa a cikin dunƙule suna lanƙwasa, suna nunawa a cikin sashe na yankuna biyu na nukiliya (Fig. 6D).A cikin ƙunƙun ɓarna, spermatozoa yana da murɗaɗɗen kai mai yankuna biyu na nukiliya da yankuna masu yawa (Fig. 5A).
Micrograph mai launi mai launi na dijital yana nuna wutsiyoyi masu haɗawa a cikin tarin maniyyi da kayan haɓakawa da ke haɗa kawunan maniyyi.(A) Haɗe wutsiya na adadi mai yawa na spermatozoa.Yi la'akari da yadda wutsiya ke kallon duka hotuna (kibiya) da kuma shimfidar wuri (kibiya).(B) Kan (kibiya) na maniyyi yana haɗe da wutsiya (kibiya).(C) An haɗe wutsiyoyi da yawa (kibiyoyi).(D) Abubuwan Agglutination (AS, blue) suna haɗa kawunan maniyyi guda huɗu (purple).
An yi amfani da microscopy na lantarki don gano kawunan maniyyi a cikin ɗimbin maniyyi da aka rufe da ɓoye ko membranes (Hoto na 6B), wanda ke nuni da cewa daurin maniyyi an haɗa su ta wani abu na waje.Abubuwan agglutinated an tattara su a cikin maniyyi (jellyfish head-like meeting; Fig. 5B) kuma ya faɗaɗa nisa, yana ba da bayyanar rawaya mai haske a ƙarƙashin microscopy mai haske lokacin da aka lalata shi da acridine orange (Fig. 6C).Wannan abu a bayyane yake a bayyane a ƙarƙashin na'urar dubawa kuma ana ɗaukarsa a matsayin mai ɗaure.Sassan ɓangarorin da ba su da bakin ciki (Fig. 5C) da smears ɗin maniyyi da aka tabo da lemu na acridine sun nuna ɗimbin maniyyi masu ɗauke da kawuna masu yawa da wutsiyoyi masu murɗa (Fig. 5D).
Hotunan hoto daban-daban suna nuna tarin kawunan maniyyi da wutsiyoyi masu ninke ta amfani da hanyoyi daban-daban.(A) Mikrograph mai launi na dijital da ke jujjuya nau'ikan nau'ikan nau'ikan nau'ikan nau'ikan nau'ikan maniyyi da ke nuna kan maniyyi mai murɗa tare da tsakiya mai kashi biyu (blue) da ɓangarori da yawa na flagellar (kore).(B) Micrograph na lantarki mai launi na dijital yana nuna gungu na kawunan maniyyi masu kama da jellyfish (kiban) waɗanda da alama an rufe su.(C) Sashi mai sirara yana nuna kanun maniyyi da aka tara (kibiyoyi) da wutsiyoyi masu lanƙwasa (kibiyoyi).(D) Micrograph na smear maniyyi wanda aka tabo da lemu acridine yana nuna tarin kawunan maniyyi (kibiyoyi) da wutsiyoyi masu lanƙwasa (kibiyoyi).Lura cewa wani abu mai ɗaki (S) yana rufe kan maniyyi.(D) × 1000 girma.
Yin amfani da microscope na lantarki watsawa (Fig. 7A), an kuma lura cewa kawunan maniyyi sun karkace kuma tsakiya suna da siffar karkace, kamar yadda aka tabbatar ta hanyar sperm smears da aka yi da acridine orange kuma an yi nazari ta hanyar amfani da microscopy na fluorescence (Fig. 7B).
(A) Na'urar watsa launi na dijital micrograph da (B) Acridine orange mai tabo smear yana nuna kawuna da aka haɗe da kawunan maniyyi da wutsiyoyi (kibiyoyi).(B) × 1000 girma.
Wani bincike mai ban sha'awa shi ne cewa maniyyi Sharkazi ya haɗu ya zama nau'in filamentous na wayar hannu.Kaddarorin waɗannan daure suna ba mu damar fahimtar yiwuwar rawar da suke takawa a cikin sha da adana spermatozoa a cikin SST.
Bayan jima'i, maniyyi ya shiga cikin farji kuma ya sha wani tsari mai tsanani, wanda ya haifar da iyakacin adadin maniyyi ya shiga SST15,16.Har zuwa yau, hanyoyin da maniyyi ke shiga da fita daga SST ba su da tabbas.A cikin kiwon kaji, ana adana spermatozoa a cikin SST na tsawon makonni 2 zuwa 10, dangane da nau'in6.Rikici ya kasance game da yanayin maniyyi yayin ajiya a cikin SST.Shin suna motsi ne ko kuma suna hutawa?A wasu kalmomi, ta yaya ƙwayoyin maniyyi ke kula da matsayinsu a cikin SST na tsawon lokaci?
Forman4 ya ba da shawarar cewa za a iya bayyana wurin zama da SST ta hanyar motsin maniyyi.Marubutan sun yi hasashen cewa maniyyi yana kula da matsayinsa ta hanyar yin iyo a kan magudanar ruwa da SST epithelium ya haifar kuma ana fitar da maniyyi daga SST lokacin da saurinsu ya fadi kasa da inda suke fara komawa baya saboda rashin kuzari.Zaniboni5 ya tabbatar da kasancewar aquaporins 2, 3, da 9 a cikin sashin apical na sel epithelial SST, wanda zai iya tallafawa tsarin ajiyar maniyyi na Foreman a kaikaice.A cikin binciken da muke yi yanzu, mun gano cewa kusan rabin maniyyi na Sharkashi yana nuna ingantaccen rheology a cikin ruwan da ke gudana, kuma adadin maniyyi da aka samu yana kara yawan maniyyin da ke nuna rheology mai kyau, kodayake agglutination yana raguwa.Yadda ƙwayoyin maniyyi ke tafiya sama da bututun fallopian na tsuntsu zuwa wurin da ake hadi ba a cika fahimta ba.A cikin dabbobi masu shayarwa, ruwan follicular chemo yana jan hankalin spermatozoa.Duk da haka, chemoattractants an yi imani da kai tsaye spermatozoa zuwa kusanci mai nisa7.Saboda haka, wasu hanyoyin suna da alhakin jigilar maniyi.Ƙarfin maniyyi don daidaitawa da gudana a kan ruwan bututun fallopian da aka saki bayan jima'i an ruwaito shi ne babban abin da ke haifar da niyya ga maniyyi a cikin mice.Parker 17 ya ba da shawarar cewa spermatozoa ƙetare oviducts ta yin iyo a kan ciliary halin yanzu a cikin tsuntsaye da dabbobi masu rarrafe.Ko da yake ba a gwada shi a cikin tsuntsaye ba, Adolphi18 shine farkon wanda ya gano cewa maniyyi na avian yana ba da sakamako mai kyau lokacin da aka ƙirƙiri wani ɗan ƙaramin ruwa tsakanin murfin murfin da zamewa tare da tsiri na takarda.Rheology.Hino da Yanagimachi [19] sun sanya wani hadadden ovary-tubal-uterine na linzamin kwamfuta a cikin zobe na turare kuma suka allurar 1 µl na tawada a cikin isthmus don ganin yadda ruwa ke gudana a cikin bututun fallopian.Sun lura da motsin ƙanƙancewa da annashuwa a cikin bututun fallopian, inda duk ƙwallan tawada ke motsawa a hankali zuwa ampulla na bututun fallopian.Marubutan sun jaddada mahimmancin kwararar ruwan tubal daga kasa zuwa manyan bututun fallopian na sama don tayar da maniyyi da hadi.Brillard20 ta ruwaito cewa a cikin kaji da turkeys, spermatozoa na yin ƙaura ta hanyar motsa jiki daga ƙofar farji, inda aka adana su, zuwa mahadar mahaifa-farji, inda ake adana su.Koyaya, ba a buƙatar wannan motsi tsakanin mahaɗar uterovaginal da infundibulum saboda ana jigilar maniyyi ta hanyar ƙaura.Sanin waɗannan shawarwarin da suka gabata da sakamakon da aka samu a cikin binciken na yanzu, ana iya ɗauka cewa ikon spermatozoa don motsawa sama (rheology) yana daya daga cikin kaddarorin da tsarin zaɓin ya dogara.Wannan yana ƙayyade yadda spermatozoa ke wucewa ta cikin farji da shigar su cikin CCT don ajiya.Kamar yadda Forman4 ya ba da shawara, wannan na iya sauƙaƙe hanyar shigar da maniyyi shiga SST da mazauninsa na wani ɗan lokaci sannan kuma ya fita lokacin da saurin su ya fara raguwa.
A gefe guda, Matsuzaki da Sasanami 21 sun ba da shawarar cewa spermatozoa avian suna fuskantar canje-canjen motsi daga barci zuwa motsi a cikin sassan haihuwa na maza da mata.An ba da shawarar hana motsin motsin maniyyi a cikin SST don bayyana tsawon lokacin ajiyar maniyyi sannan kuma sake farfadowa bayan barin SST.A ƙarƙashin yanayin hypoxic, Matsuzaki et al.1 ya ruwaito babban samarwa da sakin lactate a cikin SST, wanda zai iya haifar da hana motsin maniyyi mazauna.A wannan yanayin, mahimmancin rheology na maniyyi yana nunawa a cikin zaɓi da kuma sha na spermatozoa, kuma ba a cikin ajiyar su ba.
Maniyyi agglutination juna ana la'akari da wani m bayani ga dogon ajiya lokaci na maniyyi a cikin SST, domin wannan shi ne na kowa tsari na sperm rike a cikin kaji2,22,23.Bakst et al.2 ya lura cewa mafi yawan spermatozoa suna manne da juna, suna samar da tarawar fascicular, da kuma spermatozoa guda ɗaya da wuya a samu a cikin quail CCM.A gefe guda, Wen et al.24 an lura da ƙarin warwatse spermatozoa da ƙarancin spermatozoa tufts a cikin SST lumen a cikin kaji.Dangane da waɗannan abubuwan lura, ana iya ɗauka cewa haɓakar haɓakar maniyyi ya bambanta tsakanin tsuntsaye da tsakanin spermatozoa a cikin maniyyi ɗaya.Bugu da kari, Van Krey et al.9 ya ba da shawarar cewa bazuwar bazuwar maniyyi na agglutinated shine ke da alhakin shigar da maniyyi a hankali a cikin lumen na bututun fallopian.Bisa ga wannan hasashe, spermatozoa tare da ƙananan ƙarfin agglutination ya kamata a fitar da shi daga SST da farko.A cikin wannan mahallin, ikon spermatozoa zuwa agglutinate na iya zama abin da ke tasiri sakamakon gasar maniyyi a cikin tsuntsaye masu datti.Bugu da ƙari, tsawon lokacin da maniyyi agglutinated ya rabu, ana kiyaye tsawon lokacin haihuwa.
Ko da yake an lura da tarawar spermatozoa da tarawa cikin damfara a cikin bincike da yawa2,22,24, ba a bayyana su dalla-dalla ba saboda sarƙaƙƙiyar kallon kinematic ɗin su a cikin SST.An yi yunƙuri da yawa don nazarin sperm agglutination a cikin vitro.An lura da haɗuwa mai yawa amma na wucin gadi lokacin da aka cire siririyar waya daga digon iri.Wannan yana haifar da gaskiyar cewa kumfa mai elongated yana fitowa daga digo, yana kwaikwayon glandar seminal.Saboda iyakoki na 3D da gajerun lokutan bushewar drip, duk toshewar cikin sauri ya faɗi cikin lalacewa9.A cikin binciken na yanzu, ta yin amfani da kajin Sharkashi da kwakwalwan kwamfuta na microfluidic, mun sami damar bayyana yadda waɗannan tufts ke samuwa da yadda suke motsawa.Maniyyi daurin kafa nan da nan bayan tarin maniyyi kuma an same shi yana motsawa a cikin karkace, yana nuna ingantaccen rheology lokacin da yake cikin kwarara.Bugu da ƙari kuma, lokacin da aka kalli macroscopically, an lura da tarin maniyyi don ƙara layin motsi idan aka kwatanta da keɓewar spermatozoa.Wannan yana nuna cewa haɓakar maniyyi na iya faruwa kafin shigar SST kuma samar da maniyyi bai iyakance ga ƙaramin yanki ba saboda damuwa kamar yadda aka nuna a baya (Tingari da Lake12).A lokacin samuwar spermatozoa, spermatozoa na yin iyo a cikin synchrony har sai sun zama mahaɗa, sai wutsiyarsu ta naɗe juna kuma kan naman maniyyi ya kasance kyauta, amma wutsiya da nisa na maniyyi suna tsayawa tare da wani abu mai danko.Sabili da haka, kai mai kyauta na ligament yana da alhakin motsi, yana jan sauran ligament.Binciken da aka yi da na’urorin lantarki na maniyyin maniyyi ya nuna kawunan maniyyi da aka makala da abubuwa masu dunkulewa, wanda hakan ke nuni da cewa an makala kawunan maniyyin a cikin daurin hutawa, wanda watakila ya faru ne bayan isa wurin ajiya (SST).
Lokacin da sperm smear ya zama tabo da acridine orange, extracellular abu a kusa da maniyyi sel za a iya gani a karkashin wani mai kyalli microscope.Wannan sinadari yana ba da damar ɗimbin maniyyi don mannewa da mannewa ga duk wani abu da ke kewaye da shi don kada su yi tafiya tare da kewaye.Don haka, abubuwan da muka lura sun nuna rawar da mannewar spermatozoa ke yi a cikin nau'in dam na wayar hannu.Ƙarfinsu na yin iyo a kan halin yanzu da kuma manne wa saman da ke kusa yana ba da damar maniyyi ya dade a cikin SST.
Rothschild25 ya yi amfani da kyamarar haemocytometry don nazarin rarrabawar ruwan maniyyi na bovine a cikin digo na dakatarwa, yana ɗaukar hotuna ta hanyar kyamara tare da axis na gani a tsaye da a kwance na microscope.Sakamakon ya nuna cewa spermatozoa ya jawo hankalin saman ɗakin.Mawallafa sun ba da shawarar cewa za a iya samun hulɗar hydrodynamic tsakanin maniyyi da saman.Idan aka yi la’akari da haka, tare da iyawar maniyyin kajin Sharkashi don samar da tutsu masu ɗanɗano, zai iya ƙara yuwuwar cewa maniyyi ya manne da bangon SST kuma a adana shi na dogon lokaci.
Bccetti da Afzeliu26 sun ruwaito cewa ana buƙatar sperm glycocalyx don gane gamete da agglutination.Forman10 ya lura cewa hydrolysis na α-glycosidic bonds a cikin glycoprotein-glycolipid coatings ta hanyar kula da maniyyi avian tare da neuraminidase ya haifar da raguwar haihuwa ba tare da rinjayar motsin maniyyi ba.Mawallafa sun ba da shawarar cewa tasirin neuraminidase akan glycocalyx yana lalata maniyyin maniyyi a mahadar mahaifa-farji, don haka rage yawan haihuwa.Abubuwan da suka lura ba za su iya watsi da yiwuwar cewa maganin neuraminidase zai iya rage maniyyi da kuma oocyte ganewa.Forman da Engel10 sun gano cewa haifuwa ya ragu lokacin da aka yi wa kaji yaduwa ta cikin farji tare da maniyyi da aka yi da neuraminidase.Duk da haka, IVF tare da neuraminidase da aka kula da maniyyi bai shafi haihuwa ba idan aka kwatanta da kaji.Marubutan sun kammala da cewa canje-canjen da ake samu a cikin glycoprotein-glycolipid shafi a kusa da maniyyi na maniyyi ya rage karfin maniyyi yin takin ta hanyar tauye maniyyi a mahadar mahaifa da farji, wanda hakan ya kara hasarar maniyyi saboda gudun haduwar mahaifa-farji, amma ba ya shafar maniyyi da kwai.
A cikin turkeys Bakst da Bauchan 11 sun sami ƙananan vesicles da ɓawon burodi a cikin lumen na SST kuma sun lura cewa wasu daga cikin waɗannan granules sun haɗu da ƙwayar maniyyi.Mawallafa sun ba da shawarar cewa waɗannan alaƙa na iya taimakawa wajen adana dogon lokaci na spermatozoa a cikin SST.Sai dai masu binciken ba su fayyace tushen wadannan barbashi ba, ko dai kwayoyin epithelial na CCT ne suka kebe su, ko kuma tsarin haihuwa na namiji ne suka samar da su, ko kuma maniyyin da kansa ya samar.Hakanan, waɗannan ƙwayoyin suna da alhakin agglutination.Grützner et al27 ya ruwaito cewa ƙwayoyin epididymal epithelial suna samar da kuma ɓoye wani takamaiman furotin wanda ake buƙata don samar da sassan seminal na pore guda ɗaya.Har ila yau, marubutan sun ba da rahoton cewa tarwatsa waɗannan daure ya dogara da hulɗar sunadaran epididymal.Nixon et al28 sun gano cewa adnexa yana ɓoye furotin, acidic cysteine-rich osteonectin;SPARC tana shiga cikin samuwar tudun maniyyi a cikin gajeriyar echidnas da platypuses.Watsewar waɗannan katako yana da alaƙa da asarar wannan furotin.
A cikin binciken na yanzu, bincike na ultrastructural ta amfani da microscopy na lantarki ya nuna cewa spermatozoa yana manne da adadi mai yawa.Ana tsammanin waɗannan abubuwa suna da alhakin haɓakar haɓakar da ke tattare da juna tsakanin da kuma kewayen kawunan da ke gaba, amma a ƙananan ƙima a cikin yankin wutsiya.Muna ɗauka cewa wannan abu mai banƙyama yana fitar da shi daga tsarin haihuwa na namiji (epididymis ko vas deferens) tare da maniyyi, tun da yake sau da yawa muna ganin maniyyi yana rabuwa da lymph da plasma na jini yayin fitar maniyyi.An ba da rahoton cewa yayin da spermatozoa avian ke wucewa ta cikin epididymis da vas deferens, suna fuskantar canje-canje masu alaka da balagagge wanda ke tallafawa ikon su na ɗaure sunadaran da kuma samun glycoproteins masu alaƙa da plasma lemma.Dagewar wadannan sunadaran a kan mazaunin maniyyi a cikin SST yana nuna cewa waɗannan sunadaran zasu iya rinjayar samun kwanciyar hankali na maniyyi 30 da kuma ƙayyade yawan haihuwa 31.Ahammad et al32 ya ruwaito cewa spermatozoa da aka samu daga sassa daban-daban na tsarin haihuwa na maza (daga testes zuwa distal vas deferens) ya nuna karuwar ci gaba a cikin iya aiki a karkashin yanayin ajiyar ruwa, ba tare da la'akari da yawan zafin jiki ba, kuma yiwuwar a cikin kaji yana karuwa a cikin tubes na fallopian bayan haihuwa na wucin gadi.
Sharkashi kaji tufts yana da halaye da ayyuka daban-daban fiye da sauran nau'ikan kamar echidnas, platypuses, mice na itace, berayen barewa, da aladun Guinea.A cikin kajin sharkasi, samuwar maniyyi daure ya rage saurin ninka idan aka kwatanta da maniyyi daya.Duk da haka, waɗannan daure sun ƙara yawan rheologically tabbatacce spermatozoa da kuma ƙara ikon spermatozoa don daidaita kansu a cikin wani tsauri yanayi.Don haka, sakamakonmu ya tabbatar da shawarar da ta gabata cewa haɓakar maniyyi a cikin SST yana da alaƙa da ajiyar maniyyi na dogon lokaci.Mun kuma yi hasashe cewa kwarjinin maniyyi don samar da tufts na iya sarrafa adadin asarar maniyyi a cikin SST, wanda zai iya canza sakamakon gasar maniyyi.Bisa ga wannan zato, spermatozoa tare da ƙananan ƙarfin agglutination saki SST da farko, yayin da spermatozoa tare da babban ƙarfin agglutination yana samar da mafi yawan zuriya.Samar da tarin maniyyi guda-pore yana da amfani kuma yana shafar rabon iyaye da yara, amma yana amfani da wata hanya ta daban.A cikin echidnas da platypuses, spermatozoa an shirya su a layi daya da juna don ƙara saurin ci gaba na katako.Dauren echidnas suna motsawa kusan sau uku da sauri fiye da maniyyi ɗaya.An yi imani da cewa samuwar irin wannan maniyyi tufts a echidnas ne wani juyin halitta karbuwa don ci gaba da rinjaye, tun da mata ne m kuma yawanci saduwa da maza da yawa.Don haka, maniyyi daga maniyyi daban-daban suna fafatawa sosai don haɗewar kwai.
Agglutinated spermatozoa na sharkasi kaji yana da sauƙin gani ta amfani da microscopy bambanci na lokaci, wanda ake ganin yana da fa'ida saboda yana ba da damar yin nazari cikin sauƙi na halayen spermatozoa a cikin vitro.Hanyar da samuwar maniyyi ke haifar da haifuwa a cikin kajin sharkasi shi ma ya sha bamban da yadda ake gani a wasu dabbobi masu shayarwa dake wakiltar halayen maniyyin hadin gwiwa irin su berayen itace, inda wasu maniyyin maniyyi ya kai ga kwai, yana taimakawa wasu masu alaka da su kai ga lalata kwai.don tabbatar da kanku.halin altruistic.Haɗuwa da kai 34. Wani misali na halayyar haɗin kai a cikin maniyyi an gano shi a cikin berayen barewa, inda spermatozoa ya sami damar ganowa da haɗuwa tare da mafi yawan jinsin maniyyi da kuma samar da ƙungiyoyin haɗin gwiwa don ƙara saurin su idan aka kwatanta da spermatozoa35 maras dangantaka.
Sakamakon da aka samu a cikin wannan binciken bai saba wa ka'idar Foman na adana dogon lokaci na spermatozoa a cikin SWS ba.Masu binciken sun bayar da rahoton cewa, kwayoyin halittar maniyyi suna ci gaba da tafiya a cikin magudanar kwayoyin epithelial da ke rufe SST na wani lokaci mai tsawo, kuma bayan wani lokaci, ma'ajin makamashi na kwayoyin halittar maniyyi sun lalace, wanda ke haifar da raguwar saurin gudu, wanda ke ba da damar fitar da kananan abubuwa masu nauyin kwayoyin.makamashi na spermatozoa tare da kwararar ruwa daga lumen na SST Ramin tube na fallopian.A cikin binciken na yanzu, mun lura cewa rabin maniyyi daya ya nuna ikon yin iyo a kan ruwan da ke gudana, kuma mannewar su a cikin damfara yana kara karfin su na nuna rheology mai kyau.Bugu da ƙari kuma, bayananmu sun yi daidai da na Matsuzaki et al.1 wanda ya ba da rahoton cewa ƙara yawan ƙwayar lactate a cikin SST na iya hana motsin maniyyi mazaunin.Koyaya, sakamakonmu yana bayyana samuwar ligaments masu motsi na maniyyi da halayen rheological a gaban yanayi mai ƙarfi a cikin microchannel a ƙoƙarin bayyana halayen su a cikin SST.Bincike na gaba na iya mayar da hankali kan tantance abubuwan da ke tattare da sinadarai da asalin wakili na agglutinating, wanda babu shakka zai taimaka wa masu bincike haɓaka sabbin hanyoyin adana ruwan maniyyi da ƙara tsawon lokacin haihuwa.
An zabi namiji sharkasi mara-wuya mai mako 15 (homozygous rinjaye; Na Na) a matsayin masu ba da gudummawar maniyyi a cikin binciken.An yi kiwon tsuntsayen ne a gonar binciken kaji na tsangayar aikin gona, jami'ar Ashit, Ashit Governorate, Masar.An ajiye tsuntsaye a cikin cages guda ɗaya (30 x 40 x 40 cm), an ƙaddamar da shirin haske (16 hours na haske da 8 hours na duhu) da kuma ciyar da abincin da ke dauke da 160 g na furotin mai gina jiki, 2800 kcal na makamashi mai narkewa, 35 g na calcium kowanne.5 grams na samuwa phosphorus a kowace kilogiram na abinci.
Dangane da bayanai 36, 37, an tattara maniyyi daga maza ta hanyar tausa na ciki.An tattara jimlar samfuran maniyyi 45 daga maza 15 a cikin kwanaki 3.Maniyyi (n = 15 / day) nan da nan an diluted 1: 1 (v: v) tare da Belsville Poultry Semen Diluent, wanda ya ƙunshi potassium 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), potassium citrate monohydrate (0.064 g), potassium monophosphate (0.065 g), magnesium chloride (0.034 g) da H2O (100 ml), pH = 7, 5, osmolarity 3/333 mOsm.An fara bincika samfuran ruwan maniyyi a ƙarƙashin na'urar hangen nesa don tabbatar da ingancin maniyyi mai kyau (danshi) sannan a adana shi a cikin wanka na ruwa a 37 ° C har sai an yi amfani da shi a cikin rabin sa'a bayan tattarawa.
An kwatanta kinematics da rheology na spermatozoa ta amfani da tsarin na'urorin microfluidic.An ƙara ƙaddamar da samfurori na maniyyi zuwa 1: 40 a Beltsville Avian Semen Diluent, wanda aka ɗora a cikin na'urar microfluidic (duba ƙasa), kuma an ƙaddara sigogi na motsin rai ta amfani da tsarin nazarin maniyyi na Computerized (CASA) wanda aka tsara a baya don haɓaka halayen microfluidics.akan motsi na spermatozoa a cikin kafofin watsa labarai na ruwa (Sashen Injiniya 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 madaidaiciya (VSL, μm/s) da matsakaicin saurin yanayin (VAP, μm/s).An ɗauki bidiyon spermatozoa ta amfani da inverted Optika XDS-3 na'ura mai ba da hanya tsakanin hanyoyin sadarwa (tare da manufar 40x) da aka haɗa zuwa kyamarar Tucson ISH1000 a 30fps na 3 s.Yi amfani da software na CASA don nazarin aƙalla wurare uku da hanyoyin maniyyi 500 a kowane samfurin.An sarrafa bidiyon da aka yi rikodin ta amfani da CASA na gida.Ma'anar motsi a cikin plug-in CASA yana dogara ne akan saurin ninkaya na maniyyi idan aka kwatanta da yawan maniyyi, kuma baya haɗa da wasu sigogi kamar motsi gefe zuwa gefe, saboda an gano wannan ya fi dacewa da ruwa.An kwatanta motsin rheological a matsayin motsi na ƙwayoyin maniyyi a kan alkiblar ruwa.An raba spermatozoa tare da kaddarorin rheological ta yawan adadin spermatozoa mai motsi;spermatozoa wanda ke hutawa da kuma motsin spermatozoa mai motsi an cire su daga ƙidaya.
An samo duk sinadarai da aka yi amfani da su daga Elgomhoria Pharmaceuticals (Alkahira, Masar) sai dai in an lura da su.An kera na'urar kamar yadda El-sherry et al ya bayyana.40 tare da wasu gyare-gyare.Abubuwan da aka yi amfani da su don ƙirƙirar microchannels sun haɗa da faranti na gilashi (Howard Glass, Worcester, MA), SU-8-25 rashin ƙarfi (MicroChem, Newton, CA), barasa diacetone (Sigma Aldrich, Steinheim, Jamus), da polyacetone.-184, Dow Corning, Midland, Michigan).Ana ƙirƙira microchannels ta amfani da lithography mai laushi.Da farko, an buga abin rufe fuska mai tsabta tare da ƙirar microchannel da ake so akan firinta mai ƙarfi (Prismatic, Alkahira, Masar da Fasifik Arts and Design, Markham, ON).An yi masters ta hanyar amfani da faranti na gilashi a matsayin kayan aiki.An tsabtace faranti a cikin acetone, isopropanol da ruwa mai tsafta sannan kuma an rufe su da 20 µm Layer na SU8-25 ta suturar jujjuyawar (3000 rpm, 1 min).Su-8 yadudduka an bushe su a hankali (65°C, 2 min da 95°C, 10 min) kuma an fallasa su zuwa hasken UV na 50s.Gasa bayan fallasa a 65 ° C da 95 ° C don 1 min da 4 min don ƙetare yadudduka SU-8 da aka fallasa, sannan ci gaba a cikin barasa diacetone na 6.5 min.Gasa da waffles (200 ° C na minti 15) don ƙara ƙarfafa Layer SU-8.
An shirya PDMS ta hanyar haɗa monomer da hardener a cikin ma'aunin nauyi na 10: 1, sa'an nan kuma zazzage shi a cikin injin desiccator kuma a zuba a kan babban firam na SU-8.An warke PDMS a cikin tanda (120 ° C, 30 min), sannan an yanke tashoshi, an raba su da maigidan, kuma an huda su don ba da damar a haɗa bututu a mashigai da mashigar microchannel.A ƙarshe, ƙananan tashoshi na PDMS an haɗa su zuwa faifan microscope ta amfani da na'ura mai ɗaukar hoto na corona (Kayayyakin Fasaha na Electro-Technical, Chicago, IL) kamar yadda aka bayyana a wani wuri.Microchannel da aka yi amfani da shi a cikin wannan binciken yana auna 200 µm × 20 µm (W × H) kuma yana da tsayin 3.6 cm.
Ruwan ruwa wanda aka haifar da matsa lamba na hydrostatic a cikin microchannel yana samuwa ta hanyar kiyaye matakin ruwa a cikin tafki mai shiga sama da tsayin tsayin Δh39 a cikin tafki mai fita (Fig. 1).
inda f shine madaidaicin juzu'i, wanda aka bayyana a matsayin f = C / Re don kwararar laminar a cikin tashar rectangular, inda C ya kasance mai tsayi dangane da yanayin tashar tashar, L shine tsayin microchannel, Vav shine matsakaicin matsakaici a cikin microchannel, Dh shine diamita na hydraulic na tashar, g - haɓaka nauyi.Yin amfani da wannan ma'auni, ana iya ƙididdige matsakaicin saurin tashar ta amfani da ma'auni mai zuwa: