Daalụ maka ileta Nature.com.Ụdị ihe nchọgharị ị na-eji nwere nkwado CSS nwere oke.Maka ahụmịhe kachasị mma, anyị na-akwado ka ị jiri ihe nchọgharị emelitere (ma ọ bụ gbanyụọ ọnọdụ ndakọrịta na Internet Explorer).Ka ọ dị ugbu a, iji hụ na nkwado na-aga n'ihu, anyị ga-enye saịtị ahụ na-enweghị ụdị na Javascript.
Ọmụmụ nke nnụnụ na-adabere n'ikike ha nwere ịchekwa sperm zuru oke maka ogologo oge n'ime tubules nchekwa spam (SST).Usoro kpọmkwem nke spermatozoa na-abanye, na-ebi ma hapụ SST ka na-arụrịta ụka.Spam nke sharkasi hens gosipụtara ọchịchọ dị elu nke agglutination, na-akpụ ngwugwu filamentous mkpanaka nwere ọtụtụ mkpụrụ ndụ.N'ihi ihe isi ike nke ịhụ motility na omume nke spermatozoa na tube fallopian na-adịghị mma, anyị na-eji ngwaọrụ microfluidic nwere akụkụ cross microchannel yiri nke spermatozoa iji mụọ spermatozoa agglutination na motility.Ọmụmụ ihe a na-atụle ka sperm bundles na-etolite, ka ha si aga, na ọrụ ha nwere ike ime n'ịgbasa ebe obibi sperm na SST.Anyị nyochara ọsọ sperm na omume rheological mgbe mmiri na-emepụta n'ime ọwa microfluidic site na nrụgide hydrostatic (ọnụego mmiri = 33 µm/s).The spermatozoa na-achọkarị igwu mmiri megide ugbu a (ezi rheology) na ọsọ nke spermatozoon ngwugwu na-budata ebelata ma e jiri ya tụnyere otu spermatozoa.A hụla ngwugwu sperm ka ọ na-agagharị na gburugburu ma na-abawanye n'ogologo na ọkpụrụkpụ ka a na-ewebata otu spam. A hụrụ ngwugwu sperm ka ọ na-abịarute ma na-arapara n'akụkụ akụkụ nke ọwa microfluidic iji zere iji ọsọ mmiri na-asọ asọ> 33 µm/s. A hụrụ ngwugwu sperm ka ọ na-abịarute ma na-arapara n'akụkụ akụkụ nke ọwa microfluidic iji zere iji ọsọ mmiri na-asọ asọ> 33 µm/s. Было zamечено, что пучки сперматозоидов приближаются? ть сметания со скоростью потока жидкости> 33 мкм / с. A hụla ngwugwu sperm ka ọ na-abịarute ma na-arapara n'akụkụ mgbidi nke ọwa microfluidic iji zere ikpochapụ na ọnụego mmiri mmiri> 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.33 µm/s 扫过. Было zamечено, что пучки сперматозоидов приближаются ь сметания потоком жидкости со скоростью > 33 мкм/с. A hụla ngwugwu sperm ka ọ na-abịarute ma na-arapara n'akụkụ mgbidi nke ọwa microfluidic ka ọ ghara ikpochapụ site na mmiri mmiri na> 33 µm/s.Nyochaa na nnyefe eletrọnịkị na-egosi na ejiri nnukwu ihe na-akwado ùkwù sperm.Ihe data enwetara na-egosi mmegharị pụrụ iche nke Sharkazi chicken spermatozoa, yana ikike nke spermatozoa na agglutinate na ịmepụta ngwugwu mkpanaka, nke na-eme ka nghọta dịkwuo mma nke nchekwa ogologo oge nke spermatozoa na SMT.
Iji nweta fatịlaịza n'ime mmadụ na ọtụtụ anụmanụ, sperm na àkwá ga-erurịrị ebe a na-amụba n'oge kwesịrị ekwesị.Ya mere, mating ga-eme tupu ma ọ bụ n'oge ovulation.N'aka nke ọzọ, ụfọdụ anụ mammals, dị ka nkịta, yana ụdị ndị na-abụghị anụ mammali, dị ka ụmụ ahụhụ, azụ, anụ ufe, na nnụnụ, na-echekwa spam n'ime akụkụ ahụ ha ọmụmụ ruo ogologo oge ruo mgbe akwa ha dị njikere maka fatịlaịza (asynchronous Fertilization 1).Nnụnụ na-enwe ike ịnọgide na-enwe ike nke spermatozoa ike nke fatịlaịza àkwá maka 2-10 weeks2.
Nke a bụ ihe pụrụ iche nke na-eme ka nnụnụ dị iche na anụmanụ ndị ọzọ, n'ihi na ọ na-enye ohere dị elu nke fatịlaịza mgbe otu izu gasịrị ruo ọtụtụ izu na-enweghị n'otu oge mating na ovulation.Isi ihe nchekwa sperm, nke a na-akpọ tubule sperm storage tubule (SST), dị na mpịakọta mucosal dị n'ime na nkwụsị uterovaginal.Ka ọ dị ugbu a, usoro nke spam na-esi abanye, biri, na ọpụpụ n'ụlọ akụ spam na-aghọtachaghị nke ọma.Dabere na ọmụmụ ndị gara aga, e tinyela ọtụtụ echiche n'ihu, ma ọ dịghị nke ọ bụla n'ime ha ka akwadoro.
Forman4 chere na spermatozoa na-edobe ebe obibi ha na oghere SST site na mmegharị oscillatory na-aga n'ihu megide ntụzịaka nke mmiri na-asọ site na ọwa protein dị na SST epithelial cell (rheology).ATP na-agwụcha n'ihi ọrụ flagellar na-adịgide adịgide nke achọrọ iji mee ka spam na-adị na SST lumen na motility na-emecha daa ruo mgbe spam na-esi na mmiri na-asọpụta na-esi na mmiri pụta wee malite njem ọhụrụ na-agbada na tube nke fallopian na-arịgoro iji fatịlaịza spam.Akwa (Forman4).Ihe nlereanya a nke nchekwa spam na-akwado site na nchọpụta nke immunocytochemistry nke aquaporins 2, 3 na 9 dị na sel SST epithelial.Ka ọ dị ugbu a, ọmụmụ banyere rheology semen ọkụkọ na ọrụ ya na nchekwa SST, nhọrọ sperm vaginal, na asọmpi spam adịghị.N'ime ọkụkọ, spam na-abanye n'ime ikpu mgbe mmekọ nwoke na nwanyị gachara, mana ihe karịrị 80% nke spermatozoa na-esi na ikpu pụta obere oge ka ha nwesịrị.Nke a na-egosi na ikpu bụ ebe bụ isi maka ịhọrọ sperm na nnụnụ.Na mgbakwunye, a kọwapụtara na ihe na-erughị 1% nke spermatozoa fatịlaịza na ikpu na-ejedebe na SSTs2.Na artificial insemination nke chicks na ikpu, ọnụ ọgụgụ nke spermatozoa eru SST na-emekarị na-abawanye 24 awa mgbe insemination.Ka ọ dị ugbu a, usoro a na-ahọrọ sperm n'oge usoro a amabeghị, na motility spam nwere ike ịrụ ọrụ dị mkpa n'ịkwalite sperm SST.N'ihi nnukwu mgbidi na opaque nke tubes fallopian, ọ na-esiri ike nyochaa ozugbo spam na-agagharị na tubes fallopian nke nnụnụ.Ya mere, anyị enweghị isi ihe ọmụma banyere otú spermatozoa na-aga na SST mgbe fatịlaịza.
Achọpụtala Rheology n'oge na-adịbeghị anya dị ka ihe dị mkpa na-achịkwa mbufe spam na amụ nke anụ mammalian.Dabere na ikike spermatozoa motile ịkwaga n'oge na-adịbeghị anya, Zaferani et al8 jiri sistemu corra microfluidic kewapụ spermatozoa motile n'ụdị semen edere.Ụdị nhazi nke mmanụ a dị mkpa maka ọgwụgwọ amụghị nwa na nyocha ụlọ ọgwụ, a na-ahọrọ ya karịa usoro omenala bụ oge na ọrụ siri ike ma nwee ike imebi sperm morphology na iguzosi ike n'ezi ihe.Otú ọ dị, ruo taa, ọ dịghị nnyocha e mere na mmetụta nke secretions si genital akụkụ nke ọkụkọ na spam motility.
N'agbanyeghị usoro nke na-edobe spam na-echekwa na SST, ọtụtụ ndị nchọpụta achọpụtala na spermatozoa agglutinate bi na isi na isi na SST nke ọkụkọ 9, 10, quails 2, na turkeys 11 iji mepụta sperm bundles agglutinated.Ndị na-ede akwụkwọ na-atụ aro na e nwere njikọ n'etiti agglutination a na nchekwa ogologo oge nke spermatozoa na SST.
Tingari na Lake12 kọrọ njikọ siri ike n'etiti spermatozoa na gland nke ọkụkọ na-anata spam ma jụọ ma avian spermatozoa agglutinate n'otu ụzọ ahụ dị ka mammalian spermatozoa.Ha kwenyere na njikọ miri emi dị n'etiti sperm na vas deferens nwere ike ịbụ n'ihi nrụgide nke ọnụnọ nke ọnụ ọgụgụ dị ukwuu nke sperm na obere oghere.
Mgbe a na-enyocha omume spermatozoa na slide iko iko ọhụrụ, enwere ike ịhụ ihe ịrịba ama na-adịru nwa oge nke agglutination, ọkachasị n'akụkụ ọnụ mmiri nke semen.Otú ọ dị, agglutination na-enwekarị nchekasị site n'omume ntụgharị nke jikọtara ya na mmegharị ahụ na-aga n'ihu, nke na-akọwa ọdịdị na-adịte aka nke ihe a.Ndị nchọpụta ahụ chọpụtakwara na mgbe agbakwunyere ihe na-esi ísì ụtọ na semen, nchịkọta cell "string-like" elongated pụtara.
E mere mgbalị mbụ iji ṅomie spermatozoon site n'iwepụ waya dị gịrịgịrị n'ime ndopu nghọta, nke butere vesicle elongated sperm dị ka vesicle na-apụta site na ndo nke semen.Na spermatozoa ozugbo kwụ n'ahịrị n'usoro yiri nke ahụ n'ime vesicle, ma akụkụ ahụ dum kwụsịrị ngwa ngwa n'ihi njedebe 3D.Ya mere, iji mụọ spermatozoa agglutination, ọ dị mkpa ịhụ motility na omume nke spermatozoa ozugbo na ebe nchekwa sperm tubules, nke siri ike nweta.Ya mere, ọ dị mkpa ịmepụta ngwá ọrụ na-eṅomi spermatozoa iji kwado ọmụmụ ihe gbasara motility sperm na agglutination omume.Brillard et al13 kwuru na nkezi ogologo tubules nchekwa spam na ụmụ ọkụkọ toro eto bụ 400-600 µm, mana ụfọdụ SST nwere ike ịdị ogologo 2000 µm.Mero na Ogasawara14 kewara glands semiferous ka ọ bụrụ tubules nchekwa spam na-agbasawanye na nke na-abụghị nnukwu, ha abụọ bụ otu n'ogologo (~ 500 µm) na obosara olu (~ 38 µm), mana obosara lumen nke tubules bụ 56.6 na 56.6 µm.., otu 11.2 μm, n'otu n'otu.N'ime ọmụmụ ihe ugbu a, anyị na-eji ngwaọrụ microfluidic nwere ọwa nke 200 µm × 20 µm (W × H), nke akụkụ obe dị ntakịrị nso na nke SST.Na mgbakwunye, anyị nyochara motility sperm na agglutination omume na mmiri na-asọpụta, nke kwekọrọ n'echiche Foreman na mmiri mmiri nke SST epithelial cell na-emepụta na-edobe spam na lumen na ntụgharị ihu (rheological).
Ebumnuche nke ọmụmụ a bụ imeri nsogbu nke ịhụ spermatozoa motility na tube fallopian na iji zere ihe isi ike nke ịmụ gbasara rheology na omume nke spermatozoa na gburugburu ebe dị ike.A na-eji ngwaọrụ microfluidic nke na-emepụta nrụgide hydrostatic iji mee ka motility spam dị n'ime akụkụ ahụ nke ọkụkọ.
Mgbe etinyere dobe nke sperm sample (1:40) n'ime ngwaọrụ microchannel, a ga-achọpụta ụdị motility spam abụọ (sperm dịpụrụ adịpụ na bound sperm).Tụkwasị na nke ahụ, spermatozoa na-achọ igwu mmiri megide ihe dị ugbu a (ezigbo rheology; vidiyo 1, 2). Ọ bụ ezie na ngwugwu sperm nwere obere ọsọ karịa nke spam naanị (p <0.001), ha mụbara pasentị nke spam na-egosipụta ezigbo rheotaxis (p <0.001; Tebụl 2). Ọ bụ ezie na ngwugwu sperm nwere obere ọsọ karịa nke spam naanị (p <0.001), ha mụbara pasentị nke spam na-egosipụta ezigbo rheotaxis (p <0.001; Tebụl 2). Хотя пучки сперматозоидов имели более низкую скорость. ерматозоидов, демонстрирующих положительный реотаксис (p <0,001; таблица 2). Ọ bụ ezie na ngwugwu spermatozoa nwere obere ọsọ karịa nke otu spermatozoa (p <0.001), ha mụbara pasent nke spermatozoa na-egosi rheotaxis dị mma (p <0.001; Table 2).精子束 的 的 速度 孤独 精子 精子 孤独 精子 精子 0.001), 但 它们 显示 阳性 的 精子 精子 精子 精子 精子 精子 0.001; 表 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. дов с положительной реологией (p <0,001; таблица 2). Ọ bụ ezie na ọsọ nke ngwugwu sperm dị ala karịa nke otu spermatozoa (p <0.001), ha mụbara pasent nke spermatozoa na rheology dị mma (p <0.001; Table 2).A na-eme atụmatụ rheology dị mma maka otu spermatozoa na tufts na ihe dịka 53% na 85%, n'otu n'otu.
Achọpụtala na spermatozoa nke ọkụkọ sharkasi ozugbo ejaculation gasịrị na-etolite ngwugwu linear, nke nwere ọtụtụ mmadụ.Tufts ndị a na-abawanye ogologo na ọkpụrụkpụ ka oge na-aga ma nwee ike ịnọ na vitro ruo ọtụtụ awa tupu ịgbasa (video 3).Ihe ngwugwu filamentous ndị a na-eme ka echidna spermatozoa nke na-etolite na njedebe nke epididymis.A chọpụtala na mma mmiri Sharkashi hen nwere nnukwu ọchịchọ nke agglutinate ma mepụta ngwugwu reticulate n'ihe na-erughị otu nkeji mgbe nchịkọta.Osisi ndị a dị ike ma nwee ike ịrapara na mgbidi ọ bụla dị nso ma ọ bụ ihe kwụ ọtọ.Ọ bụ ezie na ngwugwu sperm na-ebelata ọsọ nke mkpụrụ ndụ sperm, o doro anya na macroscopically ha na-abawanye linearity ha.Ogologo ùkwù ahụ dịgasị iche dabere na ọnụ ọgụgụ spam anakọtara na ngwugwu.Akụkụ abụọ nke ngwugwu ahụ dịpụrụ adịpụ: akụkụ mbụ, gụnyere isi free nke spam agglutinated, na akụkụ njedebe, gụnyere ọdụ na njedebe njedebe nke spam.N'iji igwefoto dị elu (950fps), a na-ahụ isi n'efu nke agglutinated spermatozoa na akụkụ mbụ nke ngwugwu ahụ, na-ahụ maka mmegharị nke ngwugwu ahụ n'ihi mmegharị oscillatory ha, na-adọkpụrụ ndị fọdụrụ n'ime ngwugwu ahụ na mmegharị helical (Video 4).Otú ọ dị, n'ime ogologo tuft, a chọpụtala na ụfọdụ isi spam na-akwụghị ụgwọ na-arapara n'ahụ ahụ na akụkụ njedebe nke tuft ahụ na-eme ihe dị ka vanes na-enyere aka ịkwasa tuft.
N'ịbụ ndị na-eji nwayọọ nwayọọ na-agbapụta mmiri, sperm bundles na-aga n'ihu na ibe ha, Otú ọ dị, ha na-amalite ịgbakọ ma rapara n'ihe niile dị ka, ka mmiri na-agba ugbu a ghara ịsacha ya ka ọsọ ọsọ na-abawanye.Mkpokọta ndị ahụ na-etolite mgbe ntakịrị mkpụrụ ndụ spam na-abịaru ibe ha nso, ha na-amalite ịkwagharị na synchrony wee kechie ibe ha, wee rapara na ihe na-egbuke egbuke.Ihe onyonyo nke 1 na nke 2 na-egosi ka spam na-esi abịaru ibe ha nso, na-eme njikọ ka ọdụ na-ekekọta ibe ha.
Ndị nchọpụta ahụ tinyere nrụgide hydrostatic iji mepụta mmiri na-asọ na microchannel iji mụọ rheology sperm.Eji microchannel nwere nha 200µm × 20 µm (W × H) yana ogologo 3.6 µm.Jiri microchannel n'etiti arịa nwere sirinji dabara na nsọtụ.Ejiri agba agba nri mee ka ọwa ndị ahụ pụta ìhè.
Kee eriri njikọ njikọ na ngwa na mgbidi.Ejiri microscope dị iche iche were were were were se vidiyo a.N'ihe onyonyo ọ bụla, a na-ewepụta ihe onyonyo dị iche na nkewa.(A) Njikọ dị n'etiti iyi abụọ na-eguzogide mmiri ahụ n'ihi mmegharị helical (akụ uhie).(B) Njikọ dị n'etiti ngwugwu tube na mgbidi ọwa (akụ uhie), n'otu oge ahụ ha na-ejikọta na ngwugwu abụọ ọzọ (akụ na-acha odo odo).(C) Ngwunye sperm na ọwa microfluidic na-amalite ijikọ na ibe ya (akụ uhie), na-akpụ nchikota nke sperm bundles.(D) Ịmepụta netwọk nke ngwugwu sperm.
Mgbe etinyere dobe nke sperm diluted n'ime ngwaọrụ microfluidic ma mepụta mmiri na-asọpụta, a na-ahụ oghere spam na-aga n'ihu na-aga n'ihu.Ngwunye ndị ahụ dabara adaba na mgbidi nke microchannels, na isi ndị nweere onwe ha na akụkụ mbụ nke ngwugwu dabara adaba na ha (video 5).Ha na-araparakwa n'ihe ọ bụla guzoro n'ụzọ ha, dị ka irighiri ihe, ka ihe ndị dị ugbu a na-ekpochapụ.Ka oge na-aga, ndị a tufts na-aghọ ogologo filaments trapping ndị ọzọ otu spermatozoa na mkpumkpu tufts (Video 6).Ka mmiri na-amalite iji nwayọọ nwayọọ, ogologo ahịrị sperm na-amalite ịmalite netwọk nke ahịrị sperm (Video 7; Figure 2).
N'ọsọ ọsọ dị elu (V> 33 µm/s), a na-abawanye mmegharị nke eriri dị ka mbọ iji jide ọtụtụ sperm nke na-emepụta ngwugwu ka mma iguzogide ike na-ebugharị. N'ọsọ ọsọ dị elu (V> 33 µm/s), a na-abawanye mmegharị nke eriri dị ka mbọ iji jide ọtụtụ sperm nke na-emepụta ngwugwu ka mma iguzogide ike na-ebugharị. При высокой скорости потока (V > 33 мкм/с) спиралевидные тдельных сперматозоидов, образующих пучки, которые лучше противостоят дрейфующей силе потока. N'ọnụego dị elu (V> 33 µm / s), mmegharị helical nke strands na-abawanye ka ha na-agbalị ijide ọtụtụ spermatozoa n'otu n'otu na-akpụ ùkwù ndị na-enwe ike iguzogide ike na-ebugharị nke mmiri ahụ.在高流过流动的漂移力。在 高 流速 (v> 33 µm/s) 时 ， 的 螺旋 运动 增加 ，抗 的 漂移力………………………….. При высоких скоростях потока (V > 33 мкм/с) матозоидов, образующих пучки, чтобы лучше сопротивляться силам дрейфа потока. Na ọnụ ọgụgụ dị elu (V> 33 µm / s), mmegharị ahụ nke filaments na-abawanye na mbọ iji weghara ọtụtụ spermatozoa nke na-emepụta ngwugwu ka mma iguzogide ike na-ebugharị nke mmiri ahụ.Ha gbalịkwara itinye microchannel n'akụkụ mgbidi.
Achọpụtara ngwugwu sperm dị ka ụyọkọ isi spam na ọdụ curling site na iji microscopy ìhè (LM).Achọpụtakwara ngwugwu sperm nwere nchịkọta dị iche iche dị ka isi gbagọrọ agbagọ na mkpokọta flagellar, ọtụtụ ọdụ spam jikọtara ọnụ, isi sperm nke etinyere na ọdụ, na isi spam nwere nuclei gbagoro agbagọ dị ka ọtụtụ ihe jikọrọ ọnụ.nnyefe eletrọnịkị microscope (TEM).Nnyocha eletrọnịkị microscopy (SEM) gosiri na ùkwù sperm ahụ bụ ngwakọta nke isi sperm n'ọbọ, na mkpokọta sperm na-egosi njikọ agbakwunyere nke ọdụ ọbọp.
A na-amụ morphology na ultrastructure nke spermatozoa, nhazi nke spermatozoa bundles site na iji microscopy ìhè (ọkara ngalaba), nyocha microscopy eletrọnịkị (SEM) na microscopy eletrọnịkị (TEM), sperm smears na-acha acridine oroma wee nyochaa site na iji epifluorescence microscopy.
Smear smear staining na acridine orange (Fig 3B) gosiri na isi spam na-ejikọta ọnụ ma kpuchie ya na ihe nzuzo, nke mere ka e nwee nnukwu tufts (Fig 3D).Ngwunye sperm ahụ nwere nchịkọta sperm na netwọk ọdụ ọdụ (Fig 4A-C).Ihe mejupụtara ùkwù spam bụ ọdụ nke ọtụtụ spermatozoa rapaara ọnụ (Fig 4D).Ihe nzuzo (Fig 4E,F) kpuchiri isi nke ngwugwu spermatozoa.
Nhazi nke spermatozoa ngwugwu Iji frasi iche microscopy na spam smears nwere acridine oroma, gosiri na isi nke spermatozoa na-arapara ọnụ.(A) Nhazi sperm tuft n'oge na-amalite site na spam (ọcha okirikiri) na spam atọ (okirikiri odo), na gburugburu na-amalite na ọdụ na-ejedebe na isi.(B) Foto foto nke sperm smear nwere ntụ oroma acridine na-egosi isi spam na-ejikọta ya (akụ).Mwepu na-ekpuchi isi (s).Magnification × 1000. (C) Mmepe nke nnukwu ọkụ na-ebufe site na ntinye na ọwa microfluidic (iji igwefoto dị elu na 950 fps).(D) Micrograph nke sperm smear nwere ntụ oroma acridine na-egosi nnukwu tufts (akụ).Nkwalite: ×200.
Na-enyocha micrograph eletrọn nke oghere sperm na smear spam nwere ntụ oroma acridine.(A, B, D, E) bụ ihe na-enyocha agba dijitalụ nke micrographs nke spermatozoa, na C na F bụ micrographs nke acridine oroma nwere ntụpọ spam na-egosi ntinye nke otutu spermatozoa na-ekpuchi webụ caudal.(AC) A na-egosi mkpokọta spam dị ka netwọk ọdụ ọdụ (akụ).(D) Mgbakwunye nke ọtụtụ spermatozoa (nke nwere ihe nrapado, akara pink, akụ) na-ekpuchi ọdụ.(E na F) Mkpokọta isi sperm (pointers) ejiri ihe nrapado kpuchie (atụ).Akpụkpọ anụ spermatozoa nwere ọtụtụ ihe dị ka vortex (F).(C) × 400 na (F) × 200 mmụba.
N'iji microscopy eletrọn nnyefe, anyị chọpụtara na ngwugwu sperm nwere ọdụ ọdụ (Fig 6A, C), isi ndị na-ejikọta na ọdụ (Fig 6B), ma ọ bụ isi na-ejikọta ọdụ (Fig 6D).Ndị isi nke spermatozoa na ngwugwu na-agbagọ, na-egosi na ngalaba abụọ mpaghara nuklia (Fig 6D).Na ngwugwu mbepụ, spermatozoa nwere isi gbagọrọ agbagọ nke nwere mpaghara nuklia abụọ na mpaghara ọtụtụ flagellar (Fig 5A).
Igwe eletrọn ụcha dijitalụ na-egosi ọdụ njikọ dị na ngwugwu spam na ihe agglutinating na-ejikọta isi spam.(A) Ejikọtara ọdụ nke ọnụ ọgụgụ buru ibu nke spermatozoa.Rịba ama ka ọdụ ahụ si dị na ntule eserese (akụ) na n'ọdịdị ala (akụ).(B) Isi (akụ) nke spam na-ejikọta na ọdụ (akụ).(C) A na-ejikọta ọtụtụ ọdụ ọdụ spam (akụ).(D) Agglutination ihe (AS, blue) jikọtara isi spam anọ (acha odo odo).
A na-eji nyocha eletrọn microscopy chọpụta isi spam na ngwugwu sperm kpuchiri ihe nzuzo ma ọ bụ akpụkpọ anụ (Njirimara 6B), nke na-egosi na sperm bundles bụ ihe extracellular kwụchiri.A na-etinye ihe agglutinated na isi spam (jellyfish isi-dị ka mgbakọ; Fig. 5B) ma gbasaa n'ebe dị anya, na-enye ọdịdị na-acha odo odo na-egbuke egbuke n'okpuru microscopy fluorescence mgbe ejiri ya na acridine oroma (Fig 6C).A na-ahụ ihe a nke ọma n'okpuru microscope na-enyocha ma na-ewere ya dị ka ihe nkedo.Akụkụ dị obere (Fig 5C) na sperm smears nwere ntụ oroma acridine gosiri ngwugwu spam nwere isi juru n'ọnụ na ọdụ mpịaji (Fig 5D).
Foto dị iche iche na-egosi nchịkọta isi spam na ọdụ apịaji site na iji ụzọ dị iche iche.(A) Cross-section dijitalụ agba nnyefe eletrọn micrograph nke ngwugwu spam na-egosi isi sperm gbakwụnyere nke nwere akụkụ abụọ nwere akụkụ (acha anụnụ anụnụ) na ọtụtụ akụkụ flagellar (green).(B) Micrograph eletrọn na-enyocha ụcha dijitalụ na-egosi ụyọkọ isi spam (akụ) nke yiri ka ekpuchiri ya.(C) Nkeji dị obere nke na-egosi isi sperm ekpokọtara ọnụ (akụ) na ọdụ gbachiri agbakọ (akụ).(D) Micrograph nke sperm smear nke nwere acridine oroma na-egosi mkpokọta isi spam (akụ) na ọdụ na-agbachitere (akụ).Rịba ama na ihe nnyapade (S) na-ekpuchi isi nke spermatozoon.(D) × 1000 mmụba.
Iji nnyefe eletrọnịkị microscopy (Fig 7A), a chọpụtakwara na isi spam na-agbagọ na nuclei nwere ọdịdị okirikiri, dị ka sperm smears nwere ntụpọ acridine na-enyocha ya na iji fluorescence microscopy (Fig 7B).
(A) Digital agba nnyefe eletrọn micrograph na (B) Acridine oroma ntụpọ spam smear na-egosi chikọtara isi na mgbakwunye nke spam isi na ọdụ (akụ).(B) × 1000 mmụba.
Ihe nchoputa na-adọrọ mmasị bụ na sperm Sharkazi na-achịkọta ọnụ iji mepụta ngwugwu filamentous mkpanaka.Njirimara nke ngwugwu ndị a na-enye anyị ohere ịghọta ọrụ ha nwere ike ime na absorption na nchekwa nke spermatozoa na SST.
Mgbe nwoke na nwanyị gachara, spam na-abanye n'ime ikpu ma na-eme usoro nhọrọ siri ike, nke na-ebute naanị ọnụ ọgụgụ spam na-abanye na SST15,16.Ka ọ dị ugbu a, usoro nke spam na-esi abanye ma pụta na SST amabeghị.N'ime anụ ọkụkọ, a na-echekwa spermatozoa na SST maka ogologo oge nke izu 2 ruo 10, dabere na ụdị6.Esemokwu na-anọgide banyere ọnọdụ nke semen n'oge nchekwa na SST.Ha na-emegharị ma ọ bụ na-ezu ike?N'ikwu ya n'ụzọ ọzọ, kedu ka mkpụrụ ndụ sperm si edebe ọnọdụ ha na SST ogologo oge?
Forman4 tụrụ aro na enwere ike ịkọwa ebe obibi SST na nchụpụ n'ihe gbasara motility spam.Ndị na-ede akwụkwọ na-eche na spam na-edobe ọnọdụ ha site na igwu mmiri megide mmiri mmiri nke SST epithelium na-emepụta na spam na-esi na SST pụta mgbe ọsọ ha dara n'okpuru ebe ha na-amalite ịlaghachi azụ n'ihi enweghị ike.Zaniboni5 kwadoro ọnụnọ nke aquaporins 2, 3, na 9 na akụkụ apical nke SST epithelial cell, nke nwere ike ịkwado ụdị nchekwa spam nke Foreman n'ezoghị ọnụ.N'ime ọmụmụ ihe ugbu a, anyị chọpụtara na ihe fọrọ nke nta ka ọ bụrụ ọkara nke Sharkashi's spermatozoa na-egosi rheology dị mma na mmiri na-asọpụta, na nchịkọta spermatozoa na-abawanye ọnụ ọgụgụ nke spermatozoa na-egosi rheology dị mma, ọ bụ ezie na agglutination na-ebelata ha.Aghọtachaghị etu mkpụrụ ndụ sperm si agbago tube fallopian nke nnụnụ na saịtị spam nwoke na nwanyị.N'ime ụmụ anụmanụ, mmiri follicular chemo na-adọta spermatozoa.Agbanyeghị, ekwenyere na chemoattractants na-eduzi spermatozoa ka ọ bịaruo ebe dị anya7.Ya mere, usoro ndị ọzọ na-ahụ maka ibuga spam.Ikike nke spam na-atụgharị ma na-asọba n'ime mmiri nke tube nke fallopian nke ewepụtara ka a na-akọ na ọ bụ isi ihe na-eme ka sperm dị na ụmụ oke.Parker 17 tụrụ aro ka spermatozoa gafere oviducts site n'igwu mmiri megide oke mmiri nke anụ ufe na anụ ufe.Ọ bụ ezie na e gosipụtabeghị ya na nnwale na nnụnụ, Adolphi18 bụ onye mbụ chọpụtara na spam na-enye nsonaazụ dị mma mgbe ejiri mpempe akwụkwọ nzacha mepụta obere mmiri mmiri dị n'etiti ihe mkpuchi na slide.Rheology.Hino na Yanagimachi [19] tinyere mgbagwoju ovary-tubal-uterine òké n'ime mgbanaka perfusion wee gbanye 1 µl nke ink n'ime isthmus iji hụ ka mmiri na-asọ na tubes fallopian.Ha hụrụ mmegharị ahụ na-arụsi ọrụ ike nke mkpụkọ na izu ike na tube nke fallopian, bụ nke bọọlụ ink niile na-aga n'ihu na-aga n'ihu na ampulla nke tube fallopian.Ndị ode akwụkwọ na-ekwusi ike na ọ dị mkpa ka mmiri mmiri na-agbapụta site na ala ruo n'ime akpa fallopian nke elu maka ebuli sperm na fatịlaịza.Brillard20 kọrọ na na ọkụkọ na turkeys, spermatozoa na-akwaga site na-arụsi ọrụ ike ije si n'ọnụ ụzọ ikpu, ebe a na-echekwara ha, na utero-ikpu njikọ, ebe a na-echekwara ha.Otú ọ dị, a dịghị achọ mmegharị a n'etiti njikọ uterovaginal na infundibulum n'ihi na a na-ebufe spermatozoa site na nchụpụ na-agafe agafe.N'ịmara ndụmọdụ ndị a gara aga na nsonaazụ ndị a na-enweta na ọmụmụ ihe ugbu a, enwere ike iche na ike nke spermatozoa ịkwaga elu (rheology) bụ otu n'ime ihe ndị a na-eme ka usoro nhọrọ dabere na ya.Nke a na-ekpebi ngafe nke spermatozoa site na ikpu na ntinye ha na CCT maka nchekwa.Dị ka Forman4 tụrụ aro, nke a nwekwara ike ime ka usoro spam na-abanye na SST na ebe obibi ya ruo oge ụfọdụ wee pụọ mgbe ọsọ ha malitere iji nwayọọ.
N'aka nke ọzọ, Matsuzaki na Sasanami 21 tụrụ aro na avian spermatozoa na-enwe mgbanwe motility site na dormancy na motility na traktị ọmụmụ nwoke na nwanyị.Mgbochi nke sperm motility bi na SST ka akwadoro iji kọwaa ogologo oge nchekwa nke spam na mgbe ahụ rejuvenation mgbe ọ hapụsịrị SST.N'okpuru ọnọdụ hypoxic, Matsuzaki et al.1 kọrọ nnukwu mmepụta na mwepụta nke lactate na SST, nke nwere ike iduga na mgbochi nke motility sperm bi.N'okwu a, mkpa sperm rheology na-egosipụta na nhọrọ na absorption nke spermatozoa, ọ bụghị na nchekwa ha.
A na-ewere ụkpụrụ agglutination sperm dị ka nkọwa ziri ezi maka ogologo oge nchekwa spam na SST, n'ihi na nke a bụ ihe a na-ahụkarị nke njide spam na anụ ọkụkọ2,22,23.Bakst et al.2 chọpụtara na ọtụtụ spermatozoa na-arapara n'ahụ ibe ha, na-emepụta mkpokọta fascicular, na otu spermatozoa adịghị adịkarị na quail CCM.N'aka nke ọzọ, Wen et al.24 hụrụ spermatozoa gbasasịa karịa na obere spermatozoa tufts na SST lumen na ọkụkọ.Dabere na nlebanya ndị a, enwere ike iche na mmasị maka spam agglutination dị iche n'etiti nnụnụ na n'etiti spermatozoa n'otu ejaculate.Na mgbakwunye, Van Krey et al.9 na-atụ aro na enweghị nkewa nke agglutinated spermatozoa bụ maka ntinye nwayọọ nwayọọ nke spermatozoa n'ime lumen nke tube fallopian.Dị ka echiche a si kwuo, ekwesịrị ịchụpụ spermatozoa nwere ikike agglutination dị ala na SST mbụ.N'okwu a, ikike nke spermatozoa na agglutinate nwere ike ịbụ ihe na-emetụta nsonaazụ asọmpi spam na nnụnụ ruru unyi.Na mgbakwunye, ogologo oge sperm agglutinated na-ekewa, ogologo oge ọmụmụ ka a na-edobe.
Ọ bụ ezie na spermatozoa aggregation na aggregation n'ime ngwugwu ka ahụrụ n'ọtụtụ ọmụmụ2,22,24, a kọwabeghị ha n'ụzọ zuru ezu n'ihi mgbagwoju anya nke kinematic ha na-ahụ n'ime SST.Emeela ọtụtụ mbọ iji mụọ sperm agglutination in vitro.A hụrụ mkpokọta sara mbara mana na-adịte aka mgbe e wepụrụ waya dị mkpa na dobe mkpụrụ na-akụ.Nke a na-eduga n'eziokwu ahụ bụ na elongated afụ na-esi na dobe ahụ pụta, na-eṅomi gland seminal.N'ihi oke 3D na oge ihicha mmiri dị mkpụmkpụ, ngọngọ ahụ niile dabara ngwa ngwa9.N'ime ọmụmụ ihe ugbu a, na-eji ọkụkọ Sharkashi na ibe microfluidic, anyị nwere ike ịkọwa otú tufts ndị a si etolite na otu ha si aga.Mpempe sperm na-etolite ozugbo nchịkọta mkpụrụ ndụ ma chọpụta na ọ na-agagharị na gburugburu, na-egosi rheology dị mma mgbe ọ dị na mmiri.Ọzọkwa, mgbe elele anya macroscopically, a hụla ngwungwu sperm na-abawanye linearity nke motility ma e jiri ya tụnyere spermatozoa dịpụrụ adịpụ.Nke a na-egosi na sperm agglutination nwere ike ime tupu ntinye SST na mmepụta spam na-ejedebe na obere mpaghara n'ihi nrụgide dị ka a tụrụ aro na mbụ (Tingari na Lake12).N'oge a na-emepụta tuft, spermatozoa na-egwu mmiri na synchrony ruo mgbe ha na-eme njikọ, wee kechie ọdụ ha gburugburu ibe ha na isi nke spermatozoon na-anọgide na-enwere onwe ya, ma ọdụ na akụkụ dị anya nke spermatozoon na-ejikọta ya na ihe na-egbuke egbuke.Ya mere, isi n'efu nke ligament na-ahụ maka mmegharị ahụ, na-adọkpụrụ ndị ọzọ nke ligament.Nyochaa microscopy eletrọn nke sperm bundles gosiri isi spam na-ejikọta ya na ọtụtụ ihe na-atọ ụtọ, na-atụ aro na isi spam na-ejikọta ya na ngwugwu izu ike, nke nwere ike ime mgbe ha rutere ebe nchekwa (SST).
Mgbe sperm smear na-acha acridine oroma, a pụrụ ịhụ ihe nrapado extracellular gburugburu mkpụrụ ndụ sperm n'okpuru microscope fluorescent.Ihe a na-eme ka ngwugwu spam na-arapara ma rapara n'elu elu ma ọ bụ ihe ọ bụla gbara ya gburugburu ka ha ghara iji mmiri na-agba gburugburu.Ya mere, ihe anyị chọpụtara na-egosi ọrụ nke spermatozoa adhesion n'ụdị nke mobile bundles.Ikike ha igwu mmiri megide ihe dị ugbu a na ịrapara n'akụkụ dị nso na-enye ohere ka spam na-anọ ogologo oge na SST.
Rothschild25 ji igwefoto hemocytometry amụ banyere nkesa nke semen bovine na-ese n'elu mmiri na nkwusioru nke nkwusioru, na-ewere fotomicrograph site na igwefoto nwere ma axis optical vetikal na kwụ ọtọ nke microscope.Nsonaazụ gosiri na spermatozoa na-adọta n'elu ụlọ ahụ.Ndị na-ede akwụkwọ na-atụ aro na enwere ike inwe mmekọrịta hydrodynamic n'etiti spam na elu.N'iburu nke a n'uche, yana ikike nke Sharkashi chick semen na-emepụta tufts na-egbuke egbuke, ọ nwere ike ịbawanye ohere na mmanụ aṅụ ga-arapara na mgbidi SST ma chekwaa ya ogologo oge.
Bccetti na Afzeliu26 kọrọ na a chọrọ spam glycocalyx maka njirimara gamete na agglutination.Forman10 chọpụtara na hydrolysis nke α-glycosidic bonds na glycoprotein-glycolipid mkpuchi site n'ịgwọ semen avian na neuraminidase mere ka mbelata ọmụmụ na-enweghị emetụta mmụgharị spam.Ndị na-ede akwụkwọ na-atụ aro na mmetụta nke neuraminidase na glycocalyx na-emebi sperm sequestration na utero-vaginal junction, si otú ibelata ọmụmụ.Nleba anya ha enweghị ike ileghara ohere na ọgwụgwọ neuraminidase nwere ike ibelata spam na nnabata oocyte.Forman na Engel10 chọpụtara na ọmụmụ na-ebelata mgbe a na-ekesa ọkụkọ intravaginal site na semen ejiri neuraminidase gwọọ ya.Otú ọ dị, IVF nwere neuraminidase na-agwọ sperm emetụtaghị ọmụmụ ma e jiri ya tụnyere ọkụkọ na-achịkwa.Ndị na-ede akwụkwọ kwubiri na mgbanwe na mkpuchi glycoprotein-glycolipid gburugburu akpụkpọ ahụ spam na-ebelata ikike nke spam na fatịlaịza site na imebi sequestration nke spam na utero-vaginal junction, nke n'aka nke ya mụbara spam ọnwụ n'ihi ọsọ nke utero-ikpu junction, ma ọ dịghị emetụta spam na akwa.
Na turkey Bakst na Bauchan 11 chọtara obere vesicles na akpụkpọ anụ dị na lumen nke SST wee hụ na ụfọdụ n'ime granules ndị a jikọtara ya na akpụkpọ anụ spam.Ndị na-ede akwụkwọ na-atụ aro na mmekọrịta ndị a nwere ike inye aka na nchekwa ogologo oge nke spermatozoa na SST.Otú ọ dị, ndị nchọpụta akọwapụtaghị isi iyi nke ihe ndị a, ma ọ bụ mkpụrụ ndụ epithelial CCT zoro ezo, nke usoro ọmụmụ nwoke na-emepụta na zoro ezo, ma ọ bụ na-emepụta site na spam n'onwe ya.Ọzọkwa, ihe ndị a na-ahụ maka agglutination.Grützner et al27 kọrọ na mkpụrụ ndụ epididymal epithelial na-emepụta ma na-ezobe protein a kapịrị ọnụ nke achọrọ maka ịmepụta traktị seminal nke otu pore.Ndị ode akwụkwọ na-akọkwa na mgbasa nke ùkwù ndị a dabere na mmekọrịta nke protein epididymal.Nixon et al28 chọpụtara na adnexa na-ezobe protein, acidic cysteine-rich osteonectin;SPARC na-etinye aka n'ịmepụta sperm tufts na echidnas dị mkpụmkpụ na platypuses.A na-ejikọta mkpọsa nke osisi ndị a na mfu nke protein a.
N'ime ọmụmụ ihe ugbu a, nyocha nke ultrastructural site na iji microscopy eletrọn gosiri na spermatozoa na-arapara n'ọtụtụ nnukwu ihe.A na-eche na ihe ndị a bụ ihe kpatara agglutination nke na-eme ka ọ dị n'etiti na gburugburu isi ndị na-akwado ya, mana na obere mkpokọta na mpaghara ọdụ.Anyị na-eche na a na-ewepụ ihe agglutinating a site na usoro ọmụmụ nwoke (epididymis ma ọ bụ vas deferens) yana mmanụ, ebe ọ bụ na anyị na-ahụkarị semen na-ekewapụ na lymph na plasma seminal n'oge ejaculation.A kọwo na ka avian spermatozoa na-agafe na epididymis na vas deferens, ha na-enwe mgbanwe ndị metụtara maturation nke na-akwado ike ha ijikọ protein ma nweta plasma lemma glycoproteins.Ịnọgidesi ike na protein ndị a na akpụkpọ anụ sperm bi na SST na-egosi na ndị na-edozi ahụ nwere ike imetụta ịnweta nkwụsi ike nke sperm 30 ma chọpụta ọmụmụ 31 ha.Ahammad et al32 kọrọ na spermatozoa nwetara site n'akụkụ dị iche iche nke usoro ọmụmụ nwoke (site na testes na distal vas deferens) gosipụtara mmụba na-aga n'ihu na viability n'okpuru ọnọdụ nchekwa mmiri, n'agbanyeghị ọnọdụ nchekwa nchekwa, na viability na ọkụkọ na-abawanye na tubes fallopian mgbe insemination artificial.
Tufts ọkụkọ Sharkashi nwere njirimara na ọrụ dị iche iche karịa ụdị ndị ọzọ dị ka echidnas, platypuses, oke osisi, oke mgbada, na ezi pigs.Na ọkụkọ sharkasi, nhazi nke spermatozoa bundles belatara ọsọ igwu mmiri ha ma e jiri ya tụnyere otu spermatozoa.Otú ọ dị, ùkwù ndị a mụbara pasent nke rheologically nti spermatozoa ma mụbaa ikike nke spermatozoa iji mee ka onwe ha guzosie ike na gburugburu ebe dị ike.Ya mere, nsonaazụ anyị kwadoro aro gara aga na sperm agglutination na SST jikọtara ya na nchekwa spam ogologo oge.Anyị na-echekwa na mmasị nke spam na-etolite tufts nwere ike ịchịkwa ọnụ ọgụgụ spam na-efunahụ na SST, nke nwere ike gbanwee nsonaazụ asọmpi spam.Dị ka echiche a si dị, spermatozoa nwere ikike agglutination dị ala na-ahapụ SST mbụ, ebe spermatozoa nwere ikike agglutination dị elu na-emepụta ọtụtụ n'ime ụmụ.Nhazi nke ngwugwu sperm otu-pore bara uru ma na-emetụta oke nne na nna na nwa, mana na-eji usoro dị iche.Na echidnas na platypuses, a na-ahazi spermatozoa n'otu n'otu na ibe ya iji mee ka ọ na-aga n'ihu n'ibe ọkụ.Ngwunye echidnas na-aga ihe dị ka ugboro atọ ngwa ngwa karịa otu spermatozoa.A kwenyere na nguzobe sperm tufts dị otú ahụ na echidnas bụ mgbanwe evolushọn iji nọgide na-achị, ebe ọ bụ na ụmụ nwanyị na-enwe mmekọahụ na-enwekarị mmekọahụ na ọtụtụ ụmụ nwoke.Ya mere, spermatozoa sitere na ejaculates dị iche iche na-asọ mpi nke ukwuu maka fatịlaịza nke akwa.
Agglutinated spermatozoa nke sharkasi ọkụkọ dị mfe iji anya nke uche na-eji usoro iche microscopy, nke a na-ewere na ọ bara uru n'ihi na ọ na-enye ohere dị mfe ọmụmụ nke omume spermatozoa in vitro.Usoro nke sperm tuft guzobe akwalite mmeputakwa na sharkasi ọkụkọ dịkwa iche na nke a na-ahụ na ụfọdụ placental mammals na-anọchi anya sperm omume imekọ ihe ọnụ dị ka osisi òké, ebe ụfọdụ spermatozoa eru nsen, na-enyere ndị ọzọ metụtara ya iru na imebi ha àkwá.iji gosi onwe gị.omume ime mmụọ.Ime onwe 34. Ihe atụ ọzọ nke omume imekọ ihe ọnụ na spermatozoa ka ahụrụ n'ime ụmụ oke mgbada, ebe spermatozoa nwere ike ịchọpụta ma jikọta ya na spermatozoa kacha emetụta mkpụrụ ndụ ihe nketa ma mepụta otu ndị na-emekọ ihe ọnụ iji mee ka ọsọ ha dịkwuo elu ma e jiri ya tụnyere spermatozoa35 na-enweghị njikọ.
Nsonaazụ e nwetara na ọmụmụ ihe a adịghị emegide ozizi Foman nke nchekwa ogologo oge nke spermatozoa na SWS.Ndị nchọpụta na-akọ na mkpụrụ ndụ sperm na-aga n'ihu na-aga n'ihu na ntinye nke mkpụrụ ndụ epithelial na-ekpuchi SST ruo ogologo oge, na mgbe oge ụfọdụ gasịrị, ụlọ ahịa ike nke mkpụrụ ndụ sperm na-agwụ, na-eme ka mbelata ọsọ ọsọ, nke na-enye ohere ịchụpụ obere ihe dị arọ molecular.ike nke spermatozoa na eruba nke mmiri si lumen nke SST oghere nke fallopian tube.N'ime ọmụmụ ihe ugbu a, anyị hụrụ na ọkara nke otu spam na-egosi ike igwu mmiri megide mmiri na-asọ asọ, na nchikota ha na ngwugwu na-eme ka ha nwee ike igosi rheology dị mma.Ọzọkwa, data anyị kwekọrọ na nke Matsuzaki et al.1 onye kọrọ na ụbara secretion lactate na SST nwere ike igbochi sperm motility bi.Otú ọ dị, nsonaazụ anyị na-akọwa nguzobe nke sperm motile ligaments na omume ha rheological na ọnụnọ nke gburugburu ebe obibi dị ike n'ime microchannel na mgbalị iji kọwaa àgwà ha na SST.Nnyocha n'ọdịnihu nwere ike ilekwasị anya n'ịchọpụta ihe mejupụtara kemịkalụ na mmalite nke onye na-ahụ maka agglutinating, nke ga-enyere ndị na-eme nchọpụta aka ịmepụta ụzọ ọhụrụ iji chekwaa mmiri mmiri ma mee ka oge ọmụmụ dịkwuo elu.
A họpụtara sharkasi nwoke gbara olu gba ọtọ dị izu iri na ise dị afọ iri atọ ka ọ bụrụ ndị na-enye onyinye spam na ọmụmụ ihe.A zụlitere nnụnụ ndị a na Research Poultry Farm nke Ngalaba Agriculture, Mahadum Ashit, Ashit Governorate, Egypt.A na-edobe nnụnụ n'otu n'otu n'otu n'otu (30 x 40 x 40 cm), dabere na mmemme ọkụ (awa 16 nke ìhè na awa 8 nke ọchịchịrị) ma nye nri nwere 160 g nke protein crude, 2800 kcal nke ike metabolizable, 35 g nke calcium ọ bụla.5 grams nke phosphorus dị kwa kilogram nri.
Dị ka data 36, ​​37 si kwuo, a na-anakọta mkpụrụ n'aka ụmụ nwoke site na ịhịa aka n'ahụ afọ.Ngụkọta nke 45 semen sample ka a nakọtara site na mmadụ iri na ise n'ime ụbọchị 3.A na-agbaze Semen (n = 15 / ụbọchị) ozugbo 1: 1 (v: v) na Belsville Poultry Semen Diluent, nke nwere 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) na H2O (100 ml), pH = 7, 5, osmolarity 333 mOsm.A na-ebu ụzọ nyochaa ihe nlele nke mmanụ aṅụ n'okpuru microscope dị ọkụ iji hụ na mma mmiri dị mma (mmiri) wee chekwaa ya na mmiri ịsa ahụ na 37 Celsius C ruo mgbe ejiri ya n'ime ọkara elekere mgbe nchịkọta.
A na-akọwa kinematics na rheology nke spermatozoa site na iji usoro nke ngwaọrụ microfluidic.A na-agbanyekwa ihe nlele nke semen na 1: 40 na Beltsville Avian Semen Diluent, nke a na-ebunye n'ime ngwaọrụ microfluidic (lee n'okpuru ebe a), na usoro kinetic na-ekpebi site na iji usoro nyocha nke Semen Computerized (CASA) nke emebere na mbụ maka njirimara microfluidics.na ngagharị nke spermatozoa na mgbasa ozi mmiri (Department of Mechanical Engineering, Faculty of Engineering, Assiut University, Egypt).Enwere ike ibudata ngwa mgbakwunye ahụ na: http://www.assiutmicrofluidics.com/research/casa39.A tụrụ usoro ịgba ọsọ (VCL, μm/s), ọsọ ahịrị ahịrị (VSL, μm/s) na nkezi ọsọ trajectory (VAP, μm/s).Ewere vidiyo nke spermatozoa site na iji microscope nke Optika XDS-3 ntụgharị (nwere ebumnuche 40x) jikọtara na igwefoto Tucson ISH1000 na 30fps maka 3 s.Jiri ngwa CASA mụọ opekata mpe mpaghara atọ yana trajectories sperm 500 kwa nlele.Ejiri CASA arụrụ n'ụlọ hazie vidiyo e dekọrọ.Nkọwa nke motility na nkwụnye CASA dabeere na ọsọ igwu mmiri nke spam ma e jiri ya tụnyere ọnụ ọgụgụ na-asọpụta, ma ọ dịghị agụnye ihe ndị ọzọ dị ka mmegharị n'akụkụ n'akụkụ, n'ihi na achọpụtara na nke a bụ ihe a pụrụ ịdabere na ya na mmiri na-asọ.A na-akọwa mmegharị nke rheological dị ka mmegharị nke mkpụrụ ndụ sperm megide ntụziaka nke mmiri na-asọ.A na-ekewa spermatozoa na akụrụngwa rheological site na ọnụ ọgụgụ nke spermatozoa motile;spermatozoa ndị nọ na-ezu ike na spermatozoa na-emegharị ahụ na-ewepụ na ọnụ ọgụgụ ahụ.
E nwetara kemịkalụ niile ejirila n'aka Elgomhoria Pharmaceuticals (Cairo, Egypt) ọ gwụla ma amataghị ya.Emepụtara ngwaọrụ a dịka El-sherry et al kọwara.40 nwere mgbanwe ụfọdụ.Ihe eji emepụta microchannel gụnyere efere iko (Howard Glass, Worcester, MA), SU-8-25 negative resist (MicroChem, Newton, CA), mmanya diacetone (Sigma Aldrich, Steinheim, Germany), na polyacetone.-184, Dow Corning, Midland, Michigan).A na-emepụta microchannel site na iji lithography dị nro.Nke mbụ, a na-ebipụta ihe mkpuchi ihu doro anya nke nwere ụkpụrụ microchannel chọrọ na ngwa nbipute dị elu (Prismatic, Cairo, Egypt na Pacific Arts and Design, Markham, ON).E ji efere ugegbe mee ndị nna ukwu ahụ dị ka ihe eji eme ihe.A na-ehicha efere ndị ahụ na acetone, isopropanol na mmiri deionized wee kpuchie ya na 20 µm oyi akwa SU8-25 site na mkpuchi mkpuchi (3000 rpm, 1 min).A na-akpọzi akwa SU-8 nke ọma (65°C, 2 min na 95°C, 10 min) wee kpughee ya na radieshon UV maka 50 s.Ime ihe ngosi mgbe emechara na 65°C na 95°C maka 1 min na 4 min ka crosslink kpughere SU-8 layers, sochiri mmepe na mmanya diacetone maka nkeji 6.5.Richaa waffles siri ike (200 Celsius C maka 15 min) iji mee ka oyi akwa SU-8 sie ike.
A kwadebere PDMS site na ịgwakọta monomer na hardener na nha dị arọ nke 10: 1, wee kpochaa ya na onye na-ekpo ọkụ na-ekpo ọkụ ma wụsa ya na SU-8 isi etiti.A gwọrọ PDMS na oven (120 ° C, 30 min), mgbe ahụ, a na-ebipụ ọwa ndị ahụ, kewapụ ya na nna ukwu, ma gbanye ya ka ọ kwe ka ejikọta tubes na ntinye na ntinye nke microchannel.N'ikpeazụ, PDMS microchannels na-adịgide adịgide na mmịfe microscope site na iji ihe nrụpụta corona nwere ike ibugharị (Electro-Technical Products, Chicago, IL) dị ka akọwara ebe ọzọ.Microchannel eji na ọmụmụ ihe a tụrụ 200µm × 20 µm (W × H) yana ogologo 3.6 cm.
A na-enweta mmiri mmiri nke na-ebute site na nrụgide hydrostatic n'ime microchannel site n'ịkwado ọkwa mmiri dị na mmiri ntinye n'elu ihe dị elu dị iche Δh39 na ebe nchekwa mmiri (Fig 1).
ebe f bụ ọnụọgụ nke esemokwu, akọwapụtara dị ka f = C / Re maka laminar na-asọba na ọwa akụkụ anọ, ebe C bụ ihe na-adịgide adịgide dabere na akụkụ nke ọwa ahụ, L bụ ogologo microchannel, Vav bụ nkezi ọsọ n'ime microchannel, Dh bụ hydraulic dayameta nke ọwa, g - acceleration of gravity.N'iji nha nha a, enwere ike gbakọọ ọsọ ọwa nkezi site na iji nhata ndị a: