indlela yokuhlukanisa i-chloroplast ne-mitochondria ngokusebenzisa i-centrifugation

I-Centrifugation: Indlela Enamandla Yokuhlukanisa I-Chloroplast Ne-Mitochondria

Isingeniso:

I-Centrifugation iyindlela esetshenziswa kabanzi ku-molecular biology kanye ne-biochemistry evumela ukuhlukaniswa kwezingxenye ezahlukene zamaseli ngokusekelwe kusayizi wazo, ukuma, kanye nobuningi bazo. Lesi sihloko sigxila ekusetshenzisweni kwe-centrifugation ekuhlukaniseni ama-chloroplast kanye ne-mitochondria, ama-organelle amabili abalulekile atholakala kumaseli e-eukaryotic ahilelekile ekukhiqizweni kwamandla kanye nezinye izinqubo ezibalulekile zamaseli.

Indima Yama-Chloroplast Nama-Mitochondria Ezinqubweni Zamaseli

Ama-chloroplast, akhona kumaseli ezitshalo, anesibopho se-photosynthesis, inqubo eguqula amandla okukhanya abe amandla amakhemikhali agcinwe ngesimo se-glucose. Le nqubo ebalulekile yenza kube lula ukukhiqizwa komoya-mpilo futhi iyisisekelo sokusekela impilo eMhlabeni. Ama-Mitochondria, atholakala kokubili kumaseli ezitshalo nezilwane, avame ukubizwa ngokuthi yi-powerhouse yeseli. Anesibopho sokuphefumula kwamaseli, aguqula i-glucose ibe amandla angasetshenziswa yiseli ngesimo se-adenosine triphosphate (ATP).

Isidingo Sokuhlukanisa Ama-Chloroplast Nama-Mitochondria

Ukuqonda imisebenzi kanye nezici zama-chloroplast kanye nama-mitochondria kubalulekile kubacwaningi abahlose ukuthola ukuqonda ngezinqubo zamaseli nokuthuthukisa izindlela zokwelapha izifo ezahlukahlukene. Ukuhlukanisa la ma-organelles engxubeni eyinkimbinkimbi yezingxenye zamaseli kubalulekile ukuze kutholakale amasampula amsulwa okuhlaziywa okwengeziwe nokuhlolwa. Lokhu kuhlukaniswa kungenzeka nge-centrifugation.

Izisekelo Zokugqamisa I-Centrifugation

Ukushintshashintsha kuhilela ukusetshenziswa kwamandla e-centrifugal ukuhlukanisa izingxenye ngaphakathi kwesampula. Isampula ifakwa ku-centrifuge, umshini one-rotor ejikeleza ngesivinini esikhulu. Ngesikhathi sokujikeleza, amandla e-centrifugal abangela izingxenye ezixinene kakhulu ukuba zichitheke phansi, zakhe i-pellet, kuyilapho izingxenye ezingaxinene kakhulu zihlala ku-supernatant. Ijubane kanye nobude be-centrifuge, kanye nokukhethwa kwamashubhu e-centrifuge, kuyahlukahluka kuye ngokusetshenziswa okuthile kanye nokwakheka kwesampula.

I-Centrifugation Ehlukile Yokuhlukaniswa Kwe-Organelle

Indlela evame ukusetshenziswa kakhulu yokuhlukanisa ama-organelle i-differential centrifugation. Le ndlela isekelwe esimisweni sokuhlukanisa isampula ngokuyifaka ochungechungeni lwe-differential centrifugations ngesivinini esikhulayo kanye nobude besikhathi. Ngokulungisa amapharamitha e-centrifugation, abacwaningi bangahlukanisa ngempumelelo ama-chloroplast kanye nama-mitochondria kwezinye izingxenye zamaseli.

Umhlahlandlela Wesinyathelo Ngesinyathelo Wokuhlukanisa Ama-Chloroplast Nama-Mitochondria

Isinyathelo 1: Ukuhlanganiswa kwe-Homogenization

Isinyathelo sokuqala ekuhlukaniseni i-differential centrifugation ukuhlanganisa isampula ngendlela efanayo. Le nqubo ihilela ukuphula amaseli kancane bese kukhishwa okuqukethwe kwawo, okuvumela ukufinyelela kuma-organelle. Izindlela eziningana zingasetshenziswa ekuphazamiseni amaseli, njengokuphazamiseka kwemishini kanye nokwelashwa kwe-nitrogen ewuketshezi.

Isinyathelo 2: I-Differential Centrifugation

Ngemva kokuhlanganiswa kwe-homogenization, isampula ifakwa ochungechungeni lwe-differential centrifugations. I-centrifugation yokuqala yesivinini esiphansi (1000-2000 xg) isusa udoti omkhulu wamaseli namaseli angaphukile, ishiye i-mitochondria eluhlaza kanye nengxenye ye-chloroplast. Le ngxenye ibe isifakwa i-centrifugation yesivinini esiphezulu (10,000-15,000 xg), okuholela ekuhlukanisweni kwama-chloroplast kuma-mitochondria.

Isinyathelo 3: Ukuthuthukiswa Kobumsulwa

Ukuze kuhlanzwe kabanzi izingxenyana ze-chloroplast kanye ne-mitochondria ezitholiwe, kungenziwa izinyathelo ezengeziwe zokuhlanganisa i-centrifugation. Lokhu kuhilela ukuphinda kumiswe i-pellet etholwe ngemva kokuhlanganisa i-centrifugation ngakunye ku-buffer efanele bese kuphinda inqubo ngesivinini esikhulu kanye nobude besikhathi. Lokhu kuhlanganisa i-centrifugation okwengeziwe kuzosiza ekususeni noma yikuphi ukungcola okusele futhi kwandise ubumsulwa bezingxenyana zokugcina ze-organelle.

Isiphetho:

I-Centrifugation iyindlela eguquguqukayo eye yashintsha insimu yebhayoloji yamaseli futhi isibe ithuluzi elibalulekile lokuhlukaniswa kwezingxenye zamaseli. Ngokusebenzisa i-differential centrifugation, abacwaningi bangahlukanisa ngempumelelo ama-chloroplast nama-mitochondria, banikeze amasampula ahlanzekile abalulekile ukuze kuhlaziywe futhi kuhlolwe okwengeziwe. Amandla okuhlukanisa la ma-organelle avula iminyango yokufunda imisebenzi yawo ethile kanye nezindlela, okufaka isandla ekuqondeni kangcono izinqubo zamaseli kanye nokukhuthaza intuthuko emikhakheni eyahlukene yesayensi.