ukuthi yiziphi i-organelle ezimbili ezingenakuhlukaniswa kalula nge-centrifugation

Ukuhlukaniswa Kwamaseli Angaphansi: Ukunqoba Inselele Yokuhlukanisa Ama-Nuclei nama-Mitochondria

Isingeniso:

I-Centrifugation iyindlela enamandla esetshenziswa ku-cell kanye ne-molecular biology ukuhlukanisa nokuhlukanisa ama-organelle ahlukahlukene engxubeni ehlukahlukene. Le nqubo isekelwe esimisweni se-differential sedimentation, lapho ama-organelle asindayo ehlala phansi kwepayipi ngaphansi kwamandla e-centrifugal. Kodwa-ke, kunezimbalwa ezihlukile kulo mthetho, futhi ama-organelle amabili angahlukani kalula nge-centrifugation yi-nuclei kanye ne-mitochondria. Kulesi sihloko, sizohlola izinselele ezihilelekile ekuhlukaniseni la ma-organelle amabili abalulekile, ukubaluleka kokuhlanzwa kwawo ngokubambisana ekuhlolweni okuthile, kanye nezindlela ezihlukile zokunqoba lesi sithiyo.

1. Ubunzima be-Nuclei ne-Mitochondria:

I-Nuclei, evame ukubizwa ngokuthi isikhungo somyalo weseli, inesibopho sokugcina izinto zofuzo kanye nokulawula ukuvezwa kwezakhi zofuzo. Ngakolunye uhlangothi, i-mitochondria iyimithombo ebalulekile yamandla ekhiqiza amandla ngesimo se-adenosine triphosphate (ATP). Ngenxa yezindima zayo ezihlukile kanye nezici zayo ezihlukile zesakhiwo, ukuhlukanisa la ma-organelle amabili kungaba yinselele enkulu.

2. Ukubaluleka Kokuhlanzwa Kokubambisana:

Nakuba kungase kube yinto efiselekayo ukuhlanza ama-organelle ngamanye ukuze kwenziwe izivivinyo ezithile, kunezimo lapho ukuhlanzwa ngokubambisana kwama-nuclei kanye nama-mitochondria kubaluleke kakhulu. Isibonelo, lapho kufundwa ukulawulwa kokubonakaliswa kwezakhi zofuzo ngokuphendula izinguquko ekusetshenzisweni kwe-mitochondrial, kuba yinto ebalulekile ukuhlaziya umthelela wokungasebenzi kahle kwe-mitochondrial ekubonakalisweni kwezakhi zofuzo zenuzi. Ezimweni ezinjalo, ukuhlukaniswa kwama-nuclei kusuka kuma-mitochondria kungaholela ekuhlaziyweni okungaphelele kanye nokuchazwa okunganembile.

3. Imikhawulo Yokuvinjelwa Okuvamile Kokucisha Ugesi:

Izindlela zendabuko zokusebenzisa i-centrifugation zisebenzisa i-differential centrifugation, ehilela uchungechunge lwezinyathelo zokusebenzisa i-centrifugation ngesivinini esihlukahlukene ukuze kuhlukaniswe ama-organelle ngobukhulu nangobuningi bawo. Ngeshwa, ama-nuclei nama-mitochondria anobuningi obuhambisanayo kanye nezakhiwo ezifanayo zokusebenzisa i-sedimentation, okwenza kube nzima ukuwahlukanisa kusetshenziswa le ndlela yodwa. Lokhu kungenxa kakhulu yokuqukethwe okuphezulu kwamafutha kuzo zombili i-organelles, okuholela ekubutheni kwazo okufanayo futhi okuholela ekubuthweni kwe-sedimentation ngesikhathi sokusebenzisa i-centrifugation.

4. Ezinye Izindlela Zokuzihlukanisa:

Ukuze banqobe ukulinganiselwa kwe-centrifugation, abacwaningi bathuthukise amasu ahlukile okuhlukaniswa kwe-nuclei ne-mitochondria. Enye indlela enjalo i-density gradient centrifugation, esebenzisa i-density gradient medium ukwakha i-gradient eqhubekayo. Ngokubeka ngokucophelela i-homogenate phezu kwe-gradient nokuyi-centrifugation, ama-organelle angahlukaniswa ngokusekelwe ekubutheni kwawo, okuvumela ukuhlukaniswa kwe-nuclei ne-mitochondria okusebenza kahle. Kodwa-ke, naphezu kokuhlukaniswa okuthuthukisiwe, le ndlela ingathatha isikhathi eside futhi kunzima kakhulu ukuyenza.

5. Izindlela Ezithuthukisiwe: Ubuhlalu Obunemaginethi kanye Nokuthegiswa Okukhanyayo:

Eminyakeni yamuva nje, amasu athuthukile ahilela ubuhlalu obunamandla kanye nokumaka okukhanyayo athole ukuthandwa kokuhlukaniswa kwe-subcellular. Ubuhlalu obunamandla obumbozwe ngama-antibodies athile aqondisa amaprotheni aphezulu akhona kuma-nuclei noma kuma-mitochondria bungasetshenziswa ukuhlukanisa i-organelle eyodwa kwenye ngokukhetha. Ngaphezu kwalokho, ngokusebenzisa udayi oluthile olukhanyayo oluqondiswe kuma-nuclei noma kuma-mitochondria, ama-organelle angahlelwa kusetshenziswa ama-flow cytometers akhethekile noma imishini yokuhlunga amaseli asebenze nge-fluorescence (FACS). Lobu buchwepheshe obusezingeni eliphezulu bunikeza ukunemba nokusebenza kahle okungakaze kubonwe ekuhlukanisweni kwala ma-organelle.

Isiphetho:

Ukuhlukanisa ama-nuclei nama-mitochondria kuyinselele ngenxa yokufana kwawo kobuningi kanye nobuningi bamafutha. Ngenkathi izindlela zendabuko zokuhlanganisa ama-centrifugation zilwela ukufeza ukuhlukaniswa okwanele, amasu athuthukile njenge-density gradient centrifugation, i-magnetic bead-based isolation, kanye ne-fluorescent tagging avele njengamathuluzi anamandla okunqoba lezi zithiyo. Lezi zindlela ezintsha azigcini nje ngokuthuthukisa ukusebenza kahle kanye nokunemba kokuhlukaniswa kwama-organelle kodwa futhi zivumela ukuhlaziywa okuphelele kwemisebenzi yazo kanye nokusebenzisana, zivula izindlela ezintsha zocwaningo lwamaseli nama-molecule.