he aha ngā whekau e rua kāore e taea te wehewehe ngāwari mā te centrifugation
Te Wehenga Pūtau-iti: Te wikitoria i te wero o te wehewehe i ngā karihi me ngā mitochondria
Kupu Whakataki:
He tikanga kaha te centrifugation e whakamahia ana i roto i te koiora pūtau me te ngota ngota hei wehe me te wehe i ngā momo okana mai i te ranunga kanorau. Ko te tukanga e hangai ana ki te kaupapa o te whakatōtōtanga rerekētanga, ko te wāhi e tau ai ngā okana taumaha ki raro i te ngongo i raro i te kaha centrifugation. Heoi, he iti noa ngā tūponotanga ki tēnei ture, ā, e rua ngā okana kāore e ngāwari te wehe mā te centrifugation ko ngā karihi me ngā mitochondria. I roto i tēnei tuhinga, ka tūhuratia e mātou ngā wero e pā ana ki te wehewehe i ēnei okana nui e rua, te hiranga o tā rātou purenga tahi i roto i ētahi whakamātautau, me ngā tikanga rerekē hei wikitoria i tēnei arai.
1. Te Uauatanga o ngā Nuclei me te Mitochondria:
Ko ngā karihi, e mōhiotia whānuitia ana ko te pokapū whakahaere o te pūtau, ko rātou te kawenga mō te pupuri i ngā rauemi ira me te whakahaere i te whakaaturanga ira. Ko ngā mitochondria ia, he pūngao nui e whakaputa ana i te pūngao i roto i te ahua o te adenosine triphosphate (ATP). Nā ō rātou mahi motuhake me ngā āhuatanga hanganga ahurei, he uaua rawa te wehewehe i ēnei whekau e rua.
2. Te Hiranga o te Whakamaarama Ngātahi:
Ahakoa he mea pai pea te pure i ngā whekau takitahi mō ngā whakamātautau motuhake, tera ētahi wā he mea nui te purenga tahi o ngā karihi me ngā mitochondria. Hei tauira, i te wā e ako ana i te whakahaere i te whakaaturanga ira hei whakautu i ngā huringa o te pākia o te mitochondria, ka noho hei mea nui te tātari i te pānga o te ngoikore o te mitochondria ki te whakaaturanga ira karihi. I roto i ēnei take, ko te wehenga o ngā karihi mai i ngā mitochondria ka arahi ki te tātaritanga kāore i oti me ngā whakamaoritanga hē.
3. Ngā Here o te Whakapūtanga Tikanga:
Ko ngā tikanga tuku iho o te centrifugation he whakamahi i te centrifugation rerekē, arā, he raupapa o ngā mahi centrifugation i ngā tere rerekē hei wehewehe i ngā organelles mā te rahi me te mātotoru. Heoi anō, he rite ngā mātotorutanga o ngā karihi me ngā mitochondria, ā, he rite tonu ngā āhuatanga o te whakapūpū, ā, he mea e kore e taea te wehewehe i a rātou mā te whakamahi i tēnei huarahi anake. Ko te take nui o tēnei ko te nui o te ngako i roto i ngā organelles e rua, e hua ake ana i te rite o te pūmautanga, ā, ka hua ake he whakapūpūtanga ngātahi i te wā o te centrifugation.
4. Ngā Tikanga Rerekē mō te Wehenga:
Hei whakakore i ngā herenga o te centrifugation, kua whakawhanakehia e ngā kairangahau ētahi atu tikanga hei wehewehe i ngā karihi me ngā mitochondria. Ko tētahi o aua tikanga ko te centrifugation gradient density, e whakamahi ana i tētahi reo gradient density hei hanga i tētahi gradient tonu. Mā te whakatakoto āta i te homogenate ki runga i te gradient me te centrifugation, ka taea te wehewehe i ngā organelles i runga i tō rātou pūmau, ka taea ai te wehewehe i ngā karihi me ngā mitochondria me te whai hua. Heoi, ahakoa te pai ake o te wehenga, ka roa ake pea tēnei tikanga, ā, he uaua ake te mahi.
5. Ngā Huarahi Matatau: Ngā Pīrere Aukume me te Tohu Hikoi:
I ngā tau tata nei, kua rongonui haere ngā tikanga matatau e whakamahi ana i ngā pirepire aukume me te tohu fluorescence mō te wehenga pūtau-iti. Ka taea te whakamahi i ngā pirepire aukume kua pania ki ngā paturopi motuhake e aro ana ki ngā pūmua mata kei runga i ngā karihi, i ngā mitochondria rānei, hei wehe motuhake i tētahi okana mai i tētahi atu. Hei tāpiri, mā te whakamahi i ngā tae fluorescence motuhake e aro ana ki ngā karihi, ki ngā mitochondria rānei, ka taea te whakarōpū i ngā okana mā te whakamahi i ngā cytometer rere motuhake, i ngā mīhini whakarōpū pūtau fluorescence-activated (FACS) rānei. Ka tukuna e ēnei hangarau hou te tika me te whai huatanga o mua i roto i te wehenga o ēnei okana.
Whakatau:
He wero te wehewehe i ngā karihi me ngā mitochondria nā te mea he rite te matotoru me te nui o te ngako. Ahakoa he uaua ki ngā tikanga centrifugation tuku iho te whakatutuki i te wehenga tika, kua puta ake ngā tikanga matatau pēnei i te centrifugation density gradient, te wehenga i runga i te bead-based magnetic, me te fluorescent tagging hei taputapu kaha mō te wikitoria i ēnei arai. Ehara i te mea ka whakapai ake ēnei tikanga auaha i te whai huatanga me te tika o te wehenga organelle engari ka taea hoki te tātari whānui i ā rātou mahi me ngā taunekeneke, ka whakatuwhera i ngā huarahi hou mō te rangahau pūtau me te ngota.
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