ʻAʻole hiki ke hoʻokaʻawale maʻalahi ʻia nā organelles ʻelua e ka centrifugation
Ka Hoʻokaʻawale ʻana o Subcellular: Ke Lanakila nei i ka Pilikia o ka Hoʻokaʻawale ʻana i nā Nuclei a me Mitochondria
Hoʻolauna:
He ʻano hana ikaika ka centrifugation i hoʻohana ʻia i loko o ke olaola cellular a me ka molekala e hoʻokaʻawale a hoʻokaʻawale i nā organelles like ʻole mai kahi hui heterogeneous. Hoʻokumu ʻia ke kaʻina hana ma ke kumumanaʻo o ka sedimentation differential, kahi e noho ai nā organelles kaumaha ma lalo o kahi paipu ma lalo o ka ikaika centrifugal. Eia nō naʻe, aia kekahi mau ʻokoʻa i kēia lula, a ʻelua mau organelles i hoʻokaʻawale maʻalahi ʻole ʻia e ka centrifugation ʻo ia nā nuclei a me nā mitochondria. Ma kēia ʻatikala, e ʻimi mākou i nā pilikia e pili ana i ka hoʻokaʻawale ʻana i kēia mau organelles koʻikoʻi ʻelua, ke koʻikoʻi o kā lākou hoʻomaʻemaʻe like ʻana i kekahi mau hoʻokolohua, a me nā ʻano hana ʻē aʻe e lanakila ai i kēia pilikia.
1. Ka Paʻakikī o nā Nuclei a me Mitochondria:
ʻO nā nuclei, i kapa pinepine ʻia ʻo ke kikowaena kauoha o ke kelepona, ke kuleana no ka mālama ʻana i nā mea genetic a me ka hoʻoponopono ʻana i ka hōʻike ʻana o nā gene. ʻO Mitochondria, ma ka ʻaoʻao ʻē aʻe, he mau hale mana koʻikoʻi e hoʻopuka i ka ikehu ma ke ʻano o adenosine triphosphate (ATP). Ma muli o kā lākou kuleana like ʻole a me nā ʻano hoʻonohonoho kū hoʻokahi, hiki i ka hoʻokaʻawale ʻana i kēia mau organelles ʻelua ke lilo i mea paʻakikī nui.
2. Ke Koʻikoʻi o ka Hoʻomaʻemaʻe Pū ʻana:
ʻOiai he mea makemake paha e hoʻomaʻemaʻe i nā organelles pākahi no nā hoʻokolohua kikoʻī, aia kekahi mau hihia kahi e koʻikoʻi ai ka hoʻomaʻemaʻe like ʻana o nā nuclei a me mitochondria. No ka laʻana, i ke aʻo ʻana i ka hoʻoponopono ʻana o ka gene i ka pane ʻana i nā loli i ka metabolism mitochondrial, lilo ia i mea koʻikoʻi e kālailai i ka hopena o ka hana ʻino o ka mitochondrial i ka hōʻike ʻana o ka gene nuklea. I ia mau hihia, ʻo ka hoʻokaʻawale ʻana o nā nuclei mai mitochondria e alakaʻi i ka nānā piha ʻole a me nā wehewehe pololei ʻole.
3. Nā Palena o ka Centrifugation Kuʻuna:
Hoʻohana nā ʻano centrifugation kuʻuna i ka centrifugation ʻokoʻa, kahi e pili ana i kahi moʻo o nā ʻanuʻu centrifugation ma nā wikiwiki like ʻole e hoʻokaʻawale i nā organelles ma ka nui a me ka nui. ʻO ka mea pōʻino, he mau nui like nā nuclei a me nā mitochondria a me nā waiwai sedimentation like, e aneane hiki ʻole ke hoʻokaʻawale iā lākou me ka hoʻohana ʻana i kēia ʻano hana wale nō. ʻO ke kumu nui o kēia ma muli o ka nui o ka lipid i loko o nā organelles ʻelua, e alakaʻi ana i ko lākou buoyancy like a me ka hopena i ka co-sedimentation i ka wā centrifugation.
4. Nā ʻAno Hana ʻē aʻe no ka Hoʻokaʻawale ʻana:
I mea e lanakila ai i nā palena o ka centrifugation, ua hoʻomohala nā mea noiʻi i nā ʻano hana ʻē aʻe no ka hoʻokaʻawale ʻana o nā nuclei a me mitochondria. ʻO kekahi o ia ʻano hana, ʻo ia ka density gradient centrifugation, kahi e hoʻohana ai i kahi medium density gradient e hana i kahi gradient hoʻomau. Ma ke kau pono ʻana i ka homogenate ma luna o ka gradient a me ka centrifuging ʻana, hiki ke hoʻokaʻawale ʻia nā organelles ma muli o ko lākou buoyancy, e ʻae ana i ka hoʻokaʻawale pono ʻana o nā nuclei a me mitochondria. Eia nō naʻe, ʻoiai ua hoʻomaikaʻi ʻia ka hoʻokaʻawale ʻana, hiki ke lōʻihi kēia ʻano hana a ʻoi aku ka paʻakikī o ka hana ʻana.
5. Nā ʻAno Holomua: Nā Pīpī Magnetic a me ka Hōʻailona Fluorescent:
I nā makahiki i hala iho nei, ua kaulana nā ʻano hana holomua e pili ana i nā momi magnetic a me ka fluorescent tagging no ka hoʻokaʻawale ʻana o nā subcellular. Hiki ke hoʻohana ʻia nā momi magnetic i uhi ʻia me nā antibodies kikoʻī e kuhikuhi ana i nā protein ʻili i loaʻa ma nā nuclei a i ʻole mitochondria e hoʻokaʻawale koho i kekahi organelle mai kekahi. Eia kekahi, ma ka hoʻohana ʻana i nā dyes fluorescent kikoʻī i kuhikuhi ʻia i nā nuclei a i ʻole mitochondria, hiki ke hoʻokaʻawale ʻia nā organelles me ka hoʻohana ʻana i nā cytometers kahe kūikawā a i ʻole nā mīkini hoʻokaʻawale cell fluorescence-activated (FACS). Hāʻawi kēia mau ʻenehana holomua i ka pololei a me ka pono i ʻike ʻole ʻia i ka hoʻokaʻawale ʻana o kēia mau organelles.
Hopena:
He paʻakikī ka hoʻokaʻawale ʻana i nā nuclei a me nā mitochondria ma muli o ko lākou ʻano like a me ke ʻano waiwai lipid. ʻOiai ke hakakā nei nā ʻano centrifugation kuʻuna e hoʻokō i ka hoʻokaʻawale kūpono, ua puka mai nā ʻano hana holomua e like me ka density gradient centrifugation, magnetic bead-based isolation, a me ka fluorescent tagging ma ke ʻano he mau mea hana ikaika no ka lanakila ʻana i kēia mau pilikia. ʻAʻole wale kēia mau ʻano hana hou e hoʻomaikaʻi i ka pono a me ka pololei o ka hoʻokaʻawale ʻana o nā organelle akā hiki nō hoʻi ke ʻae i ka nānā piha ʻana i kā lākou mau hana a me nā pilina, e wehe ana i nā ala hou no ka noiʻi cellular a me molecular.
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