Pehea e hoʻokaʻawale ai i nā lysosomes mai nā peroxisomes centrifugation
Hoʻolauna i nā Lysosomes a me nā Peroxisomes
ʻO Lysosomes a me peroxisomes ʻelua mau organelles cell kaʻawale me nā hana like ʻole e pāʻani ana i nā kuleana koʻikoʻi i ka metabolism cellular a me ka homeostasis. ʻO Lysosomes nā organelles i hoʻopaʻa ʻia i ka membrane i piha i nā enzymes digestive kuleana no ka wāwahi ʻana i nā mea ʻōpala cellular, nā ʻāpana haole, a me nā pathogens. ʻO nā peroxisomes, ma ka ʻaoʻao ʻē aʻe, pili i nā hopena metabolic like ʻole, ʻoiai nā mea e pili ana i ka detoxification a me ka metabolism lipid. No ke aʻo ʻana i kēia mau organelles ma ke kaʻawale, ʻo kahi ʻano hana i hoʻohana pinepine ʻia ʻo ka centrifugation. Ma kēia ʻatikala, e ʻimi mākou pehea e hiki ai ke hoʻohana ʻia ka centrifugation e hoʻokaʻawale pono i nā lysosomes mai nā peroxisomes.
Nā Kumumanaʻo a me nā ʻAno Hana o ka Centrifugation
He ʻano hana ikaika ka centrifugation i hoʻohana ʻia e hoʻokaʻawale i nā ʻāpana like ʻole o kahi hui heterogeneous e pili ana i ko lākou nui, density, a me ke ʻano. Hana ia ma nā kumumanaʻo o ka sedimentation, kahi e noho wikiwiki ai nā ʻāpana kaumaha ma lalo o ka mana o ka ikaika centrifugal. No ka hoʻokaʻawale ʻana i nā lysosomes mai nā peroxisomes, hiki ke hoʻohana ʻia nā ʻano hana centrifugation like ʻole, e like me ka differential centrifugation a me ka density gradient centrifugation.
ʻOkoʻa Centrifugation: Ke unuhi ʻana i nā Lysosomes a me nā Peroxisomes
ʻO ka centrifugation ʻokoʻa e pili ana i kahi moʻo o nā ʻanuʻu centrifugation ma nā wikiwiki a me nā lōʻihi like ʻole e hoʻokaʻawale i nā organelles ma muli o ko lākou nui. Eia pehea e hiki ai ke hoʻohana ʻia e unuhi i nā lysosomes a me nā peroxisomes:
KaʻAnuʻu Hana 1: Homogenization
ʻO ka hana mua, ʻo ia ka wāwahi mālie i nā ʻāpana a hoʻokuʻu i kā lākou mau mea me ka hoʻohana ʻana i kahi ʻano hana homogenization. Hiki ke hoʻokō ʻia kēia ma ka sonicating a i ʻole ka hoʻohana ʻana i kahi homogenizer kūikawā. ʻO ka hui hopena i kapa ʻia ʻo ka homogenate.
KaʻAnuʻu Hana 2: Centrifugation Haʻahaʻa-Wikiwiki
A laila hoʻokomo ʻia ka homogenate i ka centrifugation wikiwiki haʻahaʻa (ma kahi o 1,000-2,000 xg) no kahi manawa pōkole. Kōkua kēia hana i ka hoʻokaʻawale ʻana i nā ʻōpala cell nui a me ka hapa nuklea mai ke koena o nā organelles i loko o ka homogenate.
KaʻAnuʻu Hana 3: Centrifugation Waena-Wikiwiki
ʻO ka supernatant i loaʻa mai ke kaʻina centrifugation wikiwiki haʻahaʻa e hoʻokomo ʻia i kahi centrifugation wikiwiki waena (ma kahi o 10,000 xg) i nā organelles kaumaha e like me nā lysosomes a me nā peroxisomes. ʻO ka hopena o kēia kaʻina hana he pellet e loaʻa ana kēia mau organelles.
KaʻAnuʻu Hana 4: Holoi a hoʻokuʻu hou
Ua holoi ʻia ka pellet i loaʻa ma ke kaʻina hana ma mua me kahi hopena buffered kūpono a laila hoʻokuʻu hou ʻia e loaʻa kahi ʻāpana lysosome a me peroxisome i hoʻomaʻemaʻe ʻia.
ʻO ka Density Gradient Centrifugation: Hoʻomaʻemaʻe i ka hoʻokaʻawale ʻana
ʻO ka centrifugation gradient density kahi ʻenehana centrifugation holomua e hiki ai ke hoʻomaʻemaʻe hou aʻe a hoʻomaikaʻi i nā lysosomes a me nā peroxisomes. Ma kēia ʻenehana, hoʻokumu ʻia kahi gradient density me ka hoʻohana ʻana i kahi hopena gradient-forming e like me ka sucrose a i ʻole iodixanol. Eia pehea e hiki ai ke hoʻopili ʻia:
KaʻAnuʻu Hana 1: Hoʻomākaukau ʻana o ka Density Gradient
Hoʻomākaukau ʻia kahi medium gradient density, e like me ka sucrose, i loko o kahi paipu centrifuge, me nā density like ʻole mai luna a lalo a i ʻole lalo a luna. Ua koho pono ʻia ke ʻano a me ka density o ka medium gradient, e hōʻoia ana he māmā ia ma mua o nā peroxisomes akā ʻoi aku ka denser ma mua o nā lysosomes.
KaʻAnuʻu Hana 2: Ka Hoʻohui ʻana i ka Laʻana
Ua hoʻomoe pono ʻia ka ʻāpana lysosome a me peroxisome i hoʻoulu hou ʻia i loaʻa mai ka centrifugation differential ma luna o ka medium gradient density.
KaʻAnuʻu Hana 3: Centrifugation
A laila hoʻokomo ʻia ka ʻōmole i ka centrifugation wikiwiki (ultracentrifugation) e hoʻokaʻawale i nā lysosomes a me nā peroxisomes ma muli o ko lākou mau densities buoyant. I ka wā o ka centrifugation, neʻe nā organelles ma waena o ka medium a hiki i kahi ʻāpana kahi e kūlike ai ko lākou density buoyant me ka density o ka medium a puni, e hana ana i nā hui like ʻole.
Ka Nānā ʻana a me nā Noi Hou Aʻe
Ma hope o ka hoʻokaʻawale kūleʻa ʻana, hiki ke kālailai hou ʻia nā ʻāpana lysosome a me peroxisome i loaʻa me ka hoʻohana ʻana i nā ʻano hana like ʻole e loiloi i ko lākou maʻemaʻe, ka nui, ka haku mele ʻana, a me nā waiwai hana. Hiki i nā ʻano hana e like me ka microscopy electron, immunocytochemistry, a me nā hoʻāʻo hana enzyme ke hāʻawi i nā ʻike waiwai i nā ʻāpana i hoʻokaʻawale ʻia, e hōʻoia ana i ko lākou ʻano a me ka maʻemaʻe.
Ma waho aʻe o nā kumu noiʻi, ʻo ka hiki ke hoʻokaʻawale i nā lysosomes a me nā peroxisomes ma o ka centrifugation he mau noi i nā diagnostics lapaʻau, ka hoʻomohala ʻana i nā lāʻau lapaʻau, a me nā hana therapeutic. Ma ka hoʻomaopopo maikaʻi ʻana i nā hana o kēia mau organelles, hiki i nā kānaka ʻepekema ke ʻimi i nā hoʻolālā therapeutic hou e pili ana i nā maʻi lysosomal a i ʻole peroxisomal, e like me nā maʻi mālama lysosomal a i ʻole nā maʻi biogenesis peroxisome.
I ka hopena, ʻo ka hoʻokaʻawale ʻana o nā lysosomes mai nā peroxisomes me ka hoʻohana ʻana i nā ʻano hana centrifugation, e like me ka differential centrifugation a me ka density gradient centrifugation, he ala koʻikoʻi ia i ka noiʻi biology cell. Hiki i kēia mau ʻano hana i nā kānaka ʻepekema ke loaʻa nā ʻāpana i hoʻomaʻemaʻe ʻia o nā lysosomes a me nā peroxisomes, e kōkua ana i ka hoʻomaopopo ʻana i kā lākou kuleana pākahi, nā ala metabolic, a me nā hana therapeutic hiki ke hana no nā maʻi pili.
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