izvo maorganelle maviri asingakwanise kuparadzaniswa zviri nyore ne centrifugation

Kuparadzana Kwemasero: Kukunda Dambudziko Rekuparadzanisa Nuclei neMitochondria

Nhanganyaya:

Kugadzira centrifugation inzira ine simba inoshandiswa mu cellular ne molecular biology kuparadzanisa nekuparadzanisa organelles dzakasiyana kubva mumusanganiswa wakasiyana. Maitiro aya akavakirwa panheyo ye differential sedimentation, uko organelles dzakakora dzinogara pasi pechubhu pasi pe centrifugal force. Zvisinei, kune zvishoma zvisingabatanidzwi pamutemo uyu, uye organelles mbiri dzisingaparadzanise nyore ne centrifugation i nuclei ne mitochondria. Muchinyorwa chino, tichaongorora matambudziko anobatanidzwa mukuparadzanisa organelles idzi mbiri dzakakosha, kukosha kwekucheneswa kwadzo pamwe chete mune zvimwe zviedzo, uye dzimwe nzira dzekukunda dambudziko iri.

1. Kuomarara kweNuclei neMitochondria:

Nuclei, inowanzonzi nzvimbo yekuraira yesero, ndiyo inochengeta zvinhu zvemajini uye inodzora kuratidzwa kwemajini. Kune rumwe rutivi, mitochondria isimba guru rinogadzira simba muchimiro cheadenosine triphosphate (ATP). Nekuda kwebasa rayo rakasiyana uye hunhu hwayo hwakasiyana, kuparadzanisa maorganelle maviri aya kunogona kukonzera dambudziko guru.

2. Kukosha kwekuchenesa pamwe chete:

Kunyange zvazvo zvingava zvakanaka kuchenesa nhengo dzemuviri kuti zviitwe bvunzo dzakatarwa, pane dzimwe nguva apo kucheneswa pamwe chete kwe nuclei ne mitochondria kunonyanya kukosha. Semuenzaniso, pakudzidza mutemo wekugadzirwa kwemajini mukupindura shanduko mukugadzirwa kwemitochondrial, zvinova zvakakosha kuongorora kukanganisa kwekusashanda zvakanaka kwe mitochondrial pakugadzirwa kwemajini enyukireya. Muzviitiko zvakadaro, kuparadzaniswa kwemajini kubva ku mitochondria kwaizotungamira mukuongorora kusina kukwana uye kududzira kusina kururama.

3. Miganhu yeConventional Centrifugation:

Nzira dzechinyakare dzekushandisa centrifugation dzinoshandisa differential centrifugation, iyo inosanganisira nhevedzano yematanho ekushandisa centrifugation nekumhanya kwakasiyana kuti iparadzanise organelles nehukuru uye density. Zvinosuwisa kuti nuclei ne mitochondria zvine densities dzinopindirana uye hunhu hwakafanana hwe sedimentation, zvichiita kuti zvisave nyore kuzviparadzanisa uchishandisa nzira iyi chete. Izvi zvinonyanya kukonzerwa nehuwandu hwakawanda hwemafuta mu organelles dzese, zvichiita kuti dzifanane uye zvichikonzera co-sedimentation panguva ye centrifugation.

4. Dzimwe Nzira Dzekuzviparadzanisa:

Kuti vakunde zvipingamupinyi zve centrifugation, vaongorori vakagadzira dzimwe nzira dzekuparadzanisa nuclei ne mitochondria. Imwe nzira yakadaro ndeye density gradient centrifugation, iyo inoshandisa density gradient medium kugadzira continuous gradient. Nekuisa homogenate pamusoro pe gradient nekuibatanidza, organelles dzinogona kupatsanurwa zvichienderana nekudzikama kwadzo, zvichibvumira kupatsanurwa kwakanaka kwe nuclei ne mitochondria. Zvisinei, pasinei nekupatsanurwa kwakagadziridzwa, nzira iyi inogona kutora nguva yakareba uye yakaoma kuita.

5. Nzira Dzepamusoro: Magnetic Beads uye Fluorescent Tagging:

Mumakore achangopfuura, matekiniki epamusoro-soro anosanganisira magineti beads uye fluorescent tagging akakurumbira pakupatsanurwa kwe subcellular. Magineti beads akaputirwa nema specific antibodies anotarisa mapuroteni ari pamusoro pe nuclei kana mitochondria anogona kushandiswa kuparadzanisa organelle imwe kubva kune imwe. Uyezve, nekushandisa madhayi chaiwo e fluorescent akanangana ne nuclei kana mitochondria, organelles dzinogona kurongwa uchishandisa specialized flow cytometers kana fluorescence-activated cell sorting (FACS) machines. Aya matekiniki epamusoro anopa kururama kusati kwamboitika uye kushanda zvakanaka mukupatsanurwa kwe organelles idzi.

Mhedziso:

Kuparadzanisa nuclei ne mitochondria idambudziko nekuda kwekuenzana kwadzo uye huwandu hwadzo hwemafuta. Kunyange hazvo nzira dzechinyakare dze centrifugation dzichinetseka kuwana kupatsanurwa kwakakwana, matekiniki epamusoro akadai se density gradient centrifugation, magnetic bead-based isolation, uye fluorescent tagging zvakabuda sezvishandiso zvine simba zvekukunda zvipingamupinyi izvi. Nzira idzi itsva hadzingogadzirisi kushanda zvakanaka uye kururama kwekupatsanurwa kwe organelle asiwo dzinogonesa kuongororwa kwakazara kwebasa radzo uye kudyidzana kwadzo, zvichivhura nzira itsva dzekutsvaga kwemasero ne molecular.