Mokhoa oa ho arola li-lysosomes ho centrifugation ea peroxisomes

Selelekela ho Lysosomes le Peroxisomes

Li-lysosome le li-peroxisome ke li-organelle tse peli tse arohaneng tsa sele tse nang le mesebetsi e ikhethang e bapalang karolo ea bohlokoa metabolism ea sele le homeostasis. Li-lysosome ke li-organelle tse tlamiloeng ka lera tse tletseng li-enzyme tsa tšilo ea lijo tse ikarabellang bakeng sa ho qhaqha litšila tsa sele, likaroloana tse tsoang linaheng tse ling le likokoana-hloko. Ka lehlakoreng le leng, li-peroxisome li ameha liketsong tse fapaneng tsa metabolism, haholo-holo tse amanang le ho ntša chefo le metabolism ea lipid. Ho ithuta li-organelle tsena ka thoko, mokhoa o mong o sebelisoang haholo ke centrifugation. Sehloohong sena, re tla hlahloba kamoo centrifugation e ka sebelisoang kateng ho arola li-lysosome ho li-peroxisome ka katleho.

Melao-motheo le Mekhoa ea ho Centrifugation

Ho centrifugation ke mokhoa o matla o sebelisoang ho arola likarolo tse fapaneng tsa motsoako o fapaneng ho latela boholo ba tsona, bongata le sebopeho. E sebetsa ka melao-motheo ea ho sedimentation, moo likaroloana tse boima li lulang kapele tlas'a tšusumetso ea matla a centrifugal. Ho arola lysosomes ho peroxisomes, mekhoa e fapaneng ea ho centrifugation, joalo ka ho centrifugation e fapaneng le ho centrifugation ea density gradient, e ka sebelisoa.

Ho Centrifugation e Fapaneng: Ho Ntša Lysosomes le Peroxisomes

Ho arola dipharologanyo ho kenyeletsa letoto la mehato ya ho arola dipharologanyo ka lebelo le bolelele bo fapaneng ho arola di-organelle ho latela bongata ba tsona. Mona ke kamoo e ka sebediswang ho ntsha di-lysosome le di-peroxisome:

Mohato oa 1: Homogenization

Mohato oa pele ke ho qhaqha lisele ka bonolo le ho lokolla lintho tse ka hare ho tsona ho sebelisoa mokhoa oa ho kopanya lisele. Sena se ka finyelloa ka ho sebelisa sonicating kapa ho sebelisa homogenizer e khethehileng. Motsoako o hlahisoang o bitsoa homogenate.

Mohato oa 2: Ho Fokotsa ho Centrifugation ka Lebelo le Tlase

Homogenate ebe e kenngoa ka lebelo le tlase la centrifugation (hoo e ka bang 1,000-2,000 xg) ka nako e khuts'oane. Mohato ona o thusa ho arola masalla a maholo a sele le karolo ea nyutlelie ho tsoa ho li-organelle tse ling tse teng homogenate.

Mohato oa 3: Ho Centrifugation ea Lebelo le Mahareng

Supernatant e fumanoeng mohatong oa ho centrifugation ka lebelo le tlase e kenngoa ho centrifugation ka lebelo le mahareng (hoo e ka bang 10,000 xg) ho ea ho li-organelle tse boima tse kang lysosomes le peroxisomes. Mohato ona o fella ka pellet e nang le li-organelles tsena.

Mohato oa 4: Hlatsoa 'me u tsosolose

Pellet e fumanoeng mohatong o fetileng e hlatsuoa ka tharollo e loketseng e nang le buffer ebe e emisoa hape ho fumana karolo e hloekisitsoeng ea lysosome le peroxisome.

Ho Ntlafatsa Karohano ka Botenya ba Gradient: Ho Ntlafatsa Karohano

Ho centrifugation ha gradient e teteaneng ke mokhoa o tsoetseng pele oa ho centrifugation o lumellang ho hloekisoa le ho ntlafatsoa ho eketsehileng ha lysosomes le peroxisomes. Mokhoeng ona, gradient e teteaneng e thehoa ho sebelisoa tharollo e etsang gradient joalo ka sucrose kapa iodixanol. Mokhoa ona o ka sebelisoa ka tsela ena:

Mohato oa 1: Ho Lokisetsa Gradient ea Botenya

Sesebediswa sa ho lekanya boholo ba di-gradient, jwalo ka sucrose, se lokiswa ka hara tube ya centrifuge, ka bongata bo fapaneng ho tloha hodimo ho ya tlase kapa tlase ho ya hodimo. Sebopeho le boima ba sesebediswa sa ho lekanya boholo ba di-gradient di kgethwa ka hloko, ho netefatsa hore se bobebe ho feta di-peroxisome empa se teteaneng ho feta di-lysosome.

Mohato oa 2: Ho Beha Sampole ka Mealo

Karolo ea lysosome e tsosolositsoeng le peroxisome e fumanoeng ho tsoa ho differential centrifugation e beoa ka hloko holim'a density gradient medium.

Mohato oa 3: Ho kenya centrifugation

Ka mora moo, phaephe e kenngwa ka lebelo le phahameng la centrifugation (ultracentrifugation) ho arola lysosomes le peroxisomes ho latela bongata ba tsona ba ho phaphamala. Nakong ya centrifugation, di-organelles di tsamaya ka hara sebaka seo ho sona bongata ba tsona ba ho phaphamala bo tsamaellanang le boima ba sebaka se potolohileng, di etsa meqhaka e fapaneng.

Tlhahlobo le Litšebeliso tse ling

Kamora karohano e atlehileng, likaroloana tsa lysosome le peroxisome tse fumanoeng li ka hlahlojoa haholoanyane ho sebelisoa mekhoa e fapaneng ho lekola bohloeki ba tsona, boholo, sebopeho le thepa ea ts'ebetso. Mekhoa e kang microscopy ea electron, immunocytochemistry, le liteko tsa ts'ebetso ea enzyme li ka fana ka temohisiso ea bohlokoa mabapi le likaroloana tse arohaneng, e leng se tiisang boitsebahatso ba tsona le bohloeki ba tsona.

Ka ntle ho merero ea lipatlisiso, bokhoni ba ho arola lysosomes le peroxisomes ka ho sebelisa centrifugation bo na le lits'ebetso ho hlahlobisiseng bongaka, nts'etsopele ea lithethefatsi le mekhoa ea phekolo. Ka ho utloisisa mesebetsi ea li-organelles tsena hamolemo, bo-rasaense ba ka hlahloba maano a macha a phekolo a shebaneng le mathata a lysosomes kapa peroxisomal, a kang mafu a polokelo ea lysosomes kapa mathata a peroxisome biogenesis.

Qetellong, karohano ea li-lysosome ho li-peroxisome ka ho sebelisa mekhoa ea centrifugation, joalo ka differential centrifugation le density gradient centrifugation, ke mokhoa oa bohlokoa lipatlisisong tsa baeloji ea sele. Mekhoa ena e nolofalletsa bo-rasaense ho fumana likaroloana tse hloekisitsoeng tsa li-lysosome le li-peroxisome, e leng se thusang ho utloisisa mesebetsi ea tsona ka bomong, litsela tsa metabolism, le mekhoa e ka bang teng ea phekolo bakeng sa mafu a amanang le eona.