nzira yekuparadzanisa lysosomes kubva muchimiro cheperoxisomes kuburikidza ne centrifugation

Nhanganyaya kuLysosomes nePeroxisomes

Lysosomes neperoxisomes inhengo mbiri dzakasiyana dzemasero dzine mabasa akasiyana akakosha mukushandiswa kwemasero uye homeostasis. Lysosomes inhengo dzakasungwa nemembrane dzakazadzwa nema enzymes ekugaya anokonzera kupwanya marara emasero, zvidimbu zvekunze, uye hutachiona. Kune rumwe rutivi, peroxisomes inobatanidzwa mukuita kwakasiyana-siyana kwemetabolism, kunyanya izvo zvine chekuita nekubvisa chepfu uye kushandiswa kwemafuta mumuviri. Kuti tidzidze idzi nhengo dzakasiyana, imwe nzira inowanzo shandiswa ndeye centrifugation. Muchinyorwa chino, tichaongorora kuti centrifugation ingashandiswa sei kuparadzanisa lysosomes kubva kuperoxisomes zvinobudirira.

Nheyo uye Matekiniki eCentrifugation

Kugadzira centrifugation inzira ine simba inoshandiswa kupatsanura zvikamu zvakasiyana zvemusanganiswa wakasiyana zvichienderana nehukuru hwazvo, huwandu hwazvo, uye chimiro chazvo. Inoshanda pamisimboti yekuumbwa kwevhu, uko zvidimbu zvinorema zvinogara pasi nekukurumidza pasi pesimba re centrifugal. Kuparadzanisa lysosomes kubva ku peroxisomes, nzira dzakasiyana dzekugadzira centrifugation, dzakadai se differential centrifugation uye density gradient centrifugation, dzinogona kushandiswa.

Kusiyanisa Kwekudzora: Kubvisa Lysosomes nePeroxisomes

Kuparadzanisa masero emasero kunosanganisira matanho akasiyana-siyana ekubvisa masero emasero nekumhanya kwakasiyana uye nguva dzakasiyana dzekuparadzanisa ma organelles zvichienderana nehuwandu hwawo. Heino mashandisirwo ainogona kuitwa kubvisa lysosomes ne peroxisomes:

Danho 1: Kubatanidzwa kweHomogenization

Danho rekutanga nderekupwanya masero zvinyoro nyoro woburitsa zviri mukati mawo uchishandisa nzira yekubatanidza masero. Izvi zvinogona kuitwa nekushandisa sonicating kana kushandisa homogenizer yakasarudzika. Musanganiswa unobuda unonzi homogenate.

Danho rechipiri: Kudzora Kumhanya Kwakaderera

Homogenate inobva yaiswa centrifugation ine kumhanya kwakaderera (inenge 1,000-2,000 xg) kwenguva pfupi. Danho iri rinobatsira kuparadzanisa tsvina yakakura yemasero nechikamu chenyukireya kubva kune mamwe ma organelle ari muhomogenate.

Danho rechitatu: Kudzora Kumhanya Kwepakati

Supernatant inowanikwa kubva padanho re centrifugation rinomhanya zvishoma rinoiswa mu centrifugation inomhanya zviri pakati nepakati (inenge 10,000 xg) kuti iparadze organelles dzakakora dzakadai se lysosomes ne peroxisomes. Danho iri rinoguma ne pellet ine organelles idzi.

Danho rechina: Geza uye simbisazve

Pellet yakawanikwa padanho rekare inogezwa nemushonga wakakodzera webuffer uye yozodzorerwa kuti pave nechikamu che lysosome chakacheneswa uye peroxisome.

Kudzora Kuparadzana Kwemagiraidhi Ekuwedzera Density Gradient: Kugadzirisa Kuparadzana

Kushandiswa kwe density gradient centrifugation inzira yepamusoro ye centrifugation inobvumira kucheneswa nekugadziriswa kwe lysosomes ne peroxisomes. Munzira iyi, density gradient inoumbwa uchishandisa gradient-forming solution yakadai se sucrose kana iodixanol. Heino mashandisirwo ayo:

Danho 1: Kugadzirira Density Gradient

Chinhu chinoyera huwandu hwezvinhu zvakaita sesucrose, chinogadzirwa muchubhu ye centrifuge, chine huwandu hwakasiyana kubva kumusoro kusvika pasi kana pasi kusvika kumusoro. Maumbirwo uye huwandu hwechinhu chinoyera huwandu zvinosarudzwa nokungwarira, zvichiita kuti zvive nyore kupfuura peroxisomes asi zvine huremu hwakanyanya kupfuura lysosomes.

Danho rechipiri: Kuisa Muenzaniso muzvikamu

Chikamu che lysosome ne peroxisome chakadzorerwa chinowanikwa kubva mu differential centrifugation chinoiswa pamusoro pe density gradient medium.

Danho rechitatu: Kudzora centrifugation

Chubhu iyi inobva yaiswa centrifugation ine kumhanya kukuru (ultracentrifugation) kuti iparadzanise lysosomes neperoxisomes zvichienderana nekuwanda kwazvo. Munguva ye centrifugation, ma organelles anofamba nepakati kusvika asvika panzvimbo iyo kuwanda kwawo kunoenderana nekuwanda kwenzvimbo yakapoteredza, zvichiita mabhandi akasiyana.

Kuongorora uye Zvimwe Zvishandiso

Mushure mekupatsanurwa zvakanaka, zvikamu zve lysosome ne peroxisome zvinowanikwa zvinogona kuongororwazve uchishandisa nzira dzakasiyana-siyana dzekuongorora kuchena kwazvo, saizi, maumbirwo azvo, uye mashandiro azvo. Matekiniki akadai se electron microscopy, immunocytochemistry, uye enzyme activity assays anogona kupa ruzivo rwakakosha pamusoro pezvikamu zvakaparadzaniswa, zvichisimbisa kuzivikanwa kwazvo uye kuchena kwazvo.

Kunze kwezvinangwa zvekutsvagisa, kugona kuparadzanisa lysosomes neperoxisomes kuburikidza ne centrifugation kune mashandisirwo mukuongorora kwekurapa, kugadzira mishonga, uye kupindira mukurapa. Nekunzwisisa zviri nani mashandiro ezvipenyu izvi, masayendisiti anogona kutsvaga nzira itsva dzekurapa dzinotarisa matambudziko e lysosomes kana peroxisomal, akadai sezvirwere zve lysosomes storage kana peroxisome biogenesis disorders.

Mukupedzisa, kupatsanurwa kwe lysosomes kubva ku peroxisomes uchishandisa nzira dze centrifugation, dzakadai se differential centrifugation uye density gradient centrifugation, inzira inokosha mukutsvagisa kwe biology yemasero. Maitiro aya anogonesa masayendisiti kuwana zvikamu zvakacheneswa zve lysosomes ne peroxisomes, zvichibatsira mukunzwisisa mabasa avo ega ega, nzira dze metabolic, uye nzira dzinogona kushandiswa pakurapa zvirwere zvine chekuita nazvo.