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Sei tsika dzemasero dzichiisa masero matsvuku eropa kutanga?
2022-12-23
Basic sumo
Erythrocyte lysate ndiyo imwe yenzira dzakareruka uye dziri nyore dzekubvisa masero matsvuku eropa, ndiko kuti, kupatsanura masero matsvuku eropa ne lysate, iyo isingakuvadzi masero ane nucleated uye inogona kubvisa zvizere masero matsvuku eropa. Lysate cleavage inzira yakapfava yekubvisa masero matsvuku eropa, iyo inonyanya kushandiswa kupatsanura uye kucheneswa kwematishu masero akaparadzirwa ne enzyme digestion, kupatsanurwa uye kucheneswa kwemalymphocytes, uye kubviswa kwemasero matsvuku eropa mukuyedzwa kwetishu protein uye nucleic. kubviswa kweasidhi. Masero enyama anowanikwa ne lysate yemasero matsvuku eropa haana masero matsvuku eropa, uye anogona zvakare kushandiswa kune yekutanga tsika, sero fusion, kuyerera cytometry, kupatsanura uye kubviswa kwe nucleic acid neprotein, nezvimwe.
Mirayiridzo yekushandisa
Tissue cell sample
1. Matishu matsva akagayiwa nepancreas / enzyme kana collagenase uye akaparadzirwa mune imwechete sero kumiswa, uye iyo supernatant yakaraswa ne centrifugation.
2. Tora ELS lysate kubva mufiriji pa4â, wedzera ELS lysate kune cell precipitate muchiyero che1: 3-5 (wedzera 3-5ml ye lysate ku 1ml yesero yakasanganiswa), zvinyoronyoro furidzira uye sanganisa.
3. Centrifuge pa 800-1000rpm kwemaminitsi 5-8 uye urasire iyo yepamusoro tsvuku yakajeka mvura.
4. Chikamu chemvura chakaunganidzwa uye centrifuged neHank's solution kana serum-free culture solution ye2-3 nguva.
5, kana kutsemuka kusina kukwana/kukwana kunogona kudzokororwa nhanho 2 ne3.
6. Maseru ekudzoreredza ezviedzo zvinotevera; Kana RNA yakabviswa, zviri nani kuzviita mumhinduro yakagadzirwa kubva kuChikamu 4 uchishandisa DEPC mvura
Erythrocyte lysate ndiyo imwe yenzira dzakareruka uye dziri nyore dzekubvisa masero matsvuku eropa, ndiko kuti, kupatsanura masero matsvuku eropa ne lysate, iyo isingakuvadzi masero ane nucleated uye inogona kubvisa zvizere masero matsvuku eropa. Lysate cleavage inzira yakapfava yekubvisa masero matsvuku eropa, iyo inonyanya kushandiswa kupatsanura uye kucheneswa kwematishu masero akaparadzirwa ne enzyme digestion, kupatsanurwa uye kucheneswa kwemalymphocytes, uye kubviswa kwemasero matsvuku eropa mukuyedzwa kwetishu protein uye nucleic. kubviswa kweasidhi. Masero enyama anowanikwa ne lysate yemasero matsvuku eropa haana masero matsvuku eropa, uye anogona zvakare kushandiswa kune yekutanga tsika, sero fusion, kuyerera cytometry, kupatsanura uye kubviswa kwe nucleic acid neprotein, nezvimwe.
Mirayiridzo yekushandisa
Tissue cell sample
1. Matishu matsva akagayiwa nepancreas / enzyme kana collagenase uye akaparadzirwa mune imwechete sero kumiswa, uye iyo supernatant yakaraswa ne centrifugation.
2. Tora ELS lysate kubva mufiriji pa4â, wedzera ELS lysate kune cell precipitate muchiyero che1: 3-5 (wedzera 3-5ml ye lysate ku 1ml yesero yakasanganiswa), zvinyoronyoro furidzira uye sanganisa.
3. Centrifuge pa 800-1000rpm kwemaminitsi 5-8 uye urasire iyo yepamusoro tsvuku yakajeka mvura.
4. Chikamu chemvura chakaunganidzwa uye centrifuged neHank's solution kana serum-free culture solution ye2-3 nguva.
5, kana kutsemuka kusina kukwana/kukwana kunogona kudzokororwa nhanho 2 ne3.
6. Maseru ekudzoreredza ezviedzo zvinotevera; Kana RNA yakabviswa, zviri nani kuzviita mumhinduro yakagadzirwa kubva kuChikamu 4 uchishandisa DEPC mvura
Masero matsvuku eropa ane hupenyu hupfupi kwazvo, mazuva 120 chete, asi anobereka ropa nekukurumidza, uye muchiitiko ichi ivo vanonyanya kukwanisa kupatsanura masero, uye ndivo vanokurumidza kupatsanura masero evose, saka sero iri rinokosha zvikuru. saka inobatsira zvikuru kune tsika yesero. Zviri nyore kwazvo, haina chero organelles mairi, chete cell membranes uye mapuroteni.
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