Malangizo Oyambira Otetezera Zachikhalidwe cha Cell

Malangizo Oyambira Otetezera Zachikhalidwe cha Cell

Maselo akupitiliza kagayidwe kawo nthawi yofalikira bwino, yomwe imafunikira mapuloteni osiyanasiyana. Kutentha kwatsikira pansipa -70 ° C, mapuloteni amasiya kugwira ntchito. Chifukwa chake, m'malo ocheperako, maselo amatha kuletsa zochitika zawo kagayidwe ndikulowetsa boma lomwe limalola kusungirako kwa nthawi yayitali.

1. Zipangizo zoyesera

Zinthu:

Chikhalidwe Chachikulu

Serramu (FBSALL FBB / NBS)

Ultrapure Corgench

Wosabala Dimethyl Sulfoxide (DMSO)

Ster PBS phosphate Buffer mchere yankho

Mfuti

Mfuti yamagetsi

Kukonzekera kwa kuyesa

A) Kukonzekera yankho la kuzizira:

Maselo a General: 55% basal sing'anga + 40% bovine seum (FBS / NBS) + 5% DMSO

Maselo Ofunika: 90% Bovine seum (FBS / NBS) + 10% DMSO

Alliquot yankho lokonzekera mu 15 crentrifuge chubu [chisa] ndi sitolo] 4 ° C pogwiritsa ntchito pambuyo pake.

(b) Kukonzekera maselo kuti mukhale ozizira:

Musanametse maselo, sankhani maselo okula bwino komanso gawo lotakatalika ndikusintha ndi maola 12-24 musanadyetse vuto la cell.

2. Njira yoyesera

1. Onani kachulukidwe ka maselo kuti ukhale wozizira, womwe uli pafupifupi 80% ~ 90%. Gwiritsani ntchito mfuti pipette kuti musiyire sing'anga wakale, onjezerani ma pbs osambitsa maselo 1-2s, ndikuchotsa sing'anga yotsalira pachikhalidwe.

2. Onjezani kuchuluka koyenera kapena madzi amchere kuti alole trypin kuti ilowetse maselo, ndikuyiyika mu chofungatira kuti chimbudzi. Onani mkhalidwe wa maselo omwe ali pansi pa microscope: cytoplasm akulephera ndipo maselo salumikizidwanso m'mapepala. Pakadali pano, onjezani njira yothetsera kugaya.

3. Pang'onopang'ono maselo ndi nsonga kuti mupange kuyimitsidwa kwa cell ndi centrifuge Kuyimitsidwa kwa cell pa 1000 rpm kwa mphindi 3-5.

Tayanitsani sunanatant, onjezani njira yoyenera yothetsera vuto, ndipo chitoliro pang'ono kuti maselowo aziwerengera mofananamo. Sinthani kachulukidwe ka maselo ndi sing'anga yomaliza kuti apange kachulukidwe katatu × 106 / ml ~ 1 × 107 / ml.

4. Gwiritsani ntchito pipette mfuti kuti muletse machubu a christhagenic malinga ndi momwe akuyembekezeredwa, ndikugwiritsa ntchito makina opangira zokha ku chipewa ndi chisindikizo.

5. Gawo lokhazikika 20 ° C (30min) → -80 ° C ° C (usiku) →

Itha kusungidwanso mwachindunji mu -80 ° C motalika kenako ndikusamukira kumadzi a nayitrogeni kuti isungidwe.

Yongyue Medical ndi bizinesi yapamwamba kwambiri yolowera ku R & D, kupanga ndi kugulitsa zosemphana ndi zinthu zopanda pake. Malangizo a Pipette, ma pipet, mbale ya pcr, zinthu zina zam'manja zopangidwa ndi zinthu zina zothandizira zachilengedwe. Yongyue Medical ndiyofunika kudalirika ndi kusankha kwanu! Takulandilani!