Tips na asali don murfin kwayar halitta

Tips na asali don murfin kwayar halitta

Kwayoyin suna ci gaba da cigaba da ayyukan haɓakawa na al'ada, waɗanda ke buƙatar sa hannu na daban-daban. Lokacin da zazzabi ya sauka ƙasa -70 ° C, kariya ta daina aiki. Sabili da haka, a cikin yanayi tare da ƙananan ƙananan yanayin zafi, sel na iya dakatar da ayyukansu na yau da kullun kuma shigar da wani yanayin da ke ba da damar dogon lokaci.

1. Abubuwan gwaji

Kayan kayan gini:

Matsakaici na matsakaici na al'ada

Magani (na al'ada fbs / nbs)

Ulture Workbench

Bakararre Dimethyl sulfixide (Dmso)

Bakararre pbs phosphate buffer gishiri

Bindiga ban ruwa

Bindiga butet ɗin lantarki

Shiri na gwaje-gwajen

A) Shirye-shiryen maganin daskarewa:

Janar Sells: 55% Matsakaicin Matsakaici + 40% na Serum (FBs / Nbs) + 5% DMSo

Sel mai mahimmanci: 90% gyaran magani (FBs / nbs) + 10% DMSO

Amaquot da aka gina karin bayani game da bayani a cikin bututu na 15 ml [gida] da adana a 4 ° C don amfani daga baya.

(B) Shirye-shiryen sel don su daskare:

Kafin daskarewa sel, zaɓi sel tare da kyakkyawan yanayi mai kyau kuma a cikin ci gaban logarithmic na ci gaba kuma a maye gurbinsu da sa'o'i na 12-24 kafin daskarewa don kula da yanayin tantanin halitta.

2. Hanyar gwaji

1. Lura da yawaitar sel don zama daskararre, wanda kusan 80% ~ 90%. Yi amfani da bindiga mai ban sha'awa don ɗaukar tsoffin matsakaici, ƙara broile bakararre don wanke sel sau 1, kuma cire sauran matsakaiciyar yanayin al'adun.

2. Theara adadin da ya dace na trypsin ko ruwan dare na narkewa don ba da damar trypsin shigar da sel, kuma sanya su a cikin incubator don narkewar narkewa. Lura da yanayin sel a karkashin microscope: Cytoplasm yana komawa da sel ba a haɗa shi cikin zanen gado. A wannan gaba, ƙara tasha don dakatar da tsarin narkewa.

3. A hankali ya ɗauki sel tare da tip don ƙirƙirar dakatarwar sel da centrifue the Sell Doge a 1000 Rpm na 3-5 minti.

Jefar da Superatat, ƙara adadin daskarewa na daskarewa, kuma a hankali pipette don sanya sel crounty county. Daidaita yawan ƙwayoyin cuta tare da matsakaici mai kukan don yin sauye-sauye na ƙarshe 5 × 106 / ml ~ 1 × 107 / ml.

4. Yi amfani da bindiga na pipette don ware bututu na Cryobenic bisa ƙarfin damar da ake tsammanin, kuma yi amfani da injin cajin atomatik zuwa caje da hatimi.

5. Tsarin shirin Cryopreatide na sanyi ne na -1 ° C zuwa -2 ° C / min, da sel don zama mai sanyi a gwargwadon matakan: zazzabi dakin → 4 ° C (20min) → - 20 ° C (30min) → -80 ° C ° C (na dare) → dogon lokaci ajiya a cikin ruwa nitrogen.

Hakanan za'a iya adana kai tsaye a cikin injin daskarewa a -80 ° C na dare sannan ya canza zuwa ruwa nitrogen don ajiya.

Lafiya ta Yongyue ita ce babbar masana'antar fasaha ta R & D, samarwa da kuma tallace-tallace masu amfani da kwayoyin halitta. Pipette tukwici, butettes, farantin komputa, samfuran al'adantaccen kayan gwaje-gwaje da sauran wuraren binciken halitta na halitta. Yangyue Mejiniya ya cancanci dogaro da amincinka! Barka da neman shawara!