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慶大霉素(GEN)試劑盒
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更新時(shí)間:2024-12-09 21:00:07

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SIGMA現(xiàn)貨,SIGMA代理

慶大霉素(GEN)試劑盒

慶大霉素(GEN)試劑盒報(bào)價(jià),咨詢選購(gòu),技術(shù)服務(wù),選擇正確的實(shí)驗(yàn)試劑或材料是實(shí)驗(yàn)成功的關(guān)鍵。

  一、客戶收到細(xì)胞先不開蓋,放在培養(yǎng)箱靜置若干小時(shí)后(看細(xì)胞密度而定)在倒置顯微鏡下觀察細(xì)胞生長(zhǎng)情況,并對(duì)細(xì)胞進(jìn)行不同倍數(shù)拍照(建議受收細(xì)胞時(shí)的培養(yǎng)基拍一張照片,觀察培養(yǎng)基的顏色和是否有漏液情況,顯微鏡下拍細(xì)胞100X,200X各一張),排除細(xì)胞本身污染的情況;

  收到細(xì)胞未開封,出現(xiàn)污染狀況我們負(fù)責(zé)免費(fèi)發(fā)送一株細(xì)胞。

  二、如為貼壁細(xì)胞,請(qǐng)?jiān)陲@微鏡下觀察細(xì)胞密度:

  1. 未超過(guò)80%匯合度時(shí),用75%酒精(建議配置75%酒精的水也是滅菌超純水,噴灑整個(gè)瓶消毒后放到超菌臺(tái)內(nèi),嚴(yán)格無(wú)菌操作;然后打開蓋子,先把培養(yǎng)基吸出10ml左右,放在離心管里,然后瓶口過(guò)火(嚴(yán)禁直接傾倒),這樣可以保證不污染,再用10ml的移液管,將剩余的培液全部吸出,放于離心管中,培養(yǎng)瓶中只剩余10ml左右培液就可以了。培養(yǎng)16hr(時(shí)間根據(jù)細(xì)胞生長(zhǎng)情況調(diào)整)之后,傳代或者凍存。

  2. 超過(guò)80%匯合度時(shí),請(qǐng)按細(xì)胞培養(yǎng)條件傳代培養(yǎng)。具體步驟如下:

  1) 棄去培養(yǎng)液,用3ml PBS(不含鈣,鎂離子)洗1-2次;

  2) 加1ml消化液(0.25%Trypsin-0.53mM EDTA)于T25培養(yǎng)瓶中,倒轉(zhuǎn)放于37度培養(yǎng)箱1-3分鐘預(yù)熱,然后又將培養(yǎng)瓶倒轉(zhuǎn)大約30秒后,在倒置顯微鏡下觀察細(xì)胞消化情況,若細(xì)胞大部分變圓分散,輕敲幾下培養(yǎng)培養(yǎng)瓶,細(xì)胞隨即脫落下來(lái);

  3) 加入6-8ml*培養(yǎng)基,吸出,分到新的培養(yǎng)瓶中,按要求分瓶。

  三、如為懸浮細(xì)胞,吸出培養(yǎng)液,1000轉(zhuǎn)/分鐘離心5分鐘,吸出上清,管底細(xì)胞用新鮮培養(yǎng)基懸浮細(xì)胞后移回培養(yǎng)瓶。

  注意:換液時(shí)一半用我們的培養(yǎng)基,一半用你們的,以免細(xì)胞不適應(yīng)而造成生長(zhǎng)不好。

  四、細(xì)胞出現(xiàn)問(wèn)題,可重發(fā)的情況有哪些?判定標(biāo)準(zhǔn)是什么?

  (1)細(xì)胞運(yùn)輸途中遭遇的各種問(wèn)題,細(xì)胞丟失、破損、漏液等,重發(fā);

  (2)凍存的細(xì)胞復(fù)蘇后,絕大多數(shù)細(xì)胞未存活,重發(fā);

  (3)存活的細(xì)胞靜置24小時(shí)后,絕大多數(shù)細(xì)胞未存活,重發(fā);

  (4)凍存的細(xì)胞復(fù)蘇后或存活的細(xì)胞靜置4小時(shí)后并且未開封,出現(xiàn)污染,重發(fā);

  (5)視具體情況而定。 

    細(xì)胞培養(yǎng)技術(shù)3個(gè)原則:1、用自己的一套試劑,不要和別人混用;2、勤觀察細(xì)胞,像養(yǎng)寵物一樣喜愛(ài)和關(guān)心;3、有規(guī)律地?fù)Q液和傳代,不要“三天打魚,兩天曬網(wǎng)”。凍存細(xì)胞時(shí)間為5-7個(gè)工作日,活細(xì)胞為7-15個(gè)工作日。

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