產(chǎn)品詳情
使用方法/ Instructions
前期準(zhǔn)備
1) 將分離液的溫度平衡至室溫（RT），避光。
2) 將分離液用移液管從分離管嵌套的中心孔中加入：15ml的分離管需要加入約4ml分離液；50ml的分離管需要加入約13ml分離液，確保分離液覆蓋嵌套上方。
3) 平衡至室溫后的分離管就可以加入抽取的抗凝血和骨髓了。雖然加入稀釋標(biāo)本的生理鹽水不是十分必要，但是它能幫助提高分離效果。
分離過程
加入細(xì)胞分離液 加入樣本 離心后分層 收集富集細(xì)胞、重復(fù)洗滌
1) 將抗凝樣本（血或骨髓，如果需要可以用生理鹽水稀釋）延管壁緩慢倒入或用移液管延管壁緩慢加入到分離管中：如果用15ml的分離管建議使用4-9ml樣本；如果用50ml的分離管建議使用13-30ml樣本。
2) 室溫下離心，離心力為1200 x g需要10分鐘，關(guān)閉離心機(jī)。對于放置24h以上的樣品，建議離心時間加長。
3) 離心后液體分離情況（從上到下）為：a血漿 ；b富集的細(xì)胞部分（中間相包含淋巴細(xì)胞/PBMCs細(xì)胞）；c分離液；d嵌套；e沉淀（紅細(xì)胞和粒細(xì)胞）。采集或丟棄富集細(xì)胞所在層以上5到10mm的血漿層有助于防止富集細(xì)胞被血小板再次污染。
4) 收獲富集細(xì)胞（淋巴細(xì)胞/PBMCs細(xì)胞），將分離管中上清倒入另一干凈離心管中，分離管中的嵌套能有效避免富集細(xì)胞被紅細(xì)胞和粒細(xì)胞再次污染。建議不要將分離管倒置2s以上。
5) 用磷酸鹽緩沖液（PBS）洗滌富集細(xì)胞（淋巴細(xì)胞/PBMCs細(xì)胞），然后在250 x g離心力下離心10分鐘。
6) 按步驟5重復(fù)洗滌2次，最后用5ml PBS緩沖液重新懸浮細(xì)胞。
注意事項/ Cautions
1) 本產(chǎn)品應(yīng)由經(jīng)過專業(yè)培訓(xùn)的人員操作，并遵守良好的實驗室規(guī)范。
2) 請勿重復(fù)使用分離管。
3) 由于各品牌離心機(jī)的性能不同，地區(qū)溫度環(huán)境差異，可能影響分離效果，用戶可以調(diào)節(jié)離心轉(zhuǎn)速和離心的時間，摸索最佳的分離條件（具體分離條件由各實驗室自定）。
4) 可用于人類外周血、骨髓和臍帶血樣本。它不適用于白細(xì)胞分離樣本、血沉棕黃層樣本或超過48小時的樣本。
5) 離心后，細(xì)胞可能聚集在富集層以上的分離管的管壁上。這種聚合是正常的，受樣本質(zhì)量、樣本放置時間和抗凝劑類型的影響。此聚合與分離管的使用無關(guān)。細(xì)胞可以通過使用移液管尖端刮管壁的一側(cè)來清除。
6) 處理任何生物來源的標(biāo)本，使用采血針、采血管系列、相關(guān)儀器等一定要注意按嚴(yán)格的操作規(guī)程使用。請把標(biāo)本當(dāng)成可能感染 HIV、HBV、HCV等傳染病的危險物質(zhì)處理。為避免操作時感染的危險，請使用一次性手套。
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