JP2017074008A - 貪食能評価方法及び蛍光測定方法 - Google Patents
貪食能評価方法及び蛍光測定方法 Download PDFInfo
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Abstract
【解決手段】食細胞の貪食能評価方法であって、全血を含む試料中で、食細胞に蛍光物質を取り込ませ、被測定試料を得るステップと、被測定試料を2分間以上静置するステップと、蛍光物質の蛍光を測定するステップと、測定された蛍光の強度に基づいて、食細胞の貪食能を評価するステップと、を備え、測定するステップは、載置された被測定試料の重力方向の中間領域に励起光を照射し、励起光により蛍光物質で発生した蛍光を検出することを含む、貪食能評価方法。
【選択図】図1
Description
0.2w/v%のpH感受性蛍光粒子(Green E.coli,モレキュラープローブス社製)を含有するリン酸緩衝生理食塩水(PBS)をマイクロチューブに入れ、37℃で保温した。次いで、ランセットで採血した人末梢血3μLを添加し、上下に数回ゆっくりと転倒し混合した。次いで、37℃で2分間静置した後、図1に示す光学系を有する蛍光測定装置にマイクロチューブをセットし、蛍光を測定した。蛍光は10秒程度測定し、その測定値の平均値を算出した(対照蛍光強度)。
0.2w/v%のpH感受性蛍光粒子(Green E.coli)を含有するリン酸緩衝生理食塩水(PBS)をマイクロチューブに入れ、37℃で保温した。次いで、ランセットで採血した人末梢血3μLを添加し、上下に数回ゆっくりと転倒し混合した。次いで、37℃で所定時間(0分間、1分間、2分間、3分間、4分間又は5分間)静置した後、図1に示す光学系を有する蛍光測定装置にマイクロチューブをセットし、蛍光を測定した。蛍光は10秒程度測定し、その測定値の平均値を算出した。
Claims (6)
- 食細胞の貪食能評価方法であって、
全血を含む試料中で、食細胞に蛍光物質を取り込ませ、被測定試料を得るステップと、
被測定試料を2分間以上静置するステップと、
前記蛍光物質の蛍光を測定するステップと、
測定された蛍光の強度に基づいて、食細胞の貪食能を評価するステップと、を備え、
前記測定するステップは、載置された被測定試料の重力方向の中間領域に励起光を照射し、前記励起光により前記蛍光物質で発生した蛍光を検出することを含む、
貪食能評価方法。 - 前記被測定試料を得るステップにおいて、前記蛍光物質の取り込みを120分間以上行う、請求項1に記載の貪食能評価方法。
- 前記静置するステップの前に、前記被測定試料を混和するステップを更に含む、請求項1又は2に記載の貪食能評価方法。
- 前記被測定試料に含まれる前記全血の量が、0.5〜10μLである、請求項1〜3のいずれか一項に記載の貪食能評価方法。
- 全血を含む試料を使用して、細胞に取り込まれた蛍光物質の蛍光を測定する蛍光測定方法であって、
全血を含む試料中で、細胞に蛍光物質を取り込ませ、被測定試料を得るステップと、
被測定試料を2分間以上静置するステップと、
前記蛍光物質の蛍光を測定するステップと、を備え、
前記測定するステップは、載置された被測定試料の重力方向の中間領域に励起光を照射し、前記励起光により前記蛍光物質で発生した蛍光を検出することを含む、
蛍光測定方法。 - 前記静置するステップの前に、前記被測定試料を混和するステップを更に含む、請求項5に記載の蛍光測定方法。
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US15/292,453 US10724953B2 (en) | 2015-10-15 | 2016-10-13 | Method for evaluating phagocytic capacity and fluorescence measurement method |
CN201610894378.XA CN106950203B (zh) | 2015-10-15 | 2016-10-13 | 吞噬能力评价方法和荧光测定方法 |
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JP2010508295A (ja) * | 2006-10-27 | 2010-03-18 | ライフ テクノロジーズ コーポレーション | 蛍光発生性pH感受性色素およびその使用方法 |
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CN101846626B (zh) | 2010-05-24 | 2012-07-11 | 天津商业大学 | 一种利用流式细胞仪检测腹腔吞噬细胞活性的方法 |
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SCHREINER, L., ET AL.: "Phagocytosis and digestion of pH-sensitive fluorescent dye (Eos-FP) transfected E. coli in wohle blo", J. CELL COMMUN. SIGNAL., vol. Vol.5, JPN6019010849, 2011, pages 135 - 144 * |
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