Kahn E, Lizard G, Frouin F, Bernengo J C, Souchier C, Bessède G, Clément O, Siitari H, Gambert P, Frija G, Todd-Pokropek A
Institut National de la Santé et de la Recherche Médicale U494, Centre Hospitalier Universitaire Pitié-Salpêtrière, 75634 Paris, France.
Anal Quant Cytol Histol. 2001 Feb;23(1):47-55.
To analyze externalization of phosphatidylserine via annexin V on apoptotic cells by laser scanning confocal microscopy and factor analysis of biomedical image sequences (FAMIS).
Streptavidin-fluorescein isothiocyanate (FITC), -europium (Eu), -phycoerythrin (PE) and -Texas Red (TR) were chosen to reveal the binding of biotinylated annexin V on apoptotic U937 human leukemic cells and ECV-304 human endothelial cells induced under treatment with 7-ketocholesterol or 7 beta-hydroxycholesterol. Excitation of each fluorochrome was obtained by selection of specific lines (351 + 364 nm, 488 nm) of the argon laser of a confocal microscope. Temporal and spectral series were performed to characterize each fluorochrome. FAMIS was applied to these series to estimate images corresponding to stains.
Each fluorochrome was clearly distinguished, and images showed localization of phosphatidylserine, which was improved by image analysis.
On apoptotic cells it is possible to analyze differences in the improved visualization of phosphatidylserine in series processed by FAMIS with the use of biotinylated annexin V revealed with streptavidin-FITC, -Eu, -PE or -TR.
通过激光扫描共聚焦显微镜和生物医学图像序列因子分析（FAMIS）分析凋亡细胞上磷脂酰丝氨酸通过膜联蛋白V的外化情况。
选择链霉亲和素 - 异硫氰酸荧光素（FITC）、 - 铕（Eu）、 - 藻红蛋白（PE）和 - 德克萨斯红（TR）来揭示生物素化膜联蛋白V与经7 - 酮胆固醇或7β - 羟基胆固醇处理诱导的凋亡U937人白血病细胞和ECV - 304人内皮细胞的结合。通过选择共聚焦显微镜氩激光的特定谱线（351 + 364 nm、488 nm）获得每种荧光染料的激发。进行时间和光谱序列以表征每种荧光染料。将FAMIS应用于这些序列以估计与染色相对应的图像。
每种荧光染料都能清晰区分，图像显示了磷脂酰丝氨酸的定位，通过图像分析得到了改善。
对于凋亡细胞，使用链霉亲和素 - FITC、 - Eu、 - PE或 - TR揭示的生物素化膜联蛋白V，通过FAMIS处理的序列可以分析磷脂酰丝氨酸改善可视化方面的差异。
