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利用双光子激发荧光显微镜绘制红细胞鬼影中的血红蛋白残留
Mapping of hemoglobin residuals in erythrocyte ghosts using two photon excited fluorescence microscopy |
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| 课程网址: |
http://videolectures.net/biophysics2018_krmpot_fluorescence_micro...
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| 主讲教师: |
Aleksandar Krmpot
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| 开课单位: |
贝尔格莱德大学
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| 开课时间: |
2018-07-09
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| 课程语种: |
英语
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| 中文简介: |
我们利用双光子激发荧光（TPEF）显微镜来绘制残余血红蛋白的空间分布图。自制的NLSM实验装置利用了730nm钛宝石激光器的飞秒脉冲序列，重复频率76MHz，脉冲持续时间160fs。猪屠宰场血液和人类过期血液被用作起始生物材料。红细胞鬼影是通过逐渐低张溶血制备的[3]。首先，我们使用TPEF显微镜在单细胞水平上对完整的红细胞进行成像，并研究其形态、人的盘状细胞和猪红细胞的棘突细胞。我们已经证明，血红蛋白在完整红细胞中的分布遵循细胞的形状，在细胞膜附近具有明显的丰度[4]。TPEF图像还显示，尽管在逐渐低渗溶血后进行了广泛的冲洗程序，残余血红蛋白仍局限于鬼影膜的细胞内侧[4]。TPEF估计的完整红细胞中的血红蛋白分布和红细胞鬼影中的残余血红蛋白分布在生物技术过程中具有重要意义，但在不同病理条件的诊断中也具有重要意义。
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| 课程简介: |
We have utilized the two photon excitation fluorescence (TPEF) microscopy to map the spatial distribution of residual hemoglobin. The home-made experimental set up for NLSM utilizes the train of the femtosecond pulses from Ti:Sapphire laser at 730nm, repetition rate 76MHz, and pulse duration 160fs. Porcine slaughterhouse blood and human outdated blood were used as a starting biological material. The erythrocyte ghosts were prepared by gradual hypotonic hemolysis [3]. First we used TPEF microscopy to image the intact erythrocytes at single cell level and to study their morphologies, discocyte of human and echynocite of porcine erythrocytes. We have shown that the distribution of hemoglobin in intact erythrocytes follows the cells’ shape with pronounced abundance in the proximity of cell membrane [4]. The TPEF images have also revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, residual hemoglobin localized on intracellular side of the ghost membranes [4]. The TPEF estimated hemoglobin distribution in intact erythrocytes and residual hemoglobin distribution in erythrocyte ghosts could be of importance in biotechnological processes but also in diagnosis of different pathological conditions.
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| 关 键 词: |
双光子; 绘制残余血红蛋白; 荧光显微镜
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| 课程来源: |
视频讲座网
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| 数据采集: |
2021-11-18：zkj
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| 最后编审: |
2021-11-18：zkj
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| 阅读次数: |
57
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