Huazhong University of Science and Technology, Wuhan National Laboratory for Optoelectronics, Britton Chance Center for Biomedical Photonics, Wuhan, Hubei, China.
J Biomed Opt. 2010 Jul-Aug;15(4):045003. doi: 10.1117/1.3465595.
The optical properties of antiphotobleaching and the advantage of long-term fluorescence observation of quantum dots are fully adopted to study the effects of iron on the endocytosis of transferrin. Quantum dots are labeled for transferrin and endocytosis of transferrin in HeLa cells is observed under the normal state, iron overloading, and an iron-deficient state. In these three states, the fluorescence undergoes a gradual process of first dark, then light, and finally dark, indicating the endocytosis of transferrin. The fluorescence intensity analysis shows that a platform emerges when fluorescence changes to a certain degree in the three states. Experienced a same period of time after platform, the fluorescence strength of cells in the normal state is 1.2 times the first value, and the iron-deficiency state is 1.4 times, but the iron overloading state was 0.85 times. We also find that the average fluorescence intensity in cells detected by the spectrophotometer in the iron-deficiency state is almost 7 times than that in a high iron state. All this proves that iron overloading would slow the process, but iron deficiency would accelerate endocytosis. We advance a direct observational method that may contribute to further study of the relationship of iron and transferrin.
充分利用量子点耐光漂白和长期荧光观察的优点,研究铁对转铁蛋白内吞作用的影响。将量子点标记为转铁蛋白,并在正常状态、铁过载和缺铁状态下观察 HeLa 细胞中的转铁蛋白内吞作用。在这三种状态下,荧光经历了一个逐渐的过程,先是变暗,然后变亮,最后再次变暗,表明转铁蛋白发生了内吞作用。荧光强度分析表明,在三种状态下,荧光变化到一定程度时会出现一个平台。在平台出现后的相同时间段内,正常状态下细胞的荧光强度是初始值的 1.2 倍,缺铁状态下是 1.4 倍,但铁过载状态下仅为 0.85 倍。我们还发现,分光光度计检测到缺铁状态下细胞的平均荧光强度几乎是高铁状态下的 7 倍。所有这些都证明铁过载会减缓这一过程,但缺铁会加速内吞作用。我们提出了一种直接观察的方法,可能有助于进一步研究铁与转铁蛋白的关系。