Tokumasu F, Dvorak J
Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0425, USA.
J Microsc. 2003 Sep;211(Pt 3):256-61. doi: 10.1046/j.1365-2818.2003.01219.x.
Recent developments in quantum dot technology have resulted in the introduction of new fluorescence immunocytochemical probes. In contrast to organic fluorophores, which are not photostable, the high quantum yield and remarkable photostability of quantum dots solve major problems associated with immunocytochemical studies of erythrocytes. We report here the first application of quantum dots to immunocytochemical studies of human erythrocytes capable of being used in high-magnification, three-dimensional erythrocyte reconstruction techniques. The procedure consists of stabilizing human erythrocytes with a homofunctional imidoester cross-linker to minimize fixative-induced autofluorescence followed by reacting with a quantum dot - monoclonal antibody complex to label band 3 protein. Our new procedure clearly showed a non-homogeneous, raft-like distribution of band 3 protein in the erythrocyte membrane. We also demonstrate the applicability of our technique to studies of erythrocyte membrane modifications occurring during the invasion of a malaria parasite.
量子点技术的最新进展带来了新型荧光免疫细胞化学探针。与不耐光的有机荧光团不同，量子点的高量子产率和出色的光稳定性解决了与红细胞免疫细胞化学研究相关的主要问题。我们在此报告量子点在人类红细胞免疫细胞化学研究中的首次应用，该研究可用于高倍放大的三维红细胞重建技术。该过程包括用同功能亚胺酯交联剂稳定人类红细胞，以尽量减少固定剂诱导的自发荧光，然后与量子点 - 单克隆抗体复合物反应以标记带3蛋白。我们的新方法清楚地显示了带3蛋白在红细胞膜中呈非均匀的筏状分布。我们还证明了我们的技术在研究疟原虫入侵期间发生的红细胞膜修饰方面的适用性。
