Eriksson Emma, Engström David, Scrimgeour Jan, Goksör Mattias
Department of Physics, University of Gothenburg, Gothenburg, Sweden.
Opt Express. 2009 Mar 30;17(7):5585-94. doi: 10.1364/oe.17.005585.
Experiments on single cells are currently gaining more and more interest. Single cell studies often concerns the spatio-temporal distribution of fluorescent proteins inside living cells, visualized using fluorescence microscopy. In order to extract quantitative information from such experiments it is necessary to image the sample with high spatial and temporal resolution while keeping the photobleaching to a minimum. The analysis of the spatial distribution of proteins often requires stacks of images at each time point, which exposes the sample to unnecessary amounts of excitation light. In this paper we show how holographic optical tweezers combined with image analysis can be used to optimize the axial position of trapped cells in an array in order to bring the nuclei into a single imaging plane, thus eliminating the need for stacks of images and consequently reducing photobleaching. This allows more images to be collected, as well as increasing the time span and/or the time resolution in time lapse studies of single cells.
目前，针对单细胞的实验越来越受到关注。单细胞研究通常涉及活细胞内荧光蛋白的时空分布，通过荧光显微镜进行可视化观察。为了从此类实验中提取定量信息，有必要在将光漂白降至最低的同时，以高空间和时间分辨率对样本进行成像。蛋白质空间分布的分析通常需要在每个时间点获取图像堆栈，这会使样本暴露于不必要的激发光量下。在本文中，我们展示了如何将全息光镊与图像分析相结合，用于优化阵列中捕获细胞的轴向位置，以便将细胞核置于单个成像平面中，从而无需图像堆栈，进而减少光漂白。这使得能够收集更多图像，同时增加了单细胞延时研究中的时间跨度和/或时间分辨率。
