Imaging and Cytometry Laboratory, Bioscience Technology Facility, Department of Biology, University of York, UK.
Sci Rep. 2013;3:2369. doi: 10.1038/srep02369.
Cell imaging often relies on synthetic or genetic fluorescent labels, to provide contrast which can be far from ideal for imaging cells in their in vivo state. We report on the biological application of a, label-free, high contrast microscopy technique known as ptychography, in which the image producing step is transferred from the microscope lens to a high-speed phase retrieval algorithm. We demonstrate that this technology is appropriate for label-free imaging of adherent cells and is particularly suitable for reporting cellular changes such as mitosis, apoptosis and cell differentiation. The high contrast, artefact-free, focus-free information rich images allow dividing cells to be distinguished from non-dividing cells by a greater than two-fold increase in cell contrast, and we demonstrate this technique is suitable for downstream automated cell segmentation and analysis.
细胞成像是通过合成或遗传荧光标记来实现的,然而,这些标记提供的对比度远非理想,无法满足对活细胞进行成像的要求。我们报告了一种免标记、高对比度显微镜技术的生物学应用,该技术称为相衬术,其成像步骤从显微镜透镜转移到高速相位恢复算法。我们证明了该技术适用于贴壁细胞的免标记成像,特别适合报告细胞分裂、细胞凋亡和细胞分化等细胞变化。高对比度、无伪影、无焦点、信息丰富的图像使得可以通过细胞对比度增加两倍以上来区分有丝分裂和非有丝分裂的细胞,并且我们证明该技术适用于下游的自动细胞分割和分析。