利用图像处理技术提高 SNARF-1 染色植物细胞的细胞质 pH 值测量精度。
Improved cytoplasmic pH measurements in SNARF-1 stained plant cells by image processing.
机构信息
Faculty of Bioscience and Applied Chemistry, Hosei University, Koganei, Tokyo, Japan.
出版信息
Plant Signal Behav. 2010 Apr;5(4):406-8. doi: 10.4161/psb.5.4.10804. Epub 2010 Apr 30.
Cytoplasmic pH has long been considered to act as a secondary messenger of various cellular responses by affecting the ionization state of proteins. In plant biology, cytoplasmic pH has traditionally been measured, especially in guard cells and as a response to plant microorganism interactions, with pH-sensitive microelectrodes. More recently, the development of fluorescent pH markers, such as BCECF and SNARF-1, has allowed us to monitor cytoplasmic pH without the need for electrophysiological equipment. However, because of vacuolar structures that occupy a large volume of plant cells, simple measurements of fluorescent intensities are insufficient to provide precise cytoplasmic pH values. In this addendum, we describe our improved method to monitor cytoplasmic pH in plant cells stained by SNARF-1 by image processing using a noise-reducing filter after determination of an optimal ROI size. In addition, further developments for automated region extraction are proposed.
细胞质 pH 一直被认为是通过影响蛋白质的离解状态来充当各种细胞反应的第二信使。在植物生物学中，细胞质 pH 值一直以来都是通过 pH 敏感型微电极来测量的，尤其是在保卫细胞中，以及作为对植物与微生物相互作用的响应。最近，荧光 pH 标记物（如 BCECF 和 SNARF-1）的发展使我们能够在不需要电生理设备的情况下监测细胞质 pH 值。然而，由于液泡结构占据了植物细胞的大部分体积，因此简单地测量荧光强度不足以提供精确的细胞质 pH 值。在本增补中，我们描述了一种改进的方法，通过在确定最佳 ROI 大小后使用降噪滤波器对 SNARF-1 染色的植物细胞进行图像处理，从而监测细胞质 pH 值。此外，还提出了用于自动区域提取的进一步发展。
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