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Brownian motion of polyphosphate complexes in yeast vacuoles: characterization by fluorescence microscopy with image analysis

机译：酵母液泡中多磷酸盐复合物的布朗运动：通过荧光显微镜和图像分析进行表征

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摘要

In the vacuoles of Saccharomyces cerevisiae yeast cells, vividly moving insoluble polyphosphate complexes (IPCs) <1 mu m size, stainable by a fluorescent dye, 4',6-diamidino-2-phenylindole (DAPI), may appear under some growth conditions. The aim of this study was to quantitatively characterize the movement of the IPCs and to evaluate the viscosity in the vacuoles using the obtained data. Studies were conducted on S. cerevisiae cells stained by DAPI and fluorescein isothyocyanate-labelled latex microspheres, using fluorescence microscopy combined with computer image analysis (ImageJ software, NIH, USA). IPC movement was photorecorded and shown to be Brownian motion. On latex microspheres, a methodology was developed for measuring a fluorescing particle's two-dimensional (2D) displacements and its size. In four yeast cells, the 2D displacements and sizes of the IPCs were evaluated. Apparent viscosity values in the vacuoles of the cells, computed by the Einstein-Smoluchowski equation using the obtained data, were found to be 2.16 +/- 0.60, 2.52 +/- 0.63, 3.32 +/- 0.9 and 11.3 +/- 1.7 cP. The first three viscosity values correspond to 30-40% glycerol solutions. The viscosity value of 11.3 +/- 1.7 cP was supposed to be an overestimation, caused by the peculiarities of the vacuole structure and/or volume in this particular cell. This conclusion was supported by the particular quality of the Brownian motion trajectories set in this cell as compared to the other three cells. Copyright
机译：在酿酒酵母酵母细胞的液泡中，在某些生长条件下可能会出现小于1微米大小的生动移动的不溶性多磷酸盐复合物（IPC），可被荧光染料4'，6-二mid基-2-苯基吲哚（DAPI）染色。这项研究的目的是定量表征IPC的运动，并使用获得的数据评估液泡中的粘度。使用荧光显微镜结合计算机图像分析（ImageJ软件，NIH，美国），对用DAPI和荧光素异硫氰酸酯标记的乳胶微球染色的酿酒酵母细胞进行了研究。 IPC运动被记录下来并显示为布朗运动。在乳胶微球上，开发了一种用于测量荧光粒子的二维（2D）位移及其大小的方法。在四个酵母细胞中，评估了IPC的二维位移和大小。使用获得的数据通过爱因斯坦-斯莫卢霍夫斯基方程计算的细胞液泡中的表观粘度值为2.16 +/- 0.60、2.52 +/- 0.63、3.32 +/- 0.9和11.3 +/- 1.7 cP 。前三个粘度值对应于30-40％的甘油溶液。 11.3 +/- 1.7 cP的粘度值被认为是高估了，这是由该特定细胞中液泡结构和/或体积的特殊性引起的。与其他三个单元格相比，此单元格中设置的布朗运动轨迹的特殊质量支持了该结论。版权

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