Microscopy raw and processed data to measure cell size, cell death and multi-nucleated ratio in yeast cells.

The data contains the raw data folder and a processed folder. The raw data is divided into Bright Field experiments and DAPI. The BrightField Microscopy images were taken on the strains bem1::KanMx pGal-Cdc42-sfGFP,bem1bem3::CloNAT pGal-Cdc42-sfGFP,pGal-Cdc42-sfGFP, and WT. The goal was to measure the cell sizes as a proxy of the polarization state for those strains in specific galactose concentrations taken by the population growth results.  We use three galactose concentrations : 0%, 0.06%, and 0.1%. %=w/vThe processed folder contains an Excel file with three sheets: cell radii ratio, cell death radio,, and multinucleated cells ratio with the post-processed relevant variables from the raw data. All microscopy was performed with a Nikon Eclipse Ti-E inverted microscope with an oil immersion 60x objective,1.40 of numerical aperture, and a refractive index of 1.51. The software used for data collection was Nis-Elements Ar (https://www.microscope.healthcare.nikon.com/products/software/nis-elements/nis-elements-advanced-research) version 4.51.<br>Cells that do not polarize are predicted to have nuclear divisions but no cellular division, and therefore, we hypothesize that cells with polarization defects are more likely to be multinucleate. We measured the percentage of multinucleated cells for the different genetic backgrounds and galactose concentrations using DAPI staining combined with fluorescence microscopy. The DAPI staining protocol was taken from doi: dx.doi.org/10.17504/protocols.io.eigbcbw. Samples were imaged in a cover slip right after the DAPI staining. We used a Nikon Eclipse Ti-E inverted microscope with an oil immersion 60x objective and 1.40 numerical aperture. Its refractive index is 1.51. We imaged in two channels, namely, Brightfield and DAPI Fluorescence. The emission wavelength was 450 nm, and the excitation wavelength was 395 nm. The exposure time was 70ms, and the laser power was 1\%. The software used for data collection was Nis-Elements Ar (https://www.microscope.healthcare.nikon.com/products/software/nis-elements/nis-elements-advanced-research) version 4.51. Data analysis was performed with Cell Counter[https://imagej.nih.gov/ij/plugins/cell-counter.html ] plugin from ImageJ, version 1.53t, which requires Java 1.8.0_172(64-bit). Each cell was labeled to count the total number of cells in each frame. Further, each cell with two or more nuclei and the dead cells were differently labeled.

本数据集包含原始数据文件夹与预处理后数据文件夹。原始数据分为明场（Bright Field）实验与DAPI染色数据两类。明场显微成像的样本对应如下菌株：bem1::KanMx pGal-Cdc42-sfGFP、bem1bem3::CloNAT pGal-Cdc42-sfGFP、pGal-Cdc42-sfGFP以及野生型（WT）。本研究的目标为，通过测量不同半乳糖浓度下各菌株的细胞尺寸，以其作为细胞极化状态的替代指标，该浓度梯度基于群体生长实验结果设定。本次实验采用的半乳糖浓度共3种：0%、0.06%与0.1%（质量体积比，w/v）。
预处理文件夹内含1个Excel文件，包含3个工作表：细胞半径比值表、细胞死亡率比值表与多核细胞占比表，汇总了从原始数据中提取的经后处理的相关变量。所有显微成像实验均采用尼康（Nikon）Eclipse Ti-E倒置显微镜，配置60倍油浸物镜（数值孔径1.40，浸液折射率1.51）；数据采集软件为Nis-Elements Ar v4.51，官方链接：https://www.microscope.healthcare.nikon.com/products/software/nis-elements/nis-elements-advanced-research。
研究推测，无法完成极化的细胞仅会发生核分裂而不会进行胞质分裂，因此我们提出假设：存在极化缺陷的细胞更易出现多核表型。我们结合DAPI染色与荧光显微成像，统计了不同遗传背景菌株在各半乳糖浓度下的多核细胞占比。DAPI染色流程引自文献：dx.doi.org/10.17504/protocols.io.eigbcbw。DAPI染色完成后立即在盖玻片上对样本进行成像，成像仍采用前述尼康Eclipse Ti-E倒置显微镜，配置60倍油浸物镜（数值孔径1.40，浸液折射率1.51）。本次成像采用双通道模式：明场通道与DAPI荧光通道。DAPI荧光通道的激发波长为395 nm，发射波长为450 nm；曝光时长70 ms，激光功率1%。数据采集软件为Nis-Elements Ar v4.51，官方链接：https://www.microscope.healthcare.nikon.com/products/software/nis-elements/nis-elements-advanced-research。数据分析采用ImageJ v1.53t的Cell Counter插件（官方链接：https://imagej.nih.gov/ij/plugins/cell-counter.html），该插件依赖Java 1.8.0_172（64位版本）。实验中对每帧图像内的所有细胞进行标注以统计总细胞数；此外，对含有2个及以上细胞核的细胞与死亡细胞分别采用不同的标注方式。