Data underlying Chapter 4 of PhD thesis: Working toward establishing the link between foci, filaments and membrane abundance

This collection of microscopy datasets investigates how the essential phospholipid synthesis enzyme PlsB regulates membrane growth and organization in <em>Escherichia coli</em>. Fluorescence microscopy data capture PlsB-msGFP2 localization during spheroplast formation, osmotic shocks, FRAP experiments, and altered protein expression levels. These experiments reveal how PlsB dynamics change when fatty-acid synthesis is inhibited, the cell wall is weakened, or membrane area is perturbed. Additional widefield fluorescence imaging of PlsB point-mutant strains assesses how individual amino acid substitutions affect protein clustering and membrane association. Electron microscopy datasets complement these observations by examining ultrastructural context in minicells, thin-sectioned cells, and purified PlsB samples under different preparation conditions. Negative stain and cryo-TEM imaging allow evaluation of potential PlsB oligomerization and membrane interaction at high resolution. Together, the fluorescence and EM datasets provide multiscale insight into the spatial organization of PlsB and its coupling to membrane biogenesis. Furthermore, images are provided showing successful formation of vesicles which can in the future be used as vehicles for in vitro study of PlsB. A MATLAB analysis script supports quantitative extraction of cell geometry and PlsB foci statistics, enabling reproducible interpretation of these imaging data.

本套显微镜数据集旨在探究磷脂合成必需酶PlsB如何调控大肠杆菌（Escherichia coli）的膜生长与膜组织模式。荧光显微镜数据记录了原生质球形成、渗透压冲击、荧光漂白恢复（FRAP）实验以及蛋白表达水平改变过程中PlsB-msGFP2的定位情况。上述实验揭示了脂肪酸合成受抑制、细胞壁受损或膜面积受扰动时，PlsB的动态变化规律。此外，针对PlsB点突变菌株的宽场荧光成像实验，用于评估单个氨基酸替换如何影响蛋白聚集与膜结合特性。电子显微镜（EM）数据集则通过分析微细胞、超薄切片细胞以及不同制备条件下纯化的PlsB样品的超微结构特征，对上述观测结果进行补充。负染色与冷冻透射电子显微镜（cryo-TEM）成像可在高分辨率下评估PlsB潜在的寡聚化状态与膜相互作用情况。综上，荧光显微镜与电子显微镜数据集共同为解析PlsB的空间组织模式及其与膜生物发生的耦合关系提供了多尺度研究视角。此外，本数据集还包含成功制备囊泡的成像结果，未来可将其作为载体用于PlsB的体外研究。配套提供的MATLAB分析脚本可定量提取细胞几何特征与PlsB荧光焦点统计数据，保障该成像数据的解读可重复开展。