荧光显微镜是细胞生物学和癌症研究的重要工具。目前实施先进光学显微镜方法与细胞中特定蛋白质的免疫荧光标记相结合的方法现在能够提供高达 ∼25 nm 的光学超分辨率。在这里，我们使用标准免疫染色方案结合简易随机光学重建显微镜（easySTORM）进行超分辨成像，以观察三种不同细胞类型（人骨髓间充质干细胞（MSC））中两种细胞骨架元件肌动蛋白和微管蛋白的结构差异、人胶质母细胞瘤 (U87MG) 和乳腺癌 (MDAMB-231) 细胞。从干细胞的 STORM 图像中获得的肌动蛋白束的平均宽度被发现大于 U87MG 和 MDAMB-231 细胞的平均宽度。然而，相同细胞内微管蛋白的宽度没有显着差异。我们还研究了 NICD1 和 β-catenin 沉默的 MDAMB-231 细胞对 2D 迁移潜力的功能影响。尽管与对照细胞相比，具有上述两种条件的细胞观察到相似的迁移速度，但 easySTORM 图像显示，β-连环蛋白沉默的 MDAMB-231 细胞中肌动蛋白的宽度明显低于对照细胞中的肌动蛋白宽度。然而，这种微小的差异在宽视场图像中是观察不到的。我们的 easySTORM 研究结果应该有利于研究人员对细胞结构和潜在的治疗应用进行详细研究。© 2024 作者。由 Elsevier B.V. 出版

Fluorescence microscopy is an important tool for cell biology and cancer research. Present-day approach of implementing advanced optical microscopy methods combined with immunofluorescence labelling of specific proteins in cells is now able to deliver optical super-resolution up to ∼25 nm. Here we perform super-resolved imaging using standard immunostaining protocol combined with easy stochastic optical reconstruction microscopy (easySTORM) to observe structural differences of two cytoskeleton elements, actin and tubulin in three different cell types namely human bone marrow-derived mesenchymal stem cells (MSCs), human glioblastoma (U87MG) and breast cancer (MDAMB-231) cells. The average width of the actin bundle obtained from STORM images of stem cells is observed to be larger than the same for U87MG and MDAMB-231 cells. No significant difference is however noticed in the width of the tubulin within the same cells. We also study the functional effect on the 2D migration potential of MDAMB-231 cells silenced for NICD1 and β-catenin. Although similar migration speed is observed for cells with the above two conditions compared to their control cells, easySTORM images show that widths of the actin in MDAMB-231 cells in β-catenin silenced is significantly lower than the same in control cells. Such minute differences however are not observable in widefield images. The outcome of our easySTORM investigation should benefit the researchers carrying out detailed investigations of the cellular structure and potential therapeutic applications.© 2024 The Authors. Published by Elsevier B.V.