Advancing cellular insights: Super-resolution STORM imaging of cytoskeletal structures in human stem and cancer cells

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2024-07-25 DOI:10.1016/j.bbrep.2024.101798

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Abstract

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.

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增进对细胞的了解：人类干细胞和癌细胞细胞骨架结构的超分辨率 STORM 成像

荧光显微镜是细胞生物学和癌症研究的重要工具。如今，通过采用先进的光学显微镜方法，结合对细胞中特定蛋白质的免疫荧光标记，可实现高达 25 纳米的光学超分辨。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，观察三种不同类型细胞（即人骨髓间充质干细胞（MSCs）、人胶质母细胞瘤（U87MG）和乳腺癌（MDAMB-231）细胞）中两种细胞骨架元素（肌动蛋白和微管蛋白）的结构差异。从 STORM 图像中获得的干细胞肌动蛋白束平均宽度大于 U87MG 和 MDAMB-231 细胞。不过，在相同细胞内，微管蛋白的宽度没有明显差异。我们还研究了沉默NICD1和β-catenin的MDAMB-231细胞对二维迁移潜力的功能影响。虽然在上述两种条件下观察到的细胞迁移速度与对照细胞相似，但easySTORM图像显示，沉默β-catenin的MDAMB-231细胞的肌动蛋白宽度明显低于对照细胞。但这种微小的差异在宽场图像中无法观察到。我们的 EasySTORM 研究结果将有助于研究人员对细胞结构和潜在治疗应用进行详细研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.