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

• Published in: Biochemistry and biophysics reports 2024-09, Vol.39, p.101798, Article 101798
• Main Authors: Bharadwaj, Anupam, Kumar, Amalesh, Padalumavunkal Mathew, Sam, Mitra, Rumela, Bhattacharyya, Jina, Jaganathan, Bithiah Grace, Boruah, Bosanta R.
• Format: Article
• Language: English
• Subjects: actin cytoskeleton; Breast cancer; Optical super-resolution microscopy; Short Communication; Stochastic optical reconstruction microscopy; Tubulin; β-catenin
• ISSN: 2405-5808; 2405-5808
• DOI: 10.1016/j.bbrep.2024.101798

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. •Super-resolution based on easySTORM of MSC, U87MG and MDAMB-231 cell lines.•Actin bundle width in MSC appears larger than in U87MG and MDAMB-231 cells.•Estimated minimum width of actin bundles in the breast cancer cell line is ∼32 nm.•Tubulin in glioblastoma and breast cancer cell lines shows minimum width as 24 nm.•easySTORM shows a ∼25 % decrease in actin bundle width in β-catenin silenced cells.