Four cell lines with different plasma membrane tensions (MCF10A, MCF7, MDA-MB-231, and PC-3 cells) adhered on the track-etched membrane. (Shiomi A. et al., Nat. commun. 2024)
Four cell lines (MCF10A, MCF7, MDA-MB-231, and PC-3 cells) were adhered on the track-etched membrane (Merck, VCTP04700) precoated with 100 µg/mL fibronectin (063-05591, FUJIFILM) at 37˚C for 1 day, fixed with 2.5% glutaraldehyde in 20 mM HEPES buffer (pH7.0) for 2 h, and processed by 5% HILEM IL1000 (Hitachi High-Tech) at room temperature for 30 min. Images were acquired using a field emission scanning electron microscope (SU8220; Hitachi High-Tech), working at an accelerating voltage of 2.0 kV with a secondary electron detector.
Akifumi Shiomi, Taikopaul Kaneko, Kaori Nishikawa, Arata Tsuchida, Takashi Isoshima, Mayuko Sato, Kiminori Toyooka, Kentaro Doi, Hidekazu Nishikii, Hirofumi Shintaku (2024) High-throughput mechanical phenotyping and transcriptomics of single cells., Nature communications, Volume 15, Number 1, pp. 3812
Published in 2024 May 17 (Electronic publication in May 17, 2024, midnight )
(Abstract) The molecular system regulating cellular mechanical properties remains unexplored at single-cell resolution mainly due to a limited ability to combine mechanophenotyping with unbiased transcriptional screening. Here, we describe an electroporation-based lipid-bilayer assay for cell surface tension and transcriptomics (ELASTomics), a method in which oligonucleotide-labelled macromolecules are imported into cells via nanopore electroporation to assess the mechanical state of the cell surface and are enumerated by sequencing. ELASTomics can be readily integrated with existing single-cell sequencing approaches and enables the joint study of cell surface mechanics and underlying transcriptional regulation at an unprecedented resolution. We validate ELASTomics via analysis of cancer cell lines from various malignancies and show that the method can accurately identify cell types and assess cell surface tension. ELASTomics enables exploration of the relationships between cell surface tension, surface proteins, and transcripts along cell lineages differentiating from the haematopoietic progenitor cells of mice. We study the surface mechanics of cellular senescence and demonstrate that RRAD regulates cell surface tension in senescent TIG-1 cells. ELASTomics provides a unique opportunity to profile the mechanical and molecular phenotypes of single cells and can dissect the interplay among these in a range of biological contexts.(MeSH Terms)