Format

Send to

Choose Destination
Sci Rep. 2017 Oct 27;7(1):14185. doi: 10.1038/s41598-017-13793-3.

Dual Biomembrane Force Probe enables single-cell mechanical analysis of signal crosstalk between multiple molecular species.

Ju L1,2,3,4, Chen Y5,6,7, Li K8,5, Yuan Z5,6, Liu B8,5, Jackson SP9,10, Zhu C11,12,13.

Author information

1
Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, 30332, GA, United States. arnold.ju@sydney.edu.au.
2
Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, 30332, GA, USA. arnold.ju@sydney.edu.au.
3
Heart Research Institute, Newtown, 2050, NSW, Australia. arnold.ju@sydney.edu.au.
4
Charles Perkins Centre, The University of Sydney, Camperdown, 2006, NSW, Australia. arnold.ju@sydney.edu.au.
5
Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, 30332, GA, USA.
6
Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, 30332, GA, USA.
7
Department of Molecular Medicine, MERU-Roon Research Center on Vascular Biology, The Scripps Research Institute, La Jolla, 92037, CA, USA.
8
Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, 30332, GA, United States.
9
Heart Research Institute, Newtown, 2050, NSW, Australia.
10
Charles Perkins Centre, The University of Sydney, Camperdown, 2006, NSW, Australia.
11
Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, 30332, GA, United States. cheng.zhu@bme.gatech.edu.
12
Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, 30332, GA, USA. cheng.zhu@bme.gatech.edu.
13
Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, 30332, GA, USA. cheng.zhu@bme.gatech.edu.

Abstract

Conventional approaches for studying receptor-mediated cell signaling, such as the western blot and flow cytometry, are limited in three aspects: 1) The perturbing preparation procedures often alter the molecules from their native state on the cell; 2) Long processing time before the final readout makes it difficult to capture transient signaling events (<1 min); 3) The experimental environments are force-free, therefore unable to visualize mechanical signals in real time. In contrast to these methods in biochemistry and cell biology that are usually population-averaged and non-real-time, here we introduce a novel single-cell based nanotool termed dual biomembrane force probe (dBFP). The dBFP provides precise controls and quantitative readouts in both mechanical and chemical terms, which is particularly suited for juxtacrine signaling and mechanosensing studies. Specifically, the dBFP allows us to analyze dual receptor crosstalk by quantifying the spatiotemporal requirements and functional consequences of the up- and down-stream signaling events. In this work, the utility and power of the dBFP has been demonstrated in four important dual receptor systems that play key roles in immunological synapse formation, shear-dependent thrombus formation, and agonist-driven blood clotting.

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center