Heart Research Institute

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

In Vitro Stimulation and Visualization of Extracellular Trap Release in Differentiated Human Monocyte-derived Macrophages

Yunjia Zhang^1,2, Benjamin S. Rayner^1,2, Mathias Jensen³, Clare L. Hawkins^1,2,3

¹Heart Research Institute, ²Sydney Medical School, University of Sydney, ³Department of Biomedical Sciences, University of Copenhagen

Presented here is a protocol to detect macrophage extracellular trap (MET) production in live cell culture using microscopy and fluorescence staining. This protocol can be further extended to examine specific MET protein markers by immunofluorescence staining.

Bioengineering

Molecular Spring Constant Analysis by Biomembrane Force Probe Spectroscopy

Peyman Obeidy¹, Haoqing Wang^1,2,3, Mingqin Du¹, Huiqian Hu^1,4, Fang Zhou¹, Haoruo Zhou⁵, Hao Huang⁵, Yunduo Charles Zhao^1,2, Lining Arnold Ju^1,2,3

¹School of Biomedical Engineering, Faculty of Engineering, The University of Sydney, ²Charles Perkins Centre, The University of Sydney, ³Heart Research Institute, ⁴Department of Chemistry, The Hong Kong University of Science and Technology, ⁵School of Aerospace, Mechanical and Mechatronic Engineering, Faculty of Engineering, The University of Sydney

A biomembrane force probe (BFP) is an in situ dynamic force spectroscopy (DFS) technique. BFP can be used to measure the spring constant of molecular interactions on living cells. This protocol presents spring constant analysis for molecular bonds detected by BFP.

Biology

Fluorescence Micropipette Aspiration Assay to Investigate Red Blood Cell Mechanosensing

Jasmine Jin¹, Haoqing Jerry Wang^1,2,3, Yiyao Catherine Chen¹, Blake Russell¹, Allan Sun^1,2,3,4, Yao Wang¹, Lining Arnold Ju^1,2,3,4

¹School of Biomedical Engineering, The University of Sydney, ²Charles Perkins Centre, The University of Sydney, ³Heart Research Institute, ⁴The University of Sydney Nano Institute (Sydney Nano), The University of Sydney

The exploration of cellular behavior under mechanical stress is pivotal for advances in cellular mechanics and mechanobiology. We introduce the Fluorescence Micropipette Aspiration (fMPA) technique, a novel method combining controlled mechanical stimulation with comprehensive analysis of intracellular signaling in single cells. This technique investigates new in-depth studies of live-cell mechanobiology.