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Queen's University Belfast

4 ARTICLES PUBLISHED IN JoVE

Biology

Preparation of Highly Coupled Rat Heart Mitochondria

¹Faculty of Life Sciences, University of Manchester, ²School of Biological Sciences, Queen's University Belfast

We describe а protocol for isolation of pure, highly coupled rat heart mitochondria for functional or structural studies of cellular bioenergetics, biophysical measurements, proteomics or mitochondrial DNA and lipids analysis.

Biochemistry

Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions

Myriam Jaraíz-Rodríguez¹, Ana González-Sánchez^1,2, Laura García-Vicente¹, Jose M. Medina¹, Arantxa Tabernero¹

¹Instituto de Neurociencias de Castilla y León (INCYL), Departamento de Bioquímica y Biología Molecular, Universidad de Salamanca, ²Centre for Cancer Research & Cell Biology (CCRCB), School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast

This is a protocol to study intracellular protein-protein interactions based on the biotin-avidin pull-down system with the novelty of combining cell-penetrating sequences. The main advantage is that the target sequence is incubated with living cells instead of cell lysates and therefore the interactions will occur within the cellular context.

Bioengineering

Microfluidic Production of Lysolipid-Containing Temperature-Sensitive Liposomes

Calvin C. L. Cheung¹, Guanglong Ma¹, Amalia Ruiz¹, Wafa T. Al-Jamal¹

¹School of Pharmacy, Queen's University Belfast

The protocol presents the optimized parameters for preparing thermosensitive liposomes using the staggered herringbone micromixer microfluidics device. This also allows co-encapsulation of doxorubicin and indocyanine green into the liposomes and the photothermal-triggered release of doxorubicin for controlled/triggered drug release.

Medicine

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Michael A. Pearen^*1, Hong Kiat Lim^*1, Francis D. Gratte^2,3, Manuel A. Fernandez-Rojo^1,4,5, Sujeevi K. Nawaratna⁶, Geoffrey N. Gobert⁷, John K. Olynyk^8,9, Janina E. E. Tirnitz-Parker^2,10, Grant A. Ramm^1,4

¹Hepatic Fibrosis Group, QIMR Berghofer Medical Research Institute, ²School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, ³School of Veterinary and Life Sciences, Murdoch University, ⁴School of Medicine, The University of Queensland, ⁵Madrid Institute for Advanced Studies (IMDEA) in Food, CEI UAM+CSIC, ⁶School of Medicine, Griffith University, ⁷School of Biological Sciences, Queen's University Belfast, ⁸Department of Gastroenterology & Hepatology, Fiona Stanley and Fremantle Hospital Group, ⁹School of Medical and Health Sciences, Edith Cowan University, ¹⁰Centre for Cell Therapy and Regenerative Medicine, School of Biomedical Sciences, University of Western Australia

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.