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Manchester Academic Health Science Centre

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Enrichment of Bruch's Membrane from Human Donor Eyes

Selina McHarg^1,2, Nicole Brace^1,2, Paul N. Bishop^1,2, Simon J. Clark^1,2

¹Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester, ²Centre for Advanced Discovery and Experimental Therapeutics, Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre

Identifying proteins specifically associated with Bruch’s membrane in human eyes is an important step in understanding the biochemical mechanisms behind eye diseases such as age-related macular degeneration. This protocol describes how to enrich this sheet of extracellular matrix for down-stream biochemical analysis.

Neuroscience

Light-Induced Molecular Adsorption of Proteins Using the PRIMO System for Micro-Patterning to Study Cell Responses to Extracellular Matrix Proteins

Cristina Melero^*1, Aljona Kolmogorova^*1, Paul Atherton¹, Brian Derby², Adam Reid^3,4, Karin Jansen⁵, Christoph Ballestrem¹

¹Faculty of Biology, Medicine and Health. Division of Cell Matrix, Biology and Regenerative Medicine, Wellcome Trust Centre for Cell-Matrix Research, The University of Manchester, ²School of Materials, The University of Manchester, ³Blond McIndoe Laboratories, School of Biological Sciences, Faculty of Biology, Medicine and Health, Division of Cell Matrix Biology and Regenerative Medicine, The University of Manchester, Manchester Academic Health Science Centre, ⁴Department of Plastic Surgery & Burns, Wythenshawe Hospital, Manchester University NHS Foundation Trust, Manchester Academic Health Science Centre, ⁵Department of Pathology, UMC Utrecht

Our overall aim is to understand how cells sense extracellular cues that lead to directed axonal growth. Here, we describe the methodology of Light-Induced Molecular Adsorption of Proteins, used to produce defined micro-patterns of extracellular matrix components in order to study specific events that govern axon outgrowth and pathfinding.