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The Babraham Institute

3 ARTICLES PUBLISHED IN JoVE

Biology

Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond

Eleftherios Karanasios¹, Eloise Stapleton², Simon A. Walker¹, Maria Manifava¹, Nicholas T. Ktistakis¹

¹Signalling Programme, The Babraham Institute, ²MRC Group, Cardiff School of Biosciences, Cardiff University

Time-lapse microscopy of fluorescently labeled autophagy markers allows monitoring of the dynamic autophagy response with high temporal resolution. Using specific autophagy and organelle markers in a combination of 3 different colors, we can follow the contribution of a protein to autophagosome formation in a robust spatial and temporal context.

Biology

Robust 3D DNA FISH Using Directly Labeled Probes

Daniel J. Bolland¹, Michelle R. King², Wolf Reik^2,3, Anne E. Corcoran¹, Christel Krueger²

¹Nuclear Dynamics Programme, The Babraham Institute, ²Epigenetics Programme, The Babraham Institute, ³Centre for Trophoblast Research, University of Cambridge

We describe a robust and versatile protocol for analyzing nuclear architecture by 3D DNA FISH using directly labeled fluorescent probes.

Biology

Evaluating Autophagy Levels in Two Different Pancreatic Cell Models Using LC3 Immunofluorescence

Felipe J. Renna¹, Malena Herrera Lopez¹, Maria Manifava², Nicholas T. Ktistakis², Maria I. Vaccaro¹

¹Instituto de Bioquimica y Medicina Molecular Prof Alberto Boveris (IBIMOL), Universidad de Buenos Aires, CONICET, ²Signalling Programme, The Babraham Institute

The goal of this protocol is to determine autophagic levels in pancreatic cancer and pancreatic acinar cells through LC3 immunofluorescence and LC3 dot quantification.