Centre for Optical Instrumentation Laboratory (COIL)

Biotechnology
Genotype to phenotype
Centre for Optical Instrumentation Laboratory (COIL)
Project contact
Cameron Chalmers
Consultancy Associate
Cameron.Chalmers@ei.ed.ac.uk

High-resolution analysis of cell division errors with COIL

The JP group at the University of Edinburgh studies cell division, the essential biological process which ensures genome integrity by equally and identically distributing chromosomes between two daughter cells. Errors in cell division often result in daughter cells with inappropriate chromosome numbers, a condition associated with numerous cancers and birth defects. Although much is known about the basic mechanisms of cell division, structural level mechanistic details of the pathways regulating error-free chromosome segregation are still emerging. In particular, a high-resolution understanding of centromere inheritance and how kinetochores employ dynamic protein interactions to drive chromosome segregation is yet to be obtained.

Dr Maria Alba Abad Fernandez, a postdoc within the JP group, has been studying part of this process – specifically the interaction between the chromosomal passenger complex (CPC) and Shugoshin 1 (Sgo-1), two major regulators of cell division. CPC and Sgo-1 directly interact and need to associate with centromeres to perform their function but the minimal region of Sgo-1 required for their direct binding and how this influences CPC centromere localisation and function were still unknown.

Working with Dr David Kelly within the COIL facility, Dr Abad used cell-based in vivo functional assays and high-resolution microscopy with automated analysis programmes to assess the effect of a number of Sgo-1 mutants on Sgo-1’s ability to bind CPC. Critically, David developed a bespoke ImageJ plugin for the study which enabled Dr Abad to automate the analysis of micrographs and identify the key regions of Sgo-1 required for this interaction in cells. Through this study, Dr Abad discovered that the N-terminus of Sgo-1 is essential for Sgo-1-CPC assembly and that, without this Sgo-1 region, the interaction is disrupted causing perturbed CPC centromere localisation and function, which leads to chromosome missegregation.

Centre for Optical Instrumentation Laboratory (COIL)

Centre for Optical Instrumentation Laboratory

Specialisms

  • Agri-tech, agri-food and plant biology
  • Food and drink
  • Healthcare and disease
  • Life sciences and industrial biotechnology
  • Pharmaceuticals and medical biotechnology


Technologies available

The COIL facility houses a range of microscopes capable of performing timelapse imaging at high temporal and spatial resolutions whilst maintaining focus. These can examine specimens from yeast and fungus through to embryos and polymer lattices. The facility staff offer their expertise and guidance to ensure researchers use the correct setup to achieve their desired outcome and are adept at writing bespoke ImageJ plugins to create analysis pipelines. These can assist by speeding up the analysis process or adding functionality to the existing software.


Equipment available

Microscopes capable of performing timelapse imaging at high temporal and spatial resolutions whilst maintaining focus.

  • Zeiss LSM 980 Airyscan Confocal
  • Zeiss LSM 880 Airyscan confocal
  • Leica SP5 laser scanning confocal microscope
  • Zeiss TIRF III
  • Nikon Ti2 CSU-W1 Spinning Disk Confocal
  • Nikon Ti2 widefield microscope
  • Zeiss Axio Observer widefield microscope
  • Deltavision Elite widefield microscope
  • Thermo Fisher Attune NxT Flow Cytometer


Software available

Image Analysis/Processing

  • Imaris#
  • ImageJ/Fiji
  • Metamorph
  • Zeiss Zen 3.0


More details

Centre for Optical Instrumentation Laboratory

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