3D Photoactivated Localisation Microscopy (PALM) & Stochastic Optical Reconstruction Microscopy (STORM)

funded by a kind gift from Michael Beverley – Alumnus (Economics & Politics 1973) and campaign board member for the University’s Making a World of Difference campaign

Capabilities

Fluorescence localisation microscopy (3D PALM/STORM) for 20 nm precision of label (fluorescent protein tag or conjugated antibody) in fixed samples.

Choice of five excitation wavelengths.

Lasers

  • 405 nm
  • 488 nm
  • 532 nm
  • 561 nm
  • 647 nm

Objective

  • Water 60x objective, NA 1.20 (UPlanSApo)

Location

Miall 5.07, Faculty of Biological Sciences

Contacts

Dr Alistair Curd
a.curd@leeds.ac.uk
0113 34 37208

For further information about using the microscope, training and applications - please contact Alistair

Please note this microscope is currently free to use whilst it is still in development.

Some examples of images taken using STORM and PALM so far...

Adult rat cardiomyocyte expressing an Eos-labelled myopalladin construct. Myopalladin tethers various proteins to the z-discs of sarcomeres in muscle cells. Top; example of 200 frames of data capture, which (after further rounds of data capture) can be reconstructed to give an image such as Bottom; Eos-myopalladin localising at z-discs.

Adult rat cardiomyocyte expressing an Eos-labelled Actinin construct. Actinin is a z-disc interacting protein that organises actin filaments. Scale bar 2 µm.

Publications

Bartlett, C. et al. (2015) Developing tools to study hepatitis C virus replication using super-resolution microscopy, Society for General Microbiology Annual Conference 2015, Abstract S12We0948

Lambacher, NJ et al (2016) TMEM107 recruits ciliopathy proteins to anchored periodic subdomains of the ciliary transition zone membrane and is mutated in Joubert syndrome. Nature Cell Biology, 18, 122–131

Links & protocols

PALM instructions

Nikon STORM Protocols