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07.07.16 Gurdon Institute scientists reveal ultra-high resolution 3D imaging of "virtually any subcellular structure"

last modified Jul 13, 2016 03:00 PM
The culmination of a decade's research on high-resolution fluorescence imaging techniques, resulting in a dramatically increased imaging depth combined with nanoscale resolution, is presented in new Cell paper

Ultra-High Resolution 3D Imaging of Whole Cells

Huang F et al. (2016) Cell Jul 6. DOI: 10.1016/j.cell.2016.06.016. [Epub ahead of print]

Imaging experts George Sirinakis and Edward Allgeyer from the Gurdon Institute, working as part of a team of cell biologists, engineers and microbiologists drawn principally from departments of Yale University School of Medicine, describe their new technique of whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN). The scientists present exciting evidence to support their claim that this tool "opens the door to address cell biological questions that were previously unanswerable".

Fluorescence nanoscopy, or super-resolution microscopy, is an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50 to 80 nm) and rapidly deteriorating resolution in thick samples, its practical biological application has been effectively limited to two dimensions and thin samples.

Now, W-4PiSMSN allows imaging of three-dimensional (3D) structures at 10- to 20-nm resolution throughout entire mammalian cells. Refined hardware and new data analysis allow imaging of cells as thick as 10 μm, an improvement of up to 40-fold on previous similar imaging tools.

The ability to apply high resolution microscopy beyond small sub-volumes of cells and up to the extent of the whole cell will allow scientists to image whole organelles that span large volumes, and to probe the distribution of proteins across organelles such as endoplasmic reticulum, mitochondria, bacteriophages and nuclear pore complexes.

Sirinakis NuclearEnv  Sirinakis Cilium


Images reproduced from the paper's graphical abstract under Creative Commons license (CC BY 4.0).


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