NPL is active in the development and application of super-resolution microscopy for a broad range of measurement applications. We have developed two bespoke platforms for super-resolution imaging at the highest spatial and temporal resolution. The open architecture of both the hardware and software allows us to optimise the systems for specific applications.
- direct Stochastic Optical Reconstruction Microscopy (dSTORM) offers the highest spatial resolution achievable using far field light microscopy, with practical localisation precision down to 10-15 nm. The technique relies on detecting the positions of single fluorescent molecules and particular care is required in the labelling and preparation of samples to achieve good results. Image acquisition time is relatively slow, which typically limits the effective use of dSTORM to fixed samples.
- Structured Illumination Microscopy (SIM) is a fast, widefield super-resolution imaging technique in which the sample is illuminated using spatially patterned light. The NPL SIM is capable of multi-colour 3D imaging with a lateral resolution down to ~ 100 nm and is compatible with most common fluorescent dyes and proteins. The high imaging speed makes it well suited to imaging dynamic system such as live cells.
We have extensive experience in the preparation and labelling of wide range of samples for super-resolution imaging, including live and fixed cells prepared in our in-house cell culture facilities. We also operate a laser scanning confocal microscope for diffraction-limited imaging.
A two-tier Golgi-based control of organelle size underpins the functional plasticity of endothelial cells
Ferraro F, Kriston-Vizi J, Metcalf D J, Martin-Martin B, Freeman J, Burden J J, Westmoreland D, Dyer C E, Knight A E, Ketteler R and Cutler D F
Dev Cell, 29, 292-304 (2014)
TestSTORM: simulator for optimizing sample labeling and image acquisition in localization based super-resolution microscopy
Sinkó J, Kákonyi R, Rees E, Metcalf D, Knight A E, Kaminski C F, Szabó G and Erdélyi M
Biomed Opt Express, 5, 778-787 (2014)
Elements of image processing in localization microscopy
Eric J Rees, Miklos Erdelyi, Gabriele S Kaminski Schierle, Alex Knight and Clemens F Kaminski
Journal of Optics, 15 (9):094012 (2013)
Test Samples for Optimizing STORM Super-Resolution Microscopy
Metcalf, D., Edwards, R., Kumarswami, N., Knight, A.
J. Vis. Exp., 79, e50579, doi:10.3791/50579 (2013)
Correcting chromatic offset in multicolor super-resolution localization microscopy
Erdelyi M, Rees E, Metcalf D, Schierle GS, Dudas L, Sinko J, Knight AE, Kaminski CF
Opt Express, 21, 10978-10988 (2013)
Blind assessment of localisation microscope image resolution
Rees E, Erdelyi M, Pinotsi D, Knight AE, Metcalf D, Kaminski CF
Optical Nanoscopy,1.12, 2012
Single Molecule Pointillism
Erdelyi M, Metcalf D
Imaging & Microscopy (2012)
In Situ Measurements of the Formation and Morphology of Intracellular β-Amyloid Fibrils by Super-Resolution Fluorescence Imaging
Schierle GSK, van de Linde S, Erdelyi M, Esbjorner EK, Klein T, Rees T, Bertoncini CW, Dobson CM, Sauer M, Kaminski CF
J. Am. Chem. Soc., 133 (33), pp 12902-12905 (2011)
Super-resolution microscopy as a potential approach to diagnosis of platelet granule disorders
Westmoreland, D., Shaw, M., Grimes, W., Metcalf, D. J., Burden, J. J., Gomez, K., Knight, A.E. and Cutler, D.F.
J. Thromb. Haemost., 13:1-11 (2016)
High speed structured illumination microscopy in optically thick samples
Shaw, M., et al
Methods, 88 (2015)
Filming protein fibrillogenesis in real time
Bella, A., et al.
Sci. Rep., 4 (2014)
Optimized approaches for optical sectioning and resolution enhancement in structured illumination microscopy
O'Holleran, K., Shaw, M
Biomed. Opt. Express, 5 (2014)
Polarization effects on contrast in structured illumination microscopy
O'Holleran K, Shaw MJ
Opt. Lett., 37, 4603-4605 (2012)
- Correlative super resolution using dSTORM and SIM
- STORM-ing through the diffraction limit
- Application of Super-Resolution Imaging to the Endocytic Pathway
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