Technical information and user guides

Quick start and single channel dSTORM guide

dSTORM analysis

ONI website

Location: MRC Building. Room G.10.

ONI Nanoimager S single molecule localisation microscopy (SMLM) system designed for smFRET and single molecule tracking. Fill in this brief web form if you are interested in using the machine or would like more information

Recent publications using the Nanoimager

In the preprint below, Georgiou and Cabella-Garcia et al used two-colour direct stochastic optical reconstruction microscopy (dSTORM) to examine the binding selectivity of engineered DNA-nanostructures to unilamellar vesicles of different sizes and lipid membrane curvatures

Georgiou E., Cabella-Garcia J., Xing Y. & Howorka S. DNA Origami – Lipid Membrane Interactions Defined at Single-Molecular Resolution. bioRχiv (2023). https://doi.org/10.1101/2023.11.14.567022

Alanazi et al describe how to use single molecule Förster Resonance Energy Transfer (smFRET) to study the conformation states of DNA Holliday Junctions on a molecule-by-molecule basis. A technique that can be extrapolated for studying the dynamic behaviour of other nanostructures without the averaging effect of ensemble measurements.

Alanazi, A.F.R., Haider, S. & Parkinson, G.N. The use of single-molecule FRET for the characterization of Holliday junctions containing human telomeric DNA: a methodological approach for nanoscale distance, mobility, and stability measurements. Med Chem Res (2024). https://doi.org/10.1007/s00044-024-03197-0

About the Nanoimager

The CMOS camera can be used to simultaneously acquire images from two fluorescence channels. The beam-splitter sends far red light to one channel and blue, green or orange light to the other. Ideal fluorophores are Cy3 and Cy5, or dyes with similar spectra. Note that the system is optimised for the Cy3/Cy5 FRET pair and it is not possible to simultaneously image samples labelled with both green and red fluorophores (e.g. EGFP and mCherry).

Other advantages of the system are the speed and field of view of the CMOS camera and built-in microfluidics. The analysis software is freely available for installation (Windows 10 PC required).

Use for:

Simultaneous dual-channel single molecule localisation microscopy
smFRET
Single molecule tracking
Microfluidics for single molecule imaging

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