Single Molecule

Single-molecule fluorescence (SMF) imaging has emerged as a complementary structural biology technique to enable high-resolution measurements of protein dynamics, mechanism and function. In particular, SMF microscopy is a highly specific, and non-invasive imaging method, capable of visualising individual fluorophores conjugated to biological molecules of interest.

Advantages of the SMF approaches include:
1) Protein dynamics; measurements of (millisecond) dynamics, in real-time, and with nanometre resolution in the range of 1-10 nm
2) High-contrast; only those proteins bearing a label are visualised
3) Single-molecule statistics; probe rare events and states that would otherwise be obscured in standard bulk measurements.
4) High-resolution; visualise structures below the diffraction limit (~250 nm) of conventional optical microscopy.

The laboratory of Dr Alan Lowe has constructed apparatus with the following capabilities:

1) Single-molecule Forster Resonance Energy Transfer (smFRET)
2) Single-molecule Tracking
3) Super-resolution Localisation Microscopy (dSTORM and PALM)
4) Timelapse imaging
5) Total Internal Reflection Fluorescence (TIRF) imaging

Researchers interested in using these instruments should contact Dr Alan Lowe.

In addition, the laboratory of Dr Anthony Roberts has a Nikon H-TIRF system for single-molecule analysis, equipped with a four colour laser launch and EMCCD camera.

These instruments for single-molecule measurements are complemented by ISMB microscopes for cellular imaging, including a Leica SP5 confocal microscope and Zeiss microscope for epifluorescence.