Recent advances in RESOLFT (reversible saturable optical fluorescence transitions) microscopy have enabled the non-invasive three-dimensional visualization of numerous structures in living cells at high spatial resolution. This technique, which utilizes low light intensities, has manifold potential applications in the life sciences. The work presented here envisions further broadening the applications of RESOLFT microscopy by implementing a scheme for fast image acquisition of large fields of view, applicable to thick specimen.
With a pattern consisting of line-shaped intensity minima, this novel technique, called lineRESOLFT, permits fast imaging of living cells at ~40nm lateral resolution while offering strong optical sectioning. The full potential of this method is further illustrated by the achievement of continuous three-dimensional imaging of neurons in living brain slices with high spatio-temporal resolution, enabling the observation of rapid spine motility for large fields of view on the second time scale
