Out-of-phase imaging after optical monochromatic modulation (OPIOMM) enables the selective and quantitative imaging of a photoswitchable fluorophore in presence of interfering species emitting in the same wavelength range. This simple approach relies on a modulated monochromatic light excitation and a kinetic filter based on matching the average intensity and frequency of the modulated excitation to the dynamics of the photoswitchable probe. OPIOMM enabled selective imaging in mammalian cells and zebrafish (see Figure).Upon using photoswitchable fluorescent proteins1 or photoswitchable fluorogenic hybrids,2this imaging protocol will be especially useful to overcome autofluorescence and light scattering. It will also allow us to achieve highly multiplexed observations upon adding a temporal dimension to spectral discrimination.References: 1. J. Querard, T. Markus, M.-A. Plamont, C. Gauron, P. Wang, A. Espagne, M. Volovitch, S. Vriz, V. Croquette, A. Gautier, T. Le Saux, L. Jullien, submitted; 2. P. Wang, J. Querard, S. Maurin, S. S. Nath, T. Le Saux, A. Gautier, L. Jullien, Chem. Sci., 2013, 4, 2865.

Out-of-phase imaging after optical monochromatic modulation

248th National Meeting (2014)

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