Increased signals from short-wavelength-excited fluorescent molecules using sub-Ti:Sapphire wavelengths

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Norris, G and Amor, R and Dempster, J and Amos, W B and McConnell, G (2012) Increased signals from short-wavelength-excited fluorescent molecules using sub-Ti:Sapphire wavelengths. Journal of Microscopy, 248 (2). pp. 200-207. ISSN 0022-2720 (https://doi.org/10.1111/j.1365-2818.2012.03663.x)

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Abstract

We report the use of an all-solid-state ultrashort pulsed source specifically for two-photon microscopy at wavelengths shorter than those of the conventional Ti:Sapphire laser. Our approach involves sum-frequency mixing of the output from an optical parametric oscillator (λ= 1400-1640 nm) synchronously pumped by a Yb-doped fibre laser (λ= 1064 nm), with the residual pump radiation. This generated an fs-pulsed output tunable in the red spectral region (λ= 620-636 nm, ~150 mW, 405 fs, 80 MHz, M(2) ~ 1.3). We demonstrate the performance of our ultrashort pulsed system using fluorescently labelled and autofluorescent tissue, and compare with conventional Ti:Sapphire excitation. We observe a more than 3-fold increase in fluorescence signal intensity using our visible laser source in comparison with the Ti:Sapphire laser for two-photon excitation at equal illumination peak powers of 1.16 kW or less.

ORCID iDs

Norris, G, Amor, R, Dempster, J ORCID logoORCID: https://orcid.org/0000-0003-2199-2945, Amos, W B and McConnell, G ORCID logoORCID: https://orcid.org/0000-0002-7213-0686;
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