Abstract
Multi-photon microscopy is a powerful tool in biomolecular research. Less complex and more cost effective excitation light sources will make this technique accessible to a broader community. Semiconductor diode seeded fiber lasers have proven to be especially robust, low cost and easy to use. However, their wavelength tuning range is often limited, so only a limited number of fluorophores can be accessed. Therefore, different approaches have been proposed to extend the spectral coverage of these lasers. Recently, we showed that four-wave mixing (FWM) assisted stimulated Raman scattering (SRS) can be harnessed to red-shift high power pulses from 1064 nm to a narrowband output at 1122 nm and 1186 nm and therefore extend the number of accessible fluorophores. In this contribution, we show the applicability of all three wavelengths for multi-photon microscopy and analyze the performance.
Highlights
Non-linear biomolecular imaging is a growing field with a strong demand for high power lasers
We presented a simple and straightforward way to use four-wave mixing (FWM) seeded stimulated Raman scattering (SRS) for extending the spectral coverage of single wavelength fiber lasers
We presented two-photon excitation fluorescence (TPEF) imaging of plants and second harmonic generation (SHG) imaging of mouse skin at 1064 nm excitation
Summary
Non-linear biomolecular imaging is a growing field with a strong demand for high power lasers. Two-photon excitation fluorescence (TPEF) microscopy is an easy-to-use and widely deployed imaging technique [1]. Compared to other optical 3D imaging modalities, like for example optical coherence tomography (OCT) [2], it has the advantage of good molecular specificity. Either due to auto-fluorescence or by marking specific sites with a fluorophore, it is ensured that only the molecular structures of interest are visible in the image and the surrounding areas remain dark. Together with its sub-cellular spatial resolution this makes TPEF microscopy a valuable tool for researchers in fields like biomolecular or neuronal science [3,4,5]
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