A laser-based “green” synthesis of nanoparticles (NPs) was used to manufacture gold NPs in water. The light source is a Ti:Sapphire laser with 30 fs FWHM pulses, 800 nm mean wavelength, and 1 kHz repetition rate. The method involves two stages: (1) pulsed laser ablation in liquids and (2) photo-fragmentation (PF). Highly pure and well-dispersed NPs with a diameter of 18.5 nm that can be stored at room temperature without showing any agglomeration over a period of at least 3 months were produced without the need to use any stabilizer. Transmittance spectra, extinction coefficient, NPs agglomeration dynamics, and thermal conductivity of the nanofluids obtained were analyzed before and after being submitted to thermal cycling and compared to those obtained for commercial gold/water suspensions. Optical properties have also been investigated, showing no substantial differences for thermal applications between NPs produced by the laser ablation and PF technique and commercial NPs. Therefore, nanofluids produced by this technique can be used in thermal applications, which are foreseen for conventional nanofluids, e.g., heat transfer enhancement and solar radiation direct absorption, but offering the opportunity to produce them in situ in almost any kind of fluid without the production of any chemical waste.