RESUMEN

We report the development of a widely-tunable femtosecond fiber laser system and its application for two-photon vision studies. The source is based on an Er-doped fiber laser with spectral shift up to 2150 nm, followed by a second harmonic generation module to generate a frequency-doubled beam tunable from 872 to 1075 nm. The source delivers sub-230 fs pulses with nearly-constant duration over the entire tuning range, with output powers between 0.68-1.24 mW, which corresponds to a pulse energy of 13.2-24.1 pJ. Such pulse energy is sufficient for employing a system for measurements of two-photon scotopic spectral sensitivity of two-photon vision in humans. The laser parameters allow for very efficient and safe two-photon stimulation of the human visual system, as proved by a good separation between one- and two-photon thresholds for wavelengths below 950 nm, which we have confirmed for 3 healthy subjects.

RESUMEN

BackgroundNoninvasive assessment of metabolic processes that sustain regeneration of human retinal visual pigments (visual cycle) is essential to improve ophthalmic diagnostics and to accelerate development of new treatments to counter retinal diseases. Fluorescent vitamin A derivatives, which are the chemical intermediates of these processes, are highly sensitive to UV light; thus, safe analyses of these processes in humans are currently beyond the reach of even the most modern ocular imaging modalities.MethodsWe present a compact, 2-photon-excited fluorescence scanning laser ophthalmoscope and spectrally resolved images of the human retina based on 2-photon excitation (TPE) with near-infrared light. A custom Er:fiber laser with integrated pulse selection, along with intelligent postprocessing of data, enables excitation with low laser power and precise measurement of weak signals.ResultsWe demonstrate spectrally resolved TPE fundus images of human subjects. Comparison of TPE data between human and mouse models of retinal diseases revealed similarity with mouse models that rapidly accumulate bisretinoid condensation products. Thus, visual cycle intermediates and toxic byproducts of this metabolic pathway can be measured and quantified by TPE imaging.ConclusionOur work establishes a TPE instrument and measurement method for noninvasive metabolic assessment of the human retina. This approach opens the possibility for monitoring eye diseases in the earliest stages before structural damage to the retina occurs.FundingNIH, Research to Prevent Blindness, Foundation for Polish Science, European Regional Development Fund, Polish National Agency for Academic Exchange, and Polish Ministry of Science and Higher Education.

Asunto(s)

Oftalmoscopios , Imagen Óptica , Retina , Enfermedades de la Retina , Animales , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Ratones , Ratones Noqueados , Persona de Mediana Edad , Retina/diagnóstico por imagen , Retina/metabolismo , Enfermedades de la Retina/diagnóstico por imagen , Enfermedades de la Retina/genética , Enfermedades de la Retina/metabolismo

RESUMEN

We report an investigation of dispersion management of an all-polarization-maintaining Er-fiber oscillator mode-locked via a nonlinear amplification loop mirror in a figure-nine cavity configuration with two output ports. The performance of the laser was investigated within the net cavity dispersion ranging from -0.034 ps2 to +0.006 ps2. We show that the spectral and temporal phase of the pulses at both figure-nine outputs have clearly different characteristics. One of the laser outputs provides pulses with significantly better quality; nonetheless, the rejection output also offers ultrashort pulses with broad spectra. Pulses as short as 79 fs with an energy of 83 pJ were generated directly from the laser in the near-zero dispersion regime.

RESUMEN

A femtosecond frequency-doubled erbium-doped fiber laser with an adjustable pulse repetition rate is developed and applied in two-photon excited fluorescence microscopy. The all-fiber laser system provides the fundamental pulse at 1560 nm wavelength with 22 fs duration for the second harmonic generation, resulting in 1.35 nJ, 60 fs pulses at 780 nm. The repetition rate is adjusted by a pulse picker unit built-in within the amplifier chain, directly providing transform-limited pulses for any chosen repetition rate between 1 and 12 MHz. We employed the laser source to drive a scanning two-photon excited fluorescence microscope for ex vivo rat skin and other samples' imaging at various pulse repetition rates. Due to compactness, ease of operation, and suitable pulse characteristics, the laser source can be considered as an attractive alternative for Ti:Sapphire laser in biomedical imaging.