Two-photon vision is a newly discovered mechanism of the perception of pulsed near-infrared laser beams as a color stimulus corresponding to approximately half of the laser wavelength (PNAS 111(50), E5445–E5454). Based on this phenomenon, a new visual field test instrumentation – two-photon microperimetry, has been developed (BOE 10(9), 4551–4567). This study shows that two-photon perimetry gives more reproducible results than one-photon perimetry for standard threshold finding strategies. This unquestionable advantage of the nonlinear vision-based visual field testing technique may benefit the clinical assessment of retinal disease progression and treatment efficiency.
Two-photon vision relies on the perception of pulsed near-infrared laser beams as having colors like their half-wavelength counterparts. The phenomenon is due to two-photon absorption occurring in visual pigments [1]. This study is focused on methods to determine the contrast sensitivity function (CSF) for two-photon vision, which has not yet been investigated. CSF was measured for eight spatial frequencies using the tumbling E letter optotype. The optotype was projected on a white background by fast scanning the retina with a pulsed 1040 nm or 520 nm laser beam, both perceived as green. The contrast threshold was determined for the power of the beam corresponding to a minimum stimulus brightness for which the subject was able to state the correct letter orientation. Because a luminance curve for the two-photon stimulus is not available, expressing the brightness of the infrared stimulus in photometric units required finding a suitable method. Three approaches for determining contrast sensitivity for two-photon stimulus were proposed and tested to overcome this problem. The threshold contrast values, defined as Weber contrast, differ substantially between normal and two-photon vision mechanisms. Each tested method allowed qualitative comparison of the obtained contrast sensitivities. The results show that the two-photon CSF has a significantly broader range than the one-photon CSF. Determining the CSF for twophoton vision will help assess the applicability of this phenomenon to augmented reality displays.
In this paper, we present the preliminary results of the scotopic luminosity curve for two-photon vision measurements in the spectral range from 872 nm to 1027 nm. The results were obtained thanks to a newly-developed custom-build tunable femtosecond erbium-doped fiber laser that pulse train parameters and spectral width are close to constant while tuning. Such instrumentation enabled us to perform reliable measurements across the laser tuning range of over 150 nm.
As reported previously, OCT sources emit short infrared pulses that may be seen due to two-photon vision. In this work, we quantified visibility of OCT sources, which may be beneficial for eye imaging system designers.
Pulsed near-infrared (NIR) light sources can be successfully applied for both imaging and functional testing of the human eye, as published recently. These two groups of applications have different requirements. For imaging applications, the most preferable is invisible scanning beam while efficiently visible stimulating beam is preferable for functional testing applications. The functional testing of human eye using NIR laser beams is possible due to two-photon vision (2PV) phenomenon. 2PV enables perception of pulsed near-infrared laser light as color corresponding to approximately half of the laser wavelength. This study aims to characterize two-photon vision thresholds for various pulse lengths from a solidstate sub-picosecond laser (λc = 1043.3 nm, Frep = 62.65 MHz), either of 253 fs duration or elongated by Martinez- type stretcher to 2 ps, and fiber-optic picosecond laser (λc = 1028.4 nm, Frep = 19.19 MHz, τp = 12.2 ps).
Development of new microperimetric tools dedicated for imaging of early functional changes in the retina may help in the monitoring of various ocular diseases progression e.g. Age-Related Macular Degeneration. Recently described two-photon vision may be applied to microperimetric devices. Many subjects with well-known disease history could be investigated with newly developed instrumentation that tests ability of human eye to perceive near infrared radiation. The main limitation of this new method is a very high cost of the femtosecond laser. Facing this problem, we try to replace the femtosecond laser with lower cost fiber-optic picosecond light source. To compare these two lasers, we constructed dedicated measurement system. We performed measurements of two-photon vision threshold on healthy subjects for two different light sources - sub picosecond Kerr mode-locking solid-state laser and fiber-based picosecond laser. Experiments were conducted for an open circle flickering stimulus with 0.5 deg. diameter, for retinal locations varying from 0 deg. to 5.8 deg., using 4-2-1 threshold strategy that is well-known from classical microperimetry. Values of obtained thresholds are only 5 times higher for the fiber laser than that obtained by using the femtosecond laser, while it was expected to be about over 16 times higher. This fact requires further investigations. Nevertheless, the idea of replacement of the latter laser by relatively cheap fiber-optic one in ophthalmic devices for two-photon vision studies seems to be potentially promising.
Human subjects can detect infrared light at wavelengths over 1000 nm perceived as visible of the corresponding half wavelength. This is due to a two-photon process and requires the use of pulsed light sources well focused within the retina. We have developed an experimental system to measure, for the first time, the visual resolution of the eye when is stimulated with infrared (1043 nm) and compared with visible light (543 nm). Scanner mirrors were used to project letters of different sizes onto the retina in both wavelengths. Subjects performed a visual test to determine the smallest letter size that was distinguishable for each wavelength for a range of defocus values. An additional optical path was used to record the retinal images of the spot after reflection in the retina and double-pass through the optical media. The best visual acuity was obtained at different defocus locations corresponding to the chromatic difference between green and infrared. Although, there was some individual variability, visual acuity was found to be similar both in visible and infrared. The use of two-photon infrared vision may have some potential applications for vision in those cases were the optical media is opaque to visible wavelengths while keeping some transparency in the infrared.
Light sensation relies on photoisomerization of chromophores in rod and cone photoreceptor cells. Spectral sensitivity of these photoreceptor cells in the retina is determined by the absorption spectra of their pigments which covers a range from 400 nm to above 700 nm. Regardless the mechanism leading to visual pigment isomerization, light sensation is triggered every time visual pigment molecules change their conformation. Thus, two-photon absorption (TPA) should produce the same result (visual sensation) as single photon absorption of light. This observation was positively verified and published by our group. During human psychophysics experiments, we found that humans can perceive light in the infrared (IR) range as colors that match half of the wavelength of the applied laser beam. Other experiments and theoretical research, such as mouse electrophysiology, biochemical studies of TPA in rhodopsin or molecular modeling studies, confirmed that visual sensation can be triggered by TPA. There are few publications describing human near infrared (NIR) perception and no formal proposals to use this phenomenon to improve ophthalmic diagnosis and monitor treatment. Here we report that the use of novel instrumentation revealed that the sensitivity threshold for NIR vision depends on age.
Scattering and fluorescence images provide complementary information about the health condition of the human eye, so getting them in a single measurement, using a single device may significantly improve a quality of diagnosis as it has been already demonstrated in Spectralis (Heidelberg Eng.) OCT instrument. There is still challenge to improve quality of fundus autofluorescence (FAF) images. The biggest obstacle in obtaining in vivo images of sufficient quality is very low fluorescence signal. For eye safety reasons, and because of patient comfort, using highpower fluorescence excitation is not an adequate solution to the low signal problem. In this contribution we show a new detection method in the retinal autofluorescence imaging, which may improve the sensitivity. We used a fast modulated (up to 500 MHz) diode laser of wavelength 473 nm and detected fluorescence in the spectral range 500-680 nm by photomultiplier and lock-in amplifier. Average power of the collimated blue beam on the cornea used for FAF measurements was set to 50 μW, 10 μW, and even 4.5 μW.
Results of the recent application studies of laser-based techniques for conservation practice, carried out in the frames of
research and conservation projects are presented and discussed. The Gotland Sandstone Project covers laser cleaning,
spectroscopic techniques of the process monitoring and materials analysis. The post-processing effect is investigated by
means of LIBS, colorimetry, SEM and XEDS in order to answer questions regarding encrustation removal, discoloration
and alteration due to environmental pollution and laser interaction, respectively. The cleaning is monitored acoustically.
Slight differences in stone yellowing are revealed when applying laser in air or pure N2 environment which is ascribed to
material oxidation and partial combustion of surface remnants due to presence of O2 contributing to the final effect of
laser cleaning. Results of microscopic observation are supported by the presence of elements S, Al, Ca, and C which
prevail in crust. In case of restoration of the historical documents on paper the ablative cleaning and the nearly nondestructive
identification and composition analysis of surface layers such as contaminants, substrate and pigments are
performed. Spectra obtained by means of the LIPS technique for historical, originally contaminated and also artificially
soiled model samples reveal reach structures. The emission lines of Ca, Na, K, Al and Fe are ascribed to contaminants
because of intensities decreasing with prolonged surface irradiation during laser cleaning. Bands of Ti and Ba correspond
to white pigments (TiO2 and BaSO4) in the paper and are confirmed by the Raman spectra as well. For identification of
historical pigments the reference measurements on model substrates are performed, too.
Techniques of laser emission spectroscopy such as LIPS and LIF are applied to identify pastels and pigments composition for the use in conservation of historical documents. The question of data reliable for pigment identification by these techniques is considered. For model samples made of the cotton paper of chemical composition corresponding to the historical ones, and coated with pastels of different colors the LIPS and LIF spectra are recorded. Samples are excited by the pulsed Nd:YAG laser operating at 355 or 266 nm. The reference Raman spectra are collected, too. Bands characteristic for the blue pigments: PB15, PB29; violet one PV16, and yellow PY 184 are identified exclusively by LIPS in agreement with literature. Also additives such as the barium white found in the Scarlet pastel, and ultramarine (Na8-10[Al6Si6O24]S2-4) with titanium white (TiO2) in the Phthalo Blue are identified, and confirmed by the Raman technique as well. The pigments anthraquinone (PR 168), isoindolinone (PY 110) and monoazo (PY 74) are not revealed. In the LIF spectra only a broad band centered at 612 nm and corresponding to anthraquinone (red pastel) can be clearly assigned.
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