Pathology of macular lesions from subnanosecond pulses of visible laser energy.

PURPOSE: To demonstrate how current theories regarding ultrashort laser pulse effects may apply to ocular tissue, a prospective clinicopathologic study of macular lesions from ultrashort laser pulses compared the pathologic effects with the clinical and fluorescein angiographic appearance of the laser lesions. METHODS: Ninety-femtosecond, 3-picosecond, and 60-picosecond laser pulses, throughout a range of energies, were delivered to the retina of Macaca mulatta. Clinical examination and fluorescein angiography were performed at 1 hour in all eyes and 24 hours after exposure in selected eyes. Eyes were enucleated at 1 or 24 hours after lesion placement. The structure and extent of retinal lesions were scored for comparison with the clinical findings. RESULTS: Focal retinal pathologic appearance correlated well with a clinically visible lesion observed 24 hours after laser delivery. Retinal lesions were small foci of retinal pigment epithelium (RPE) and retinal disruption, without choriocapillaris involvement. Lesions that contained focal RPE vacuoles or lifting of the RPE also demonstrated leakage, in fluorescein angiographic studies. Suprathreshold laser delivery frequently caused focal columns of retinal injury and intraretinal hemorrhages from retinal vessel bleeding, with no rupture of choroidal blood vessels. CONCLUSIONS: The retinal response to ultrashort laser pulses at moderate energy followed a pattern of focal damage from laser-induced breakdown without significant thermal spread.

Retinal effects of ultrashort laser pulses in the rabbit eye.

PURPOSE: To evaluate the effects of ultrashort laser pulses from femtoseconds to nanoseconds on the retinas of live rabbit eyes and to determine the energy requirements for visible lesion development. METHODS: The retinal effects of laser exposures were examined for laser exposures with pulsewidths ranging from 4 ns to 90 fs, with visible wavelengths of 532 nm for durations > 5 ps and 580 nm for durations < 5 ps. The authors examined and scored all laser impact sites in the retina ophthalmoscopically--with fundus photography and with fluorescein angiography--to identify evidence of visible laser effects. RESULTS: The laser energy required for retinal minimal visible lesions was found to be slightly less for pulsewidths < 5 ps and varied from 5 microJ at 4 ns to 1.1 microJ at 90 fs for the 1-hour ophthalmoscopic reading. Lesions from higher energy pulses (7 to 120 microJ) were examined at all pulsewidths. For 90-fs high-energy pulse delivery, an increased intensity of retinal lesions and the development of several subretinal hemorrhages were demonstrated at peak energies of 30 microJ. Fluorescein angiography was found to be much more sensitive as an indicator of retinal damage for both femtosecond pulsewidths. CONCLUSIONS: The low energies required for visible lesion production in live rabbit eyes raise new questions surrounding ultrashort pulse propagation in ocular media, energy deposition at the retina, and mechanisms limiting retinal damage from ultrashort laser pulses.

Visible retinal lesions from ultrashort laser pulses in the primate eye.

PURPOSE: To evaluate the effects of ultrashort laser pulses of visible wavelengths on the retinas of rhesus monkey eyes and to perform threshold measurements for minimum visible lesions (MVLs) at pulsewidths from nanoseconds to femtoseconds. METHODS: Single laser pulses at visible wavelengths were placed within the macular area of live rhesus monkey eyes at varying pulse energies at five pulsewidths (4 ns, 60 ps, 3 ps, 600 fs, and 90 fs). The number of visible lesions was determined after 1 hour and 24 hours postexposure, and a probit analysis was performed for the dosage, causing 50% probability for damage (ED50) as well as the 95% fiducial intervals for ED50. Fluorescein angiography (FA) was performed, and hemorrhagic lesions were recorded as they became visible. RESULTS: The ED50 threshold doses at the 1-hour reading, calculated from the measured data, decreased from 1.5 microJ at 4 ns to 0.60 microJ at 600 fs, but it increased to 1.18 microJ at 90 fs. At the 24-hour reading, the ED50 calculated doses decreased from 0.90 microJ at 4 ns down to 0.26 microJ at 600 fs, but it increased to 0.43 microJ at 90 fs. Fluorescein angiography visible lesion ED50 values were all higher than MVL values, showing that FA was not as sensitive in determining damage levels. CONCLUSIONS: Laser pulses for pulsewidths between 4 ns and 90 fs are capable of producing visible lesions in monkey eyes with energies less than 1 microJ. Fluorescein angiography is not as sensitive in determining threshold levels as visually observing the retina through a fundus camera.

Thresholds for visible lesions in the primate eye produced by ultrashort near-infrared laser pulses.

PURPOSE: To evaluate the effects of near-infrared (near-IR) ultrashort laser pulses on the retinas of rhesus monkey eyes and to perform threshold measurements for minimum visible lesions (MVLs) at pulse widths ranging from nanoseconds to femtoseconds. METHODS: Near-infrared single laser pulses were placed within the macular area of live rhesus monkey eyes for five different pulse widths (7 nsec; 80, 20, and 1 psec; and 150 fsec). One visible wavelength of 530 nm at 100 fsec was also included in the study. Visible lesion thresholds (MVL-ED50) were determined 1 hour and 24 hours after exposure. Fluorescein angiography thresholds (FAVL-ED50) were also determined using a probit analysis of the dosage. Thresholds were calculated as that dosage causing a 50% probability for damage, and the fiducial limits were calculated at the 95% confidence level. RESULTS: For all pulse widths, the 24-hour MVL-ED50 was lower than the 1-hour MVL-ED50, and they both decreased with decreasing pulse width. Thresholds at the 1-hour reading decreased from 28.7 microJ at 7 nsec to 1.8 microJ at 150 fsec, whereas thresholds at 24 hours decreased from 19.1 microJ at 7 nsec to 1.0 microJ at 150 fsec. The doubled 1060-nm wavelength of the 530-nm threshold decreased from 0.36 to 0.16 microJ after 24 hours. FAVL-ED50s were much higher than MVL-ED50s, showing that FA was not as sensitive in determining damage levels. CONCLUSIONS: Laser pulse widths less than 1 nsec in the near-IR are capable of producing visible lesions in rhesus monkey eyes with pulse energies between 5 and 1 microJ. Also, the near-IR thresholds for these pulse widths are much higher than for the visible wavelengths. As with visible wavelengths, FA is not as sensitive in determining threshold levels as is visually observing the retina through a fundus camera.

Argon laser retinal lesions evaluated in vivo by optical coherence tomography.

PURPOSE: To assess the in vivo evolution of argon laser retinal lesions by correlating the cross-sectional structure from sequential optical coherence tomography with histopathologic sectioning. METHODS: Argon laser lesions were created in the retinas of Macaca mulatta and evaluated by cross-section optical coherence tomography, which was compared at selected time points with corresponding histopathology. RESULTS: Argon laser lesions induced an optical coherence tomography pattern of early outer retinal relative high reflectivity with subsequent surrounding relative low reflectivity that correlated well with histopathologic findings. The in vivo optical coherence tomography images of macular laser lesions clearly demonstrated differences in pathologic response by retinal layer over time. CONCLUSION: The novel sequential imaging of rapidly evolving macular lesions with optical coherence tomography provides new insight into the patterns of acute tissue response by cross-sectional layer. This sequential imaging technique will aid in our understanding of the rapid evolution of retinal pathology and response to treatment in the research and clinical setting.

Comparison of macular versus paramacular retinal sensitivity to femtosecond laser pulses.

Single 130 fs laser pulses in the near-IR (800 nm) were used to create ophthalmoscopically viewed minimum visible lesions (MVLs) within the macular and paramacular regions in rhesus monkey eyes. MVL thresholds at 1 and 24 h are reported as the 50% probability for damage (ED50) together with their fiducial limits at the 95% confidence level. These measured thresholds are compared with previously reported thresholds for near-IR and visible wavelengths for both macular and paramacular areas. Threshold doses were lower at the 24 h reading than at the 1 h reading for both retinal regions and the ED50s for the macula were slightly lower than for the paramacula. We measured the 24 h MVL ED50 thresholds to be 0.35 and 0.55 microJ for the macular and paramacular areas, respectively. The combined data for both areas yielded a threshold of 0.45 microJ.

Comparative study of laser damage threshold energies in the artificial retina.

Laser damage threshold energies produced from ultrashort (i.e., ⩽1 ns) laser pulses are investigated as a function of both pulse width and spot size for an artificial retina. A piece of film acts as the absorbing layer and is positioned at the focus of a variant on the Cain artificial eye [C. Cain, G. D. Noojin, D. X. Hammer, R. J. Thomas, and B. A. Rockwell, "Artificial eye for in vitro experiments of laser light interaction with aqueous media," J. Biomed. Opt.2, 88-94 (1997)]. Experiments were performed at the focal point and at two and ten Rayleigh ranges (RR) in front of the focus with the damage end point being the presence of a bubble imaged at the film plane. Pulse energy thresholds were determined for wavelengths of 1064, 580, and 532 nm with pulse durations ranging from the nanosecond (ns) to the femtosecond (fs) regime. For the at-focus data in the visible regime, the threshold dropped from 0.25 µJ for a 532 nm, 5 ns pulse to 0.11 µJ for a 580 nm, 100 fs pulse. The near-infrared (NIR) threshold changed from 5.5 µJ for a 5 ns pulse to 0.9 µJ for a 130 fs pulse at a distance two RR in front of the focus. The experiment was repeated using the same pulse widths and wavelengths, except the water path was removed to determine the impact of nonlinear self-focusing in water. A vertical microscope imaging system was employed in order to observe the threshold event. The NIR fluence threshold of 0.5 J/cm2 remained constant within an experimental uncertainty for all pulse widths, which corresponds to values in the literature [C. P. Lin and M. W. Kelly, "Ultrafast time-resolved imaging of stress transient and cavitation from short pulsed laser irradiated melanin particles," SPIE Laser-Tissue Interactions VI, Proc. SPIE2391, 294-299 (1995)]. The visible data also demonstrated a nearly constant fluence of 0.07 J/cm2. The disparity in thresholds between the two techniques arises from nonlinear optical phenomena related to propagation differences in the ocular fluid. © 1999 Society of Photo-Optical Instrumentation Engineers.

Ultrashort laser pulse bioeffects and safety.

Recent studies of retinal damage due to ultrashort laser pulses have shown that less energy is required for retinal damage for pulses shorter than 1 ns than that for longer pulses. It has also been shown that more energy is required for near-infrared (NIR) wavelengths than in the visible because the light focuses behind the retina, requiring more energy to produce a damaging fluence on the retina. We review the progress made in determining the trends in retinal damage from laser pulses of 1 ns to 100 fs in the visible and NIR wavelength regimes. We have determined the most likely damage mechanism(s) operative in this pulse width regime.

Spectrally resolved white-light interferometry for measurement of ocular dispersion.

Spectrally resolved white-light interferometry was used to measure the wavelength dependence of refractive index (i.e., dispersion) for various ocular components. Verification of the technique's efficacy was substantiated by accurate measurement of the dispersive properties of water and fused silica, which have both been well-characterized in the past by single-wavelength measurement of the refractive index. The dispersion of bovine and rabbit aqueous and vitreous humors was measured from 400 to 1100 nm. In addition, the dispersion was measured from 400 to 700 nm for aqueous and vitreous humors extracted from goat and rhesus monkey eyes. An unsuccessful attempt was also made to use the technique for dispersion measurement of bovine cornea and lens. The principles of white-light interferometry, including image analysis, measurement accuracy, and limitations of the technique, are discussed. In addition, alternate techniques and previous measurements of ocular dispersion are reviewed.

Intraocular laser surgical probe for membrane disruption by laser-induced breakdown.

A fiber probe has been designed as a surgical aid to cut intraocular membranes with laser-induced breakdown as the mechanism. The design of the intraocular laser surgical probe is discussed. A preliminary retinal damage distance has been calculated with breakdown threshold, spot size, and shielding measurements. Collateral mechanical-damage effects caused by shock wave and cavitation are discussed.

Shielding properties of laser-induced breakdown in water for pulse durations from 5 ns to 125 fs.

The shielding effectiveness of laser-induced breakdown from focused, visible laser pulses from 5 ns to 125 fs is determined from measurements of transmission of energy through the focal volume. The shielding efficiency decreases as a function of pulse duration from 5 ns to 300 fs and increases from 300 fs to 125 fs. The results are compared with past studies at similar pulse durations. The results of the measurements support laser-induced breakdown models and may lead to an optimization of laser-induced breakdown in ophthalmic surgery by reduction of collateral effects.

Nonlinear refraction in vitreous humor.

We extend the application of the z-scan technique to determine the nonlinear refractive index (n(2)) for human and rabbit vitreous humor, water, and physiological saline. In these measurements there were nonlinear contributions to the measured signal from the aqueous samples and the quartz cell that held the sample. Measurements were made with 60-ps pulses at 532 nm. To our knowledge, this is the first measurement of the nonlinear refractive properties of biological material.

Retinal damage and laser-induced breakdown produced by ultrashort-pulse lasers.

BACKGROUND: In vivo retinal injury studies using ultra-short-pulse lasers at visible wavelengths for both rabbit and primate eyes have shown that the degree of injury to the retina is not proportional to the pulse energy, especially at suprathreshold levels. In this paper we present results of calculations and measurements for laser-induced breakdown (LIB), bubble generation, and self-focusing within the eye. METHODS: We recorded on video and measured the first in vivo LIB and bubble generation thresholds within the vitreous in rabbit and primate eyes, using external optics and femtosecond pulses. These thresholds were then compared with calculations from our LIB model, and calculations were made for self-focusing effects within the vitreous for the high peak power pulses. RESULTS: Results of our nonlinear modeling and calculations for self-focusing and LIB within the eye were compared with experimental results. The LIB ED50 bubble threshold for the monkey eye was measured and found to be 0.56 microJ at 120 fs, compared with the minimum visible lesion (MVL) threshold of 0.43 microJ at 90 fs. Self-focusing effects were found to be possible for pulsewidths below 1 ps and are probably a contributing factor in femtosecond-pulse LIB in the eye. CONCLUSIONS: Based on our measurements for the MVL thresholds and LIB bubble generation thresholds in the monkey eye, we conclude that in the femtosecond pulsewidth regime for visible laser pulses, LIB and self-focusing are contributing factors in the lesion thresholds measured. Our results may also explain why it is so difficult to produce hemorrhagic lesions in either the rabbit or primate eye with visible 100-fs laser pulses even at 100 microJ of energy.