Cheryl Dawn DiCarlo

Comparison of optical coherence tomography imaging of cataracts with histopathology

This paper presents a comparison of in vivo optical coherence tomography (OCT) captured cataract images to subsequent histopathological examination of the lenticular opacities. OCT imaging was performed on anesthetized Rhesus monkeys, known as the delayed effects colony (DEC), with documented cataracts. These monkeys were exposed to several types of radiation during the mid and late 1960s. The radiation and age related cataracts in these animals were closely monitored using a unique grading system developed specifically for the DEC. In addition to this system, a modified version of a common cataract grading scheme for use in humans was applied. Of the original 18 monkeys imaged, lenses were collected at necropsy from seven of these animals, processed, and compared to OCT images. Results showed a direct correlation between the vertical OCT images and the cataractous lesions seen on corresponding histopathologic sections of the lenses. Based on the images obtained and their corresponding documented comparison to histopathology, OCT showed tremendous potential to aid identification and characterization of cataracts. There can be artifactual problems with the images related to movement and shadows produced by opacities. However, with the advent of increased speed in imaging and multiplanar imaging, these disadvantages may easily be overcome.

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.

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

Abstract• Background: In vivo retinal injury studies using ultrashort-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 μJ at 120 fs, compared with the minimum visible lesion (MVL) threshold of 0.43 μJ 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 μJ of energy.

In vivo imaging of the development of linear and nonlinear retinal laser effects using optical coherence tomography in correlation with histopathological findings

Optical Coherence Tomography (OCT) is a new, non-invasive diagnostic technique for high resolution optical 3D imaging, which was developed and applied to several different biological materials during the lasi; five years [1, 2, 3]. A unique application ofthis technique is the microscopical cross-sectional imaging ofpostenor structures ofthe eye which are not accessable with other high resolution techniques in-vivo neither with x-ray-imaging nor with high frequency ultrasound scanning. The superior spatial resolution on the order ofabout lOtm laterally and axially, the high signal-to-noise ratio ofmore than 100 db and the fast acquisition-time of one second for a two dimensional scan provides a technique for cross-sectional in-vivo-momtoring ofintraocular structures and therefore the possibility to study the time course of anatomical and pathological developments in the eye. The acute morphological changes of ocular structures and their biological healing response after shortterm impacts such as high-intensity laser exposures are ofparticular interest for the understanding of the mechanisms responsible for therapeutic laser-application in ophthal-mology as well as for laser injury to the eye. A correlation between cross-sectional OCT-images and structural findings using classical histopathological techniques facilitates a better interpretation ofthe characteristic patterns seen in OCTimages and defines the sensitivity ofthe OCT-technique to image morphological details. On the other hand preparational artefacts not avoidable in all histological procedures can be identified and analyzed by comparing histological micrographs with OCT-images of exactly the same structure. First results of an experimental study where retinal effects were produced in monkey eyes using laser pulses from 200 ms to 130 fs in duration are presented in this article. The applied energies from 5tJ to 50 mJ were able to induce the whole spectrum of biological effects possible in the eye, ranging from intraretinal microruptures to extensive thermal denaturation and massive preretinal hemorrhages [4, 5, 6].

Visible lesion thresholds from near-infrared pico- and nanosecond laser pulses in the primate eye

Minimum visible lesions (MVL) are reported for picosecond and nanosecond laser pulses at near-IR wavelengths in the primate eye, Macaca Mulatta. The 50 percent probability for damage (ED50) dosages are reported for the 24 hour for both MVL and fluorescein angiography visible lesion thresholds at the 95 percent confidence level. The thresholds decreased by as much as 48 percent between the 1- hour reading and were lower in all cases at 24 hours. MVL- (ED50) threshold doses were 19.1 uJ at 7 ns and 4.2 uJ and 4.6 uJ at 80 ps and 20 ps respectively. Our thresholds measured for the near-IR laser pulses were lower by a factor of 4 to 8 lower than previously reported values but almost an order in magnitude higher than visible MVL thresholds for similar pulsewidth in the visible wavelengths.

In vivo laser-induced breakdown in the rabbit eye

Threshold measurements for femtosecond laser pulsewidths have been made for retinal minimum visible lesions (MVLs) in Dutch Belted rabbit and rhesus monkey eyes. Laser-induced breakdown (LIB) thresholds in biological materials including vitreous, normal saline, tap water, and ultrapure water have been measured and reported using an artificial eye. We have recorded on video the first LIB causing bubble formation in any eye in vivo using albino rabbit eyes (New Zealand white) with 120- femtosecond (fs) pulses and pulse energies as low as 5 microjoules ((mu) J). These bubbles were clearly formed anterior to the retina within the vitreous humor and, with 60 (mu) J of energy, they lasted for several seconds before disappearing and leaving no apparent damage to the retina. We believe this to be true LIB because of the lack of pigmentation or melanin granules within the albino rabbit eye (thus no absorptive elements) and because of the extremely high peak powers within the 5-(mu) J, 120-fs laser pulse. These high peak powers produce self-focusing of the pulse within the vitreous. The bubble formation at the breakdown site acts as a limiting mechanism for energy transmission and may explain why high-energy femotsecond pulses at energies up to 100 (mu) J sometimes do not cause severe damage in the pigmented rabbit eye. This fact may also explain why it is so difficult to produce hemorrhagic lesions in either the rabbit or primate eye with 100-fs laser pulses.

Simultaneous Exposure Using 532 and 860 nm lasers for visible lesion thresholds in the rhesus retina.

The growth of commercially available, simultaneous multi-wavelength laser systems has increased the likelihood of possible ocular hazard. For example, many systems utilize frequency multiplying methods to produce combinations of visible, near-infrared, and ultraviolet wavelengths. Unfortunately, very little data exists to substantiate the current methods for estimating hazards from simultaneous lasing. To properly assess the retinal hazards from these wavelengths, the retinal effects of 10-s laser irradiation from 532 and 860 nm were determined in non-human primates for four different relative dosage combinations of these wavelengths. This pair of wavelengths represents the typical problem of a visible-wavelength laser combined with an in-band, infrared wavelength that is not as well focused at the retina—a situation difficult to address. To add confidence to the experimental results obtained, a theoretical thermodynamic model was developed to predict the minimal damage threshold for simultaneous wavelengths at 1 h post exposure. The new model calculations and the data obtained are compared with results from one currently accepted method of predicting relative exposure limits from multi-wavelength systems. In addition, the current ANSI-Z136-2000 standard was used to compute the combined MPEs for comparison with measured visible lesion thresholds. A total of 12 eyes were exposed using four different ratios of power levels (532/860 power rations) to determine the contribution to the damage levels from each wavelength. The experimental data were analyzed using probit analysis at both 1-h and 24-h post exposure to determine the minimum-visible-lesion (MVL) thresholds at ED50 values, and these thresholds at 24 h varied from 5.6 mW to 17 mW total intraocular power.

Histopathology of ultrashort-laser-pulse retinal damage

Recent studies of retinal damage due to ultrashort laser pulses have shown interesting behavior. Laser induced retinal damage for ultrashort (i.e. less than 1 ns) laser pulses is produced at lower energies than in the nanosecond to microsecond laser pulse regime and the energy required for hemorrhagic lesions is much greater times greater for the nanosecond regime. We investigated the tissue effects exhibited in histopathology of retinal tissues exposed to ultrashort laser pulses.

New noninvasive imaging technique for cataract evaluation in the rhesus monkey

We present the first in vivo study using Optical Coherence Tomography (OCT) as the imaging device for lenticular cataracts in the geriatric rhesus monkey. OCT is a non-invasive imaging technique that produces a 2D cross sectional image of intraocular tissue similar to ultrasound B scan. In OCT the images are formed by measuring optical reflections from the tissue. Eighteen geriatric subjects with documented lenticular opacities and one control subject were imaged. The OCT images produced are compared to current and previous clinical cataract grading exams and slit-lamp photography. Histopathology was collected on one subject and is compared to the OCT image. OCT provides information on nuclear, cortical and subcapsular opacities. The image formation is presented based on a color coded computer generated log reflective scale. The log reflective scale is converted to a qualitative grading system. Although movement and shadow artifact can occur, these are readily identifiable and can be differentiated from underlying lenticular abnormalities. OCT has great potential to assist in further characterization of cataracts.

Effect of light emitting diode (LED) therapy on the survival of photoreceptors following argon laser injury

Due to the increasing number of optic systems that military personnel are exposed, the development of countermeasures for laser eye injury is of significant concern. Recent reports in the literature suggest some benefit form the use of Light Emitting Diode (LED) therapy on the retina that received a toxic insult. The purpose of this study was to compare retinal cell survival and multifocal electroretinography (mfERG) in a laser retinal injury model following treatment with LED photoillumination. Control and LED array (670 nm) illuminated cynomolgus monkeys received macular Argon laser lesions (514 nm, 130 mW, 100 ms). LED array exposure was accomplished for 4 days for a total dose of 4 J/cm2 per day. Baseline and post-laser exposure mfERGs were performed on most of the subjects. Ocular tissues were collected from four animals at Day 4 poast laser exposure and from two animals at 4 months post laser exposure. The tissues were processed for plastic embedding. Retinal cell counts were performed on the lesion sections. Analysis of Variance (ANOVA) results yielded no significant difference in the sparing of photoreceptors, inner nuclear and ganglion cells between the control and LED illuminated subjects. Although pathology showed no significant support for diode therapy, our early mfERG observations previously reported suggested a more rapid functional recovery. Since there is still no uniform therapy for laser retinal injury, research is continuing to determine novel therapies that may provide retinal cell sparing and functional retinal return.