Evaluation of human sclera after femtosecond laser ablation using two photon and confocal microscopy.

Glaucoma is the second-leading cause of blindness worldwide and is often associated with elevated intraocular pressure (IOP). Partial thickness intrascleral channels can be created with a femtosecond laser operating at a wavelength of 1700 nm. Such channels have the potential to increase outflow facility and reduce elevated IOP. Analysis of the dimensions and location of these channels is important in understanding their effects. We describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in human cadaver eyes. High-resolution images, hundreds of microns deep in the sclera, were obtained to allow determination of the shape and dimension of such channels. This demonstrates that concept of integrating femtosecond laser surgery, and two-photon and confocal imaging has the future potential for image-guided high-precision surgery in transparent and translucent tissue.

Finite element model of the temperature increase in excised porcine cadaver iris during direct illumination by femtosecond laser pulses.

In order to model the thermal effect of laser exposure of the iris during laser corneal surgery, we simulated the temperature increase in porcine cadaver iris. The simulation data for the 60 kHz FS60 Laser showed that the temperature increased up to 1.23°C and 2.45°C (at laser pulse energy 1 and 2 [micro sign]J, respectively) by the 24 second procedure time. Calculated temperature profiles show good agreement with data obtained from ex vivo experiments using porcine cadaver iris. Simulation results of different types of femtosecond lasers indicate that the Laser in situ keratomileusis procedure does not present a safety hazard to the iris.

Temperature increase in porcine cadaver iris during direct illumination by femtosecond laser pulses.

PURPOSE: To measure the temperature rise in porcine cadaver iris during direct illumination by the femtosecond laser as a model for laser exposure of the iris during femtosecond laser corneal surgery. SETTING: Department of Ophthalmology, University of California-Irvine, Irvine, California, USA. DESIGN: Experimental study. METHODS: The temperature increase induced by a 60 kHz commercial femtosecond laser in porcine cadaver iris was measured in situ using an infrared thermal imaging camera at pulse energy levels ranging from 1 to 2 µJ (corresponding approximately to surgical energies of 2 to 4 µJ per laser pulse). RESULTS: Temperature increases up to 2.3 °C (corresponding to 2 µJ and 24-second illumination) were observed in the porcine cadaver iris with little variation in temperature profiles between specimens for the same laser energy illumination. CONCLUSIONS: The 60 kHz commercial femtosecond laser operating with pulse energies at approximately the lower limit of the range evaluated in this study would be expected to result in a 1.2 °C temperature increase and therefore does not present a safety hazard to the iris.

Temperature increase in human cadaver retina during direct illumination by femtosecond laser pulses.

PURPOSE: Femtosecond lasers have been approved by the US Food and Drug Administration for ophthalmic surgery, including use in creating corneal flaps in LASIK surgery. During normal operation, approximately 50% to 60% of laser energy may pass beyond the cornea, with potential effects on the retina. As a model for retinal laser exposure during femtosecond corneal surgery, we measured the temperature rise in human cadaver retinas during direct illumination by the laser. METHODS: The temperature increase induced by a 150-kHz iFS Advanced Femtosecond Laser (Abbott Medical Optics) in human cadaver retinas was measured in situ using an infrared thermal imaging camera. To model the geometry of the eye during the surgery, an approximate 11x11-mm excised section of human cadaver retina was placed 17 mm behind the focus of the laser beam. The temperature field was observed in 10 cadaver retina samples at energy levels ranging from 0.4 to 1.6 microJ (corresponding approximately to surgical energies of 0.8 to 3.2 microJ per pulse). RESULTS: Maximal temperature increases up to 1.15 degrees C (corresponding to 3.2 microJ and 52-second illumination) were observed in the cadaver retina sections with little variation in temperature profiles between specimens for the same laser energy illumination. CONCLUSIONS: The commercial iFS Advanced Femtosecond Laser operating with pulse energies at approximately the lower limit of the range evaluated in this study would be expected to result in a 0.2 degrees C temperature increase and do not therefore present a safety hazard to the retina.

Imaging pulsatile retinal blood flow in human eye.

A functional Fourier domain optical coherence tomography instrument offering spectral Doppler imaging of in vivo pulsatile human retinal blood flow was constructed. An improved phase-resolved algorithm was developed to correct bulk motion artifacts. Spectral Doppler imaging provides complementary temporal flow information to the spatially distributed flow information of the color Doppler image by providing direct visualization of the Doppler spectrum of the flow whose pattern can be further quantified with various velocity envelope curves and their corresponding flow indices. The coefficient of repeatability on resistance index measurement was assessed by analyzing 14 measurements on two vessels within two normal subjects.

Corneal response to femtosecond laser photodisruption in the rabbit.

In this report we evaluated the effect of femtosecond laser energy on the development of corneal haze and keratocyte activation in rabbits following intra-stromal photodisruption to create LASIK flaps using a modified commercial femtosecond surgical laser. Three groups of flap parameters were studied: 1.5 microJ/pulse with 10 microm spot separation and complete side cut (Group 1); 3.5 microJ/pulse with 14 microm spot separation and complete side cut (Group 2); 3.5 microJ/pulse with 14 microm spot separation and partial (50 microm) side cut (Group 3). All flaps were left attached without lifting to avoid epithelial contamination. Rabbits were then evaluated pre- and post-operatively by quantitative in vivo and ex vivo confocal microscopy. The achieved flap thickness 1 week after surgery averaged 88.9+/-12.8, 90.8+/-6.9 and 86.5+/-6.8 microm for Groups 1-3 respectively (p=NS). Interface thickness was significantly greater (p<0.05) in the higher energy groups averaging 40.0+/-11.2 and 37.7+/-5.7 microm for Groups 2-3 compared to 28.6+/-4.5 microm for Group 1. Corneal haze was barely detectible and not significantly different between groups, although haze was detected in the region of the side-cuts in Groups 1 and 2. No clinically significant changes in stromal or epithelial thickness were noted. Laser confocal microscopy showed the presence of small diameter cells within the flap interface that resided within disrupted regions of the corneal collagen lamellae. Keratocyte activation was only detected in regions of the 100% side cut and not over the flap interface. In conclusion, the results of this study indicate that photodisruption of the corneal stroma alone without flap elevation regardless of laser energy does not induce significant corneal haze in the rabbit. However, a thicker stromal interface was seen with the higher energy suggesting greater stromal damage.

Characterization of submicrojoule femtosecond laser corneal tissue dissection.

PURPOSE: To document the acute morphologic features of laser of situ keratomileusis (LASIK) flaps created using an IntraLase femtosecond laser (IntraLase, Inc.) with a 60 kHz engine. SETTING: Laser suite in a clinical practice. METHODS: A LASIK flap was created in 4 human eye-bank eyes using the 60 kHz IntraLase femtosecond laser with the following settings: 110 microm flap thickness, 9.0 mm flap diameter, 60-degree hinge length, 65-degree side cut, 0.4 muJ or 0.7 muJ raster energy, 7 microm x 7 microm or 9 microm x 9 microm spot/line separation, and 1 muJ side-cut energy. Immediately after the laser pass and without the flap being lifted, the globes were placed in fixative and subsequently processed for light and transmission electron microscopy. RESULTS: All 4 procedures were completed without complications or the appearance of an opaque bubble layer. The flaps were of uniform thickness and equaled the attempted thickness. Some areas had a complete dissection; other areas had scattered, incomplete tissue bridges. The adjacent corneal stroma and keratocytes were uninjured. When the epithelium was removed, the stromal component of the flap was measured as the attempted thickness; when the epithelium was present, the total flap thickness approximated the attempted flap thickness. CONCLUSIONS: Laser in situ keratomileusis flaps were safely created using raster energies and laser spot separations below those being used clinically. This technique may allow creation of flaps that are reproducibly thinner than those currently being performed and thus confer the benefits of surface ablation and LASIK.

Accuracy of digital images for assessing diabetic retinopathy.

BACKGROUND: To determine the accuracy of diabetic retinopathy status assessments with and without pupil dilation using digital fundus photographs acquired by a clinic staff person and interpreted remotely by ophthalmologists. METHOD: Using early treatment diabetic retinopathy study (EDTRS) grading criteria, diabetic retinopathy status assessments were made by an experienced (nonphysician) retinal grader (NPG) based on seven standard field 35-mm stereoscopic slides acquired by an experienced ophthalmic photographer. These assessments were compared with those of the same eyes made by two ophthalmologists and the NPG using digital photographs acquired by a clinic staff person using a high-resolution (800 x 600) digital color camera system. RESULTS: Based on 35-mm slides, 38% of 244 diabetic patients had ETDRS > or =35 in at least one eye and 5% had vision-threatening diabetic retinopathy (ETDRS > or =53 or macular edema). The proportion of ungradable images was significantly greater for nonmydriatic than mydriatic assessments (30% versus 10% ungradable as determined by the NPG). For ETDRS level > or =35, specificity ranged from moderate to high (0.70 to 0.96) for the three graders, while sensitivity was poor to moderate (0.38 to 0.71), and the area under the receiver-operating characteristic curves was less than satisfactory (0.67 to 0.71). CONCLUSIONS: The low sensitivity of the digital assessments indicates a significant proportion of patients in need of referral would not have been referred. These findings suggest that implementation of a simplified screening system using nonphotographer clinic staff acquiring nonmydriatic images, with interpretation by an ophthalmologist, should take place with an understanding of potential limitations.

Finite-element modeling of posterior lamellar keratoplasty: construction of theoretical nomograms for induced refractive errors.

PURPOSE: To estimate the theoretical corneal refractive error induced by mechanical weakening effects from posterior lamellar keratoplasty (PLKP) in the human cornea. METHODS: The refractive effects of PLKP are simulated by finite-element modeling (FEM) as a mathematical function of the thickness of the excised posterior lamellar corneal button, with a nonlinear formulation of stress-strain relation for the corneal material. A theoretical nomogram was developed to correlate the refractive changes to button thickness. RESULTS: The predicted refractive change after PLKP is less than 1 dpt for a 170-microm thickness posterior corneal button over a broad range of Young's modulus. Thicker buttons result in greater surgically induced refractive errors. CONCLUSIONS: According to FEM analysis, the excision of a posterior lamellar button of less than 170 microm thickness produces a minimal predicted refractive change (< 1 dpt) in the cornea after PLKP.

Top hat wound configuration for penetrating keratoplasty using the femtosecond laser: a laboratory model.

PURPOSE: To evaluate the mechanical stability and induced astigmatism of a modified multiplanar "top hat" wound configuration for full-thickness penetrating keratoplasty (PK) using the femtosecond laser as compared with PK in a laboratory model. METHODS: Eight human corneoscleral rims were mounted on an artificial anterior chamber. Four samples were assigned to the traditional PK group. Four samples underwent full-thickness keratoplasty with the femtosecond laser: a 9.0-mm cylindrical cut was made from the anterior chamber into the stroma, followed by a ring-shaped (outer diameter 9.0 mm, inner diameter 7.0 mm) horizontal lamellar resection at two-thirds corneal depth and a 7.0-mm cylindrical cut from the lamellae to the corneal surface. Mechanical stability was evaluated after placement of the cardinal sutures and the running sutures. RESULTS: In the "top hat" PK group, wound leakage occurred at 19 +/- 3.36 mm Hg after placement of the cardinal sutures and at 86.25 +/- 9.74 mm Hg after placement of the running sutures. In the traditional PK group, leakage occurred at 0 +/- 0 mm Hg and 76.25 +/- 20.98 mm Hg after placement of the cardinal sutures and running sutures, respectively. Both techniques induced steepening of the corneal curvature postop. The modified wound group showed a mean change in average K of 3.43 +/- 3.62 D, whereas the traditional PK group showed a mean change in average K of 3.21 +/- 6.67 D. CONCLUSION: The femtosecond laser-produced "top hat" wound configuration for PK was found to be more mechanically stable than that produced by the traditional method.

Femtosecond laser-assisted intracorneal keratoprosthesis implantation: a laboratory model.

PURPOSE: To demonstrate femtosecond laser-assisted intracorneal keratoprosthesis implantation and determine the mechanical stability as a function of intraocular pressure. METHODS: Eight human corneoscleral rims were mounted on an artificial anterior chamber. The femtosecond laser microkeratome was used to create a 2.5-mm diameter posterior corneal cap. A 7.2-mm-diameter lamellar stromal pocket was then created at mid-corneal depth. Finally, a 6-mm arc opening to the corneal surface was created at the periphery of the lamellar cut. The posterior lenticule was removed using corneal forceps and a 7.0-mm biopolymer keratoprosthesis was inserted into the stromal pocket. The surface wound was sealed using two 10-0 nylon sutures. A 3.0-mm anterior corneal opening was trephined to expose the keratoprosthesis. Intrachamber pressure was raised until wound leak was observed. RESULTS: Seven of the 8 implants withstood pressures of at least 135 mm Hg without implant extrusion. CONCLUSION: Femtosecond laser corneal dissection provides an alternative to more challenging manual dissection methods for keratoprosthesis implantation. Use of the femtosecond laser microkeratome will further refine keratoprosthesis surgical technique and may allow rapid and easy execution of the surgery.

Femtosecond laser posterior lamellar keratoplasty: a laboratory model.

PURPOSE: To evaluate feasibility of femtosecond laser application in posterior lamellar keratoplasty. METHODS: To evaluate the laser's effectiveness through opaque corneas, anterior corneal caps were resected from opaque corneas induced with 80% acetone solution. To evaluate the femtosecond laser posterior lamellar keratoplasty surgical procedure, human corneoscleral rims were mounted on an artificial anterior chamber. After corneal pachymetry, the femtosecond laser was used to create a 6-mm-diameter, 200-microm-thick endostromal lenticule. Access to the lenticule was provided by a small perilimbal surface opening, also created by the laser. The lenticule was removed using a pair of corneal forceps. A donor lenticule of similar dimensions was created, its endothelial surface coated with viscoelastic, inserted, and positioned on the recipient bed. Two sutures were placed to seal the small surface opening. RESULTS: The femtosecond laser produced an effective and smooth dissection through opaque corneas even at deeper settings. Graft transplantation was fairly simple and effective. CONCLUSION: Femtosecond laser posterior lamellar keratoplasty is a procedure that may provide an alternative to penetrating keratoplasty or the technically challenging manual posterior lamellar keratoplasty.

Subsurface photodisruption in human sclera: wavelength dependence.

BACKGROUND AND OBJECTIVE: Approximately 105 million people worldwide have glaucoma, and approximately 5 million are blind from its complications. Current surgical techniques often fail because of scarring of the conjunctival tissue, Tenon's tissue, or both. Femtosecond lasers can create highly precise incisions beneath the surface of a tissue, as previously demonstrated in the transparent cornea. Because the sclera is a highly scattering subsurface, photodisruption has not been previously possible. MATERIALS AND METHODS: To overcome scattering, a laser operating at 1,700 nm was used to make subsurface cuts in human sclera in vitro via photodisruption. RESULTS: Sub-10-microm width incisions were created beneath the surface without collateral tissue effects, something not possible with shorter wavelengths used to date in corneal applications with the femtosecond laser. CONCLUSION: Completely subsurface photodisruptions can be accomplished in human sclera in vitro. In vivo studies are required to evaluate the potential use of this technology for scleral applications.