Imaging ex vivo healthy and pathological human brain tissue with ultra-high-resolution optical coherence tomography.

The ability of ultra-high-resolution optical coherence tomography (UHR OCT) to discriminate between healthy and pathological human brain tissue is examined by imaging ex vivo tissue morphology of various brain biopsies. Micrometer-scale OCT resolution (0.9x2 microm, axialxlateral) is achieved in biological tissue by interfacing a state-of-the-art Ti:Al2O3 laser (lambda(c)=800 nm, delta lambda=260 nm, and P(out)=120 mW exfiber) to a free-space OCT system utilizing dynamic focusing. UHR OCT images are acquired from both healthy brain tissue and various types of brain tumors including fibrous, athypical, and transitional meningioma and ganglioglioma. A comparison of the tomograms with standard hematoxylin and eosin (H&E) stained histological sections of the imaged biopsies demonstrates the ability of UHR OCT to visualize and identify morphological features such as microcalcifications (>20 microm), enlarged nuclei of tumor cells (approximately 8 to 15 microm), small cysts, and blood vessels, which are characteristic of neuropathologies and normally absent in healthy brain tissue.

Adaptive-optics ultrahigh-resolution optical coherence tomography.

Merging of ultrahigh-resolution optical coherence tomography (UHR OCT) and adaptive optics (AO), resulting in high axial (3 microm) and improved transverse resolution (5-10 microm) is demonstrated for the first time to our knowledge in in vivo retinal imaging. A compact (300 mm x 300 mm) closed-loop AO system, based on a real-time Hartmann-Shack wave-front sensor operating at 30 Hz and a 37-actuator membrane deformable mirror, is interfaced to an UHR OCT system, based on a commercial OCT instrument, employing a compact Ti:sapphire laser with 130-nm bandwidth. Closed-loop correction of both ocular and system aberrations results in a residual uncorrected wave-front rms of 0.1 microm for a 3.68-mm pupil diameter. When this level of correction is achieved, OCT images are obtained under a static mirror configuration. By use of AO, an improvement of the transverse resolution of two to three times, compared with UHR OCT systems used so far, is obtained. A significant signal-to-noise ratio improvement of up to 9 dB in corrected compared with uncorrected OCT tomograms is also achieved.

Full range complex spectral optical coherence tomography technique in eye imaging.

We demonstrate a new implementation of complex spectral optical coherence tomography (OCT) in biomedical imaging. By reconstruction of both amplitude and phase we are able to use the negative and positive optical path differences to get images of objects of considerable thickness. An accompanying reduction of coherent noise improves the quality of the images. The property of the complex spectral OCT that permits the measurement range to be increased and permits the simultaneous use of phase and amplitude in spectral systems was not described previously. To show the potential of this technique we measured an anterior chamber of a porcine eye in vitro.

Submicrometer axial resolution optical coherence tomography.

Optical coherence tomography (OCT) with unprecedented submicrometer axial resolution achieved by use of a photonic crystal fiber in combination with a compact sub-10-fs Ti:sapphire laser (Femtolasers Produktions) is demonstrated for what the authors believe is the first time. The emission spectrum ranges from 550 to 950 nm (lambda(c)=725 nm , P(out)=27 mW) , resulting in a free-space axial OCT resolution of ~0.75 mum , corresponding to ~0.5 mum in biological tissue. Submicrometer-resolution OCT is demonstrated in vitro on human colorectal adenocarcinoma cells HT-29. This novel light source has great potential for development of spectroscopic OCT because its spectrum covers the absorption bands of several biological chromophores.

Improved prediction of intraocular lens power using partial coherence interferometry.

PURPOSE: To evaluate the feasibility of using a new optical biometry technique, dual-beam partial coherence interferometry (PCI), to improve intraocular lens (IOL) power prediction in cataract surgery. SETTING: Department of Ophthalmology, Vienna General Hospital, and Institute of Medical Physics, University of Vienna, Vienna, Austria. METHODS: Preoperative axial length (AL) data obtained with PCI biometry and applanation ultrasound (US) biometry in 77 eyes of 51 patients was applied to 4 commonly used IOL power formulas. The refractive outcome and the mean absolute error (MAE) were calculated for each formula using both biometry methods. A linear multiple-regression model based on preoperative PCI biometry data was derived to predict the postoperative anterior chamber depth (ACD). The predictive power of this regression model was assessed by adding the predicted ACD to the SRK/T formula. Predicted residuals were calculated to evaluate the feasibility and stability of this modified IOL power formula. RESULTS: Using PCI instead of US biometry significantly improved the refractive outcome with all 4 IOL power formulas. The Holladay I and SRK/T formulas yielded an MAE of 0.44 diopter (D) using PCI AL data and 0.56 D and 0.57 D, respectively, using US biometry data. The SRK/T formula combined with the PCI regression model for postoperative ACD prediction performed slightly better (MAE 0.42 D) than the conventional SRK/T formula alone. Predicted residuals revealed an MAE of 0.46 D, proving the predictive performance of the new formula. CONCLUSIONS: Partial coherence interferometry biometry applied to several widely used IOL power formulas yielded significantly better IOL power prediction and therefore refractive outcome in cataract surgery than US biometry. Further improvement can be achieved by applying PCI to a modified SRK/T formula that predicts the postoperative ACD using PCI biometry data.

Quantitative differential phase measurement and imaging in transparent and turbid media by optical coherence tomography.

Differential phase-contrast optical coherence tomography allows one to measure the path-length differences of two transversally separated beams in the nanometer range. We calculate these path-length differences from the phase functions of the interferometric signals. Pure phase objects consisting of chromium layers containing steps of approximately 100-200-nm height were imaged. Phase differences can be measured with a precision of +/-2 degrees , corresponding to a path-difference resolution of 2-3 nm. To investigate the influence of scattering, we imaged the phase objects through scattering layers with increasing scattering coefficients. The limit of phase imaging through these layers was at approximately 8-9 mean free path lengths thick (single pass).

Differential phase measurements in low-coherence interferometry without 2pi ambiguity.

Quantitative phase measurements by low-coherence interferometry and optical coherence tomography are restricted by the well-known 2pi ambiguity to path-length differences smaller than lambda/2 . We present a method that overcomes this ambiguity. Introducing a slight dispersion imbalance between reference and sample arms of the interferometer causes the short and long wavelengths of the source spectrum to separate within the interferometric signal. This causes the phase slope to vary within the signal. The phase-difference function between two adjacent sample beam components is calculated by subtraction of their phase functions obtained from phase-sensitive interferometric signal recording. Because of the dispersive effect, the phase difference varies across the interferometric signal. The slope of that phase difference is proportional to the optical path difference, without 2pi ambiguity.

Resolution-improved dual-beam and standard optical coherence tomography: a comparison.

BACKGROUND: The purpose of the study was to demonstrate the improved axial resolution and longitudinal stability of dual-beam optical coherence tomography (OCT) in comparison to conventional OCT setups used in commercially available OCT instruments. METHODS: The conventional OCT technique is based on an interferometric setup that is rather sensitive to axial eye motions. We have developed a special dual-beam OCT technique which eliminates the influence of axial eye motions. This is achieved by using the anterior corneal surface as the reference surface for the interferometric ranging. To improve the signal quality, the different wavefront curvatures of beams reflected at cornea and retina are matched by a diffractive optical element. To improve the axial resolution, a broadband synthesized light source with an effective bandwidth of 50 nm is used, and the group dispersion of the ocular media is compensated. Tomographic images were recorded in the fovea and the optic nerve head of healthy volunteers. For comparison purposes, approximately the same locations in the same eyes were imaged by a commercially available OCT instrument. RESULTS: Compared to the standard OCT technique, the dual-beam OCT images show considerably improved axial resolution. Especially in tomograms recorded at the fovea, dual-beam OCT resolves microstructural details that are not visible in the standard OCT images. Furthermore, the axial stability of dual-beam OCT enables the recording of exact geometrical contours of fundus layers. CONCLUSIONS: Dual-beam OCT is able to provide structural information on the ocular fundus that is not obtained with standard OCT. The long recording times of our instrument limit the transverse resolution to 100-150 microm at present.

Polarization-sensitive optical coherence tomography of dental structures.

Optical coherence tomography (OCT) has been developed during the last 10 years as a new noninvasive imaging tool and has been applied to diagnose different ocular and skin diseases. This technique has been modified for cross-sectional imaging of dental structures. In this first preliminary study the technique was applied to obtain tomographic images of extracted sound and decayed human teeth in order to evaluate its possible diagnostic potential for dental applications. Classical OCT images based on reflectivity measurements and phase retardation images using polarization-sensitive OCT were recorded. It was demonstrated that polarization-sensitive OCT can provide additional information which is probably related to the mineralization status and/or the scattering properties of the dental material. One of the attractive features of OCT is that it uses near-infrared light instead of ionizing radiation. Furthermore, high transversal and depth resolution on the order of 10 microm can be obtained. Present limitations, e.g. the limited penetration depth, and possible solutions are discussed.

Spectral measurement of absorption by spectroscopic frequency-domain optical coherence tomography.

A new method of measurement that essentially combines Fourier-domain optical coherence tomography with spectroscopy is introduced. By use of a windowed Fourier transform it is possible to obtain, in addition to the object structure, spectroscopic information such as the absorption properties of materials. The feasibility of this new method for performing depth-resolved spectroscopy is demonstrated with a glass filter plate. The results are compared with theoretically calculated spectra by use of the well-known spectral characteristics of the light source and the filter plate.

Changes in intraocular lens position after neodymium: YAG capsulotomy.

PURPOSE: To quantify changes in intraocular lens (IOL) position caused by neodymium: YAG (Nd:YAG) capsulotomy with 3 IOL styles. SETTING: Department of Ophthalmology, University of Vienna, Austria. METHODS: In a prospective study, anterior chamber depth (ACD) was measured by dualbeam partial coherence interferometry (PCI) in 32 pseudophakic eyes of 32 patients with posterior capsule opacification before and immediately after planned capsulotomy under mydriasis. Patients were divided into 3 groups with the following IOL styles: 1-piece poly(methyl methacrylate) (PMMA), 3-piece foldable, and plate haptic. RESULTS: The capsulotomy induced a backward IOL movement in all 32 eyes (mean 25 microns; range 9 to 55 microns). It was more pronounced in eyes with plate-haptic IOLs than in those with the other styles. Precision of ACD measurement by PCI was 4 microns. Changes in ACD correlated significantly with capsulotomy size but not with preoperative lens-capsule distance. CONCLUSION: Capsulotomy caused a backward movement of the IOL, which was more pronounced with plate-haptic IOLs than with 1-piece PMMA and 3-piece foldable IOLs. Since the magnitude of IOL movement in this study population was small, a hyperopic shift in refraction after capsulotomy will usually be small and not clinically relevant.

Differential phase contrast in optical coherence tomography.

We report on a modification of optical coherence tomography (OCT) that allows one to measure small phase differences between beams traversing adjacent areas of a specimen. The sample beam of a polarization-sensitive low-coherence interferometer is split by a Wollaston prism into two components that traverse the object along closely spaced paths. After reflection at the various sample surfaces, the beams are recombined at the Wollaston prism. Any phase difference encountered between the two beams is converted into a change of polarization state of the recombined beam. This change is measured, and the resulting signals are converted to differential phase-contrast OCT images. The first images obtained from simple test objects allowed us to determine path-difference gradients with a resolution of the order of 5 x 10(-5) .

Dispersion effects in partial coherence interferometry: implications for intraocular ranging.

In nondispersive media, the minimum distance that can be resolved by partial coherence interferometry (PCI) and optical coherence tomography (OCT) is inversely proportional to the source spectral bandwidth. Dispersion tends to increase the signal width and to degrade the resolution. We analyze the situation for PCI ranging and OCT imaging of ocular structures. It can be shown that for each ocular segment an optimum source bandwidth yielding optimum resolution exists. If the resolution is to be improved beyond this point, the group dispersion of the ocular media has to be compensated. With the use of a dispersion compensating element, and employing a broadband superluminescent diode, we demonstrate a resolution of 5 µm in the retina of both a model eye and a human eye in vivo. This is an improvement by a factor of 2-3 as compared to currently used instruments. © 1999 Society of Photo-Optical Instrumentation Engineers.

Partial coherence interferometry: a novel approach to biometry in cataract surgery.

PURPOSE: To compare biometry performed by an enhanced version of dual beam partial coherence interferometry and applanation ultrasound in a prospective study of 85 cataract eyes to improve refractive outcome of cataract surgery due to a more accurate calculation of intraocular lens power. METHODS: The SRK II formula using ultrasound biometry data was employed. Three months after surgery, partial coherence interferometry biometry was repeated and refractive outcome was determined. Preoperative partial coherence interferometry biometry data were used to determine the refractive power of the intraocular lenses retrospectively and to calculate the possible refractive outcome. RESULTS: Precision of partial coherence interferometry biometry was more than 10 times better than that of ultrasound. Therefore, the possible mean absolute error for postoperative refraction achieved with partial coherence interferometry biometry was 0.49 diopters (compared with 0.67 diopters with ultrasound biometry), resulting in an improvement of 27%. Axial eye length measured with the two techniques differed by a mean of 460 microm. The difference in lens thickness measured with partial coherence interferometry and ultrasound significantly correlated with cataract grade. A mean shortening of 120 microm of axial eye length following cataract surgery was also detected by partial coherence interferometry. CONCLUSIONS: The enhanced version of partial coherence interferometry offers biometry with unprecedented precision (<10 microm) and resolution (approximately 12 microm), therefore improving the refractive outcome in cataract surgery. This noninvasive technique provides a high degree of comfort for the patient, with no need for local anesthesia or pupil dilation and minimized risk of corneal infection.

Eye elongation during accommodation in humans: differences between emmetropes and myopes.

PURPOSE: The pathophysiology and pathogenesis of myopia are still a matter of controversy. Exaggerated longitudinal eye growth is assumed to play an important role in the development of myopia. A significant correlation between refraction and amount of near-work has been reported. However, current knowledge of changes of axial eye length with accommodation is limited because clinical ultrasound biometry does not provide the precision and resolution required to thoroughly investigate these phenomena. METHODS: Partial coherence interferometry (PCI), a noninvasive biometric technique, uses laser light with short coherence length in combination with interferometry to achieve precision in the micrometer to submicrometer range and resolution of 10 microm. In the present study this technique was used to investigate axial eye length changes in 11 emmetropic and 12 myopic eyes during monocular fixation at the far and near point. In 7 subjects, the contralateral eye has also been measured to investigate interocular differences in eye elongation. RESULTS: All investigated eyes elongated during accommodation. This elongation was more pronounced in emmetropes than in myopes (P < 0.001). Mean accommodation-induced eye elongations of 12.7 microm (range, 8.6-19.2 microm) and 5.2 microm (range, 2.1-9.5 microm), corresponding to a dioptric change of approximately -0.036 D and -0.015 D, were obtained for emmetropes and myopes. No significant difference in accommodative amplitudes between groups (5.1 +/- 1.2 D [range, 3.8-7.1 D] versus 4.1 +/- 2.0 D [range, 1.0-7.1 D]; P = 0.14) was detected. No significant interocular difference in accommodation-induced eye elongation was revealed (P = 0.86). Also, a mean backward movement of the posterior lens pole of 38 microm (range, 9-107 microm) was observed in both study groups. CONCLUSIONS: The detected eye elongation can be explained by the accommodation-induced contraction of the ciliary muscle, which results in forward and inward pulling of the choroid, thus decreasing the circumference of the sclera, and leads to an elongation of the axial eye length. Finally, it was demonstrated that PCI, in contrast to clinical ultrasound, is capable of characterizing eye length changes during accommodation in humans.

High precision biometry of pseudophakic eyes using partial coherence interferometry.

PURPOSE: To investigate the applicability of the scanning version of dual-beam partial coherence interferometry (PCI) for measuring the anterior segment and axial length of pseudophakic eyes in a clinical setting and to determine the achievable precision with this biometry technique. SETTING: Department of Ophthalmology, Vienna General Hospital, and Institute of Medical Physics, University of Vienna, Austria. METHODS: Partial coherence interferometry was performed in 39 pseudophakic eyes of 39 patients after implantation of a foldable acrylic intraocular lens (IOL). RESULTS: Effective lens position (ELP), IOL thickness and lens-capsule distance (LCD) were determined with a precision of 2 to 3 microns; corneal thickness and axial eye length, with a precision of 0.8 and 5.0 microns, respectively. The mean ELP of the IOL was 4.093 mm +/- 0.290 (SD). In 7 eyes (18%), a positive LCD of 68 +/- 40 microns was detected with PCI. Mean corneal thickness was 526.4 +/- 31.5 microns; mean IOL thickness, 791.5 +/- 40.2 microns; and mean axial length, 23.388 +/- 0.824 mm. CONCLUSION: The scanning version of PCI enables high precision (< or = 5 microns) and high resolution (approximately 12 microns) biometry of pseudophakic eyes that is better than conventional ultrasound by a factor of more than 20. For the first time, positive LCD, a possible risk factor for posterior capsule opacification, could be detected and quantified. Furthermore, this technique offers a high degree of comfort for the patient since it is a noncontact method with no need for local anesthesia or pupil dilation and has a reduced risk of corneal infection.

Accurate determination of effective lens position and lens-capsule distance with 4 intraocular lenses.

PURPOSE: To measure effective lens position (ELP) of 4 intraocular lenses (IOLs) using high precision and high resolution dual-beam partial coherence interferometry (PCI) and to assess the tendency of these IOLs to produce a lens-capsule distance (LCD), a possible risk factor for posterior capsule opacification. SETTING: Department of Ophthalmology, Vienna General Hospital; Institute of Medical Physics, University of Vienna, Austria. METHODS: In a retrospective study, PCI was used to measure ELP and LCD in 139 pseudophakic eyes of 110 patients with 4 IOLs: acrylic 3-piece IOL (AcrySof MA60BM); silicone 3-piece IOL without a capsular tension ring (PhacoFlex SI30) and with a capsular tension ring (PhacoFlex SI30 and Morcher Type 14); silicone plate-haptic IOL (Staar AA4203VF); and a hydrogel plate-haptic IOL (logel 1103). RESULTS: The ELP and LCD were determined with a precision of approximately 3 to 4 microns. An LCD was detected in 21% eyes with the AcrySof, 20% of eyes with the SI30 without a capsular tension ring, 10% of eyes with a capsular tension ring, 21% of eyes with the Staar, and 17% of eyes with the logel. The LCDs detected by PCI, but not by slitlamp examination, were significantly smaller than those detected by both. CONCLUSION: The amount of LCD detected by PCI was approximately the same with all IOL types (approximately 20%) except the PhacoFlex SI30 with a capsular tension ring (10%).

Investigation of dispersion effects in ocular media by multiple wavelength partial coherence interferometry.

We report on quantitative measurements of group refractive indices and group dispersion in water and in human ocular media such as the cornea, the aqueous humor, the lens, artificial intraocular lenses, as well as a total value averaged over the media along the axial eye length of normal subjects and pseudophakic patients in vivo using dual beam partial coherence interferometry. Different optical thickness values due to the dispersion of the cornea are demonstrated using two spectrally displaced light sources. The displacement can be used to indirectly calculate the group dispersion of the human cornea in the spectral region between 810 nm and 860 nm. If the object under investigation is dispersive, resolution is limited due to a broadening of the detected signals. This broadening increases with group dispersion, i.e., the extent to which the group refractive index of the medium varies with wavelength and thickness of the tissue under investigation as well as with the spectral bandwidth of the light source. Measurements of the group dispersion in the cornea, lens and vitreous of pseudophakic and normal human eyes, show that the cornea and the lens are more dispersive than water-by a factor of about 5 and 2, respectively-in the investigated spectral region. The cornea is approximately threefold more dispersive than the human crystalline lens, the aqueous humor is less dispersive than water and the group dispersion of all ocular components together, averaged over the axial length of normal and pseudophakic eyes, was only slightly higher compared to that of water. Since the highly dispersive cornea and lens together have only a thickness of about one sixth of that of the axial eye length, it seems that their contribution to the group dispersive effect along the whole axial eye length is only small.

Signal and resolution enhancements in dual beam optical coherence tomography of the human eye.

In the past 10 years, a dual beam version of partial coherence interferometry has been developed for measuring intraocular distances in vivo with a precision on the order of 0.3 to 3 µm. Two improvements of this technology are described. A special diffractive optical element allows matching of the wavefronts of the divergent beam reflected at the cornea and the parallel beam reflected at the retina and collimated by the optic system of the eye. In this way, the power of the light oscillations of the interfering beams incident on the photodetector is increased and the signal-to-noise ratio of in vivo measurements to the human retina is improved by 20 to 25 dB. By using a synthesized light source consisting of two spectrally displaced superluminescent diodes with an effective bandwidth of 50 nm, and by compensating for the dispersive effects of the ocular media, it was possible to record the first optical coherence tomogram of the retina of a human eye in vivo with an axial resolution of ∼6 to 7 µm. This is a twofold improvement over the current technology. © 1998 Society of Photo-Optical Instrumentation Engineers.