Optical resolution, the ability to detect the presence of two closely spaced points as being physically different from a single point, has many potential definitions. Light emanating from a point in the object space is imaged, not as a point, but as a distribution of light over an area. The PSF describes the characteristic of this spot. One standard for measuring resolution is to consider the image of a point source to be an Airy disk and the limits of resolution to be where the maximum of the Airy disk from one source lies in the first minimum of the diffraction pattern in the Airy disk of another source, and this is known as the Rayleigh criterion. In systems using light originating from a point source, such as OCT or confocal microscopy, the Airy disk may be difficult to measure. Instead, the FWHM of the light distribution is used as it is relatively easy to measure and represents the lateral PSF of the illumination system. In confocal imaging, there is a radial reduction in the intensity of the illumination and a corresponding radial reduction in the sensitivity of detection. This apodization removes the influences of potential side-lobes in the illumination and detection systems’ optical performance and shapes the lateral point spread function of confocal imaging systems. Thus, the PSF of the system, rather than just the spot size on the retina or just the lateral PSF of the detection system, becomes the paramount metric to evaluate lateral resolution produced in the image. The estimates of the lateral PSF of the OCT instrument, the Heidelberg Spectralis, from particles resulted in an estimate of a FWHM of 5.11 µm for diamond powder and 4.9 µm for the FeO target. The Fourier analysis, which estimates the diffraction limited performance, yielded 5.0 µm and the OpticStudio modeling produced the estimate of 4.6 um. These results are comparable, and each method has its own limitations, as will be discussed below. The USAF target, which has been used to estimate lateral resolution of OCT instruments, showed a lateral resolution of 4.6 µm.
The diamond powder target was prepared by mixing the 0 to 1 µm diamond particles with silicone elastomer before it polymerized. With extensive stirring, the diamond particles are separated to produce individually reflecting grains. It is possible that small aggregates of grains were present in the test sample and imaged as one particle. To avoid inclusion of large aggregates, we limited the analysis to particle signals whose FWHM lies within three standard deviations of the overall mean. The mean PSF subsequently calculated from diamond powder was 4.7% larger than that measured from a validated sample containing FeO particles that were 800 nm in diameter. Possible reasons for the difference include a greater tendency toward clumping with the diamond particles, a larger distribution in the size of the diamond grains, and possible inaccuracies in the size estimation of the diamond grains. The distribution of the intensity of the reflections showed tails of the distribution that were slightly more elevated from zero with the diamond particles than for the FeO particles, suggesting a wider range of particle sizes in the diamond powder sample (see
Fig. 3). We did not independently measure the size of the diamond powder grains or the resultant suspended particles. In the sequence in the measurements, the diamond particles were measured first and then a validated test object using FeO was evaluated, which confirmed and potentially refined the measurements made from the diamond particles.
Images derived from human eyes with no pharmacologic dilation were used to determine the magnitudes of the complex ATF. Using this analysis, we calculated the diffraction limited resolution to be approximately 5.0 µm. This is a theoretical value that has an uncertain relationship with actual resolution in the eye, which can have significant aberrations. The relative proportions of diffraction versus aberrations in limiting optical performance are related to pupil sizes. In eyes with smaller pupils, diffraction limits the optical performance, whereas at large pupil sizes, aberrations dominate. Given the 1/e2 size of 1.71 mm, diffraction may play a role in the lateral resolution of OCT in clinical practice, particularly in patients with undilated pupils.
The 1951 USAF target was used for estimation of the lateral resolution. This target has two groups of triplets of bars for each resolution arranged orthogonal to each other. We measured the resolution along the fast axis of the scan. The resultant reflectance profiles showed decreasing contrast with increasing spatial frequency, as would be expected. There are differing methods of inferring image resolution; with the Rayleigh criterion the contrast ratio is 26.4%. There is the Sparrow criterion, which is essentially 0% and other criteria for resolution lying in between. Thus, we show the elements in the USAF chart and their corresponding contrast measurements. Because there were easily discernable variations in the brightness in group 6, element 4, but not in element 5, the resolution was calculated to be 4.6 µm in the eye. The USAF target was established for incoherent imaging, and there is a possibility that in a coherent regime there could be destructive or constructive interference produced by reflections from between the dark and light regions. This has the possibility of creating bars with lighter or darker appearance than what they would have been using an incoherent imaging method, and thereby bias the true estimate of image resolution. If the flat surface is scanned with a focused beam, then the local angle of incidence inevitably varies across the target. The mirrored beam is not reflected exactly into itself across much of the target, so that depending on the scan position, only a certain portion of the numerical aperture (NA) is actually used. We include the USAF evaluation here because of its historical use.
13–20
The various test modalities used in this study all consistently point to a PSF of the instrument being much better than the beam width on the retina, quoted to be 14 µm for the Spectralis. As currently implemented in the commercial instrument, the actual resolution of the instrument was not realized because the tissue sampling was done at an interval to achieve 14 µm lateral resolution, that is, a 6 µm A-scan spacing. The lower resolution expectation was cemented in place by engineering designed to achieve only the lower lateral resolution. To recover the inherent resolution capabilities of the instrument a scan spacing of 1.9 µm was used in the present study. This sampling is slightly more than three times the “High Resolution” mode of the commercially available instrument. The original A-scan rate of the instrument was selected as an optimization among costs at the time, the lateral resolution as it was understood, and convenience for the operator and patient in terms of scan times. Just increasing the scan density by a factor of three through a software modification would not incur significant hardware costs and would help achieve better lateral resolution. In practical use, increasing the scan density would create a much larger burden on the patient, operator, and eye tracking system because of the necessary increase in scanning time and precision required in maintaining image alignment. To realize the true potential lateral resolution with a convenient scanning time, a much higher A-scan rate would be necessary. Tighter tolerances in the slow axis scanning to improve the positioning of the adjacent B-scans would be required as well for an optimal system. Although this study of the disparity between stated spot size on the retina and the actual achievable PSF was measured in the Heidelberg Spectralis, there are reasons to believe these same factors likely apply to every OCT device in the marketplace. It may be possible that all available instruments under sample the tissue. Increasing the A-scan rates of commercial OCT instruments has been a trend for decades, and part of the motivation is to be able to scan increasingly large areas. The increased A-scan rates may also be used to configure scanning protocols that avoid under sampling to improve achievable lateral resolution available in the clinic.
How we image these structures is a function of past translational efforts of converting theoretical theories into clinical practice. The resultant images obtained helped form our current concepts of the retina and choroid in health and disease. The determinants of lateral resolution proposed many years ago, which seemed to have forged currently used scanning parameters, do not appear to be correct. The actual lateral resolution of OCT appears to be much better than previously proposed, with no structural change in the scanner necessary. Part of translational research is the testing longstanding assumptions and by improving resolution may help drive development of improved understanding of retinal physiology and disease in the future. These changes would appear to apply to current commercial OCT instruments by all manufacturers, as their stated lateral resolution is generally between 15 and 25 µm. By the same logic used in our analysis, it is likely the actual lateral resolution of these instruments is potentially much better (
Table 2).