In the early stages of some retinal diseases, such as glaucoma, loss of retinal activity may be difficult to detect with today's clinical instruments. Many of today's instruments focus on detecting changes in anatomical structures, such as the nerve fiber layer. Our device, which is based on a modified fundus camera, seeks to detect changes in optical signals that reflect functional changes in the retina. The functional imager uses a patterned stimulus at wavelength of 535nm. An intrinsic functional signal is collected at a near infrared wavelength. Measured changes in reflectance in response to the visual stimulus are on the order of 0.1% to 1% of the total reflected intensity level, which makes the functional signal difficult to detect by standard methods because it is masked by other physiological signals and by imaging system noise. In this paper, we analyze the video sequences from a set of 60 experiments with different patterned stimuli from cats. Using a set of statistical techniques known as Independent Component Analysis (ICA), we estimate the signals present in the videos. Through controlled simulation experiments, we quantify the limits of signal strength in order to detect the physiological signal of interest. The results of the analysis show that, in principle, signal levels of 0.1% (-30dB) can be detected. The study found that in 86% of the animal experiments the patterned stimuli effects on the retina can be detected and extracted. The analysis of the different responses extracted from the videos can give an insight of the functional processes present during the stimulation of the retina.
An optical imaging device of retina function (OID-RF) has been developed to measure changes in blood oxygen saturation due to neural activity resulting from visual stimulation of the photoreceptors in the human retina. The video data that are collected represent a mixture of the functional signal in response to the retinal activation and other signals from undetermined physiological activity. Measured changes in reflectance in response to the visual stimulus are on the order of 0.1% to 1.0% of the total reflected intensity level which makes the functional signal difficult to detect by standard methods since it is masked by the other signals that are present. In this paper, we apply principal component analysis (PCA), blind source separation (BSS), using Extended Spatial Decorrelation (ESD) and independent component analysis (ICA) using the Fast-ICA algorithm to extract the functional signal from the retinal videos. The results revealed that the functional signal in a stimulated retina can be detected through the application of some of these techniques.
To overcome the difficulty in detection of loss of retinal activity, a functional-Retinal Imaging Device (f-RID) was developed. The device, which is based on a modified fundus camera, seeks to detect changes in optical signals that reflect functional changes in the retina. Measured changes in reflectance in response to the visual stimulus are on the order of 0.1% to 1% of the total reflected intensity level, which makes the functional signal difficult to detect by standard methods because it is masked by other physiological signals and by noise. In this paper, we present a new Independent Component Analysis (ICA) algorithm used to analyze the video sequences from a set of experiments with different patterned stimuli from cats and humans. The ICA algorithm with priors (ICA-P) uses information about the stimulation paradigms to increase the signal detection thresholds when compared to traditional ICA algorithms. The results of the analysis show that we can detect signal levels as low as 0.01% of the total reflected intensity. Also, improvement of up to 30dB in signal detection over traditional ICA algorithms is achieved. The study found that in more than 80% of the in-vivo experiments the patterned stimuli effects on the retina can be detected and extracted.
An optical imaging device of retina function (OID-RF) has been constructed to record changes in reflected 700-nm light from the fundus caused by retinal activation in response to a visual 535-nm stimulus. The resulting images reveal areas of the retina activated by visual stimulation. This device is a modified fundus camera designed to provide a patterned, moving visual stimulus over a 45-degree field of view to the subject in the green wavelength portion of the visual spectrum while simultaneously imaging the fundus in another, longer wavelength range. Data was collected from 3 normal subjects and recorded for 13 seconds at 4 Hz; 3 seconds were recorded during pre-stimulus baseline, 5 seconds during the stimulus, and 5 seconds post-stimulus. This procedure was repeated several times and, after image registration, the images were averaged to improve signal to noise. The change in reflected intensity from the retina due to the stimulus was then calculated by comparison to the pre-stimulus state. Reflected intensity from areas of stimulated retina began to increase steadily within 1 second after stimulus onset and decayed after stimulus offset. These results indicated that a functional optical signal can be recorded from the human eye.
Imaging studies from anesthetized feline, primate, and human retinas have revealed near-infrared fundus reflectance changes induced by visible light stimulation. In the present study, the spatial and temporal properties of similar changes were characterized in normal, awake humans.Five normal human subjects were studied. A modified fundus camera was used to image changes in retinal reflectance of 780-nm near-infrared light imaged onto a 12-bit charge-coupled device (CCD) camera in response to a green (540 nm) visual stimulus. During 60 seconds of recording (frame rate, 3 Hz) 10 cycles were recorded, during each of which 3 seconds of blank and then 3 seconds of either vertical bar or blank stimulus was projected. The change in the average near-infrared reflectance of the stimulated retinal region relative to an equal-sized nonstimulated region (r is the ratio of reflectance between the two retinal areas) was analyzed with a mixed model for repeated measures.The mixed model showed a significant average decrease in r of 0.14% (95% CI, -0.25 to -0.03) over all subjects induced by bar stimulus cycles, with a gradual return to baseline after stimulus offset, compared with only a 0.04% (95% CI, -0.11-+0.20) decrease in r induced by blank, nonstimulated cycles. The mixed model for individuals showed a decreasing linear trend in r over time during bar stimulation, but no decrease for blank cycles in three of five subjects.There was a localized decrease in reflectance in response to 780-nm near-infrared light in the retinal region exposed to a visual stimulus, which was significant in three of five subjects. It is presumed that the reflectance change represents the functional activity of the retina in response to a visual stimulus.
Non-invasive imaging of retinal function based on the recording of spatially distributed reflectance changes evoked by visual stimuli has to-date been performed primarily using modified commercial fundus cameras. We have constructed a prototype retinal functional imager, using a commercial endoscope (Storz) for the frontend optics, and a low-cost back-end that includes the needed dichroic beam splitter to separate the stimulus path from the imaging path. This device has been tested to demonstrate its performance for the delivery of adequate near infrared (NIR) illumination, intensity of the visual stimulus and reflectance return in the imaging path. The current device was found to be capable of imaging reflectance changes of 0.1%, similar to that observable using the modified commercial fundus camera approach. The visual stimulus (a 505nm spot of 0.5secs) was used with an interrogation illumination of 780nm, and a sequence of imaged captured. At each pixel, the imaged signal was subtracted and normalized by the baseline reflectance, so that the measurement was ΔR/R. The typical retinal activity signal observed had a ΔR/R of 0.3-1.0%. The noise levels were measured when no stimulus was applied and found to vary between ± 0.05%. Functional imaging has been suggested as a means to provide objective information on retina function that may be a preclinical indicator of ocular diseases, such as age-related macular degeneration (AMD), glaucoma, and diabetic retinopathy. The endoscopic approach promises to yield a significantly more economical retinal functional imaging device that would be clinically important.
