Microelectrode depth study of the electroretinographic oscillatory potentials in the frog retina

The depth profile of the electroretinographic oscillatory potentials was studied in the isolated frog retina. The intraretinal electrode was introduced from the receptor side, and the reference electrode was placed on the vitreal side. The electroretinogram, recorded either transretinally or with the electrode tip in the receptor layer, showed 4 to 10 oscillatory potential wavelets. As the electrode was advanced proximally, the wavelets disappeared as a function of retinal depth. The wavelets with longer peak latencies disappeared earlier, and only the first one or two wavelets could be identified when the electrode was in the inner plexiform layer. These findings indicate that the oscillatory potentials are generated between the inner and outer plexiform layers and that the earlier wavelets originate in the more proximal retina. The results are consistent with the notion that the oscillatory potentials are generated by synaptic feedback circuits.     

Performance of the DTL electrode compared to the Jet contact lens electrode in clinical testing

To compare the recording characteristics of the DTL fiber and Jet contact lens electrodes, ERG responses were recorded from 20 normal subjects using the ISCEV standard clinical protocol. In each subject, the DTL electrode was placed in the right eye and the Jet contact lens in the left eye after 30 min of dark-adaptation. After presenting standard dark- and light-adapted stimuli, each response was analyzed for a- and b-wave amplitude and implicit time. Each subject was then asked to rate the comfort of each electrode. Responses recorded with the DTL are larger and faster for the dark-adapted dim white and red flashes. In contrast, the Jet electrode records larger responses for the dark-adapted standard flash and light-adapted flashes. Of these differences, only the standard flash is statistically significant. However, overall the DTL electrode has greater variability than the Jet electrode across all recordings. The DTL and Jet electrodes cannot be used interchangeably as demonstrated by a Bland and Altman analysis of agreement. Thus, separate normative values need to be collected for each type of electrode to accurately assess retinal function. Based on the comfort assessment of the two electrodes, the subjects preferred the DTL electrode over the Jet electrode. Due to the superior comfort level of the DTL and its similarities to the Jet contact lens electrode in ERG recordings, the DTL electrode appears to be a good alternative to the Jet electrode in routine clinical testing when used with the appropriate normative values.     

The effect of iodoacetic acid on the electroretinogram and oscillatory potentials in rabbits

It has been shown that a single injection of iodoacetic acid selectively (but temporarily) abolishes the b-wave of the electroretinogram. We examined whether such use of this chemical further substantiate our claim that the b-wave of the electroretinogram is a composite potential resulting from the summation (or integration) of faster retinal potentials, usually referred to as the oscillatory potentials. Full-field electroretinograms were recorded from adult New Zealand rabbits before and after a single, bolus injection of 15mg/kg of buffered iodoacetic acid. Both the 1–1000 Hz electroretinogram and the 100–1000 Hz oscillatory potentials were recorded simultaneously. The oscillatory potentials considered in this study were those normally seen on the rising phase of the b-wave. Following the intravenous injection of iodoacetic acid, there was a progressive decrease in the amplitude and peak time of the b-wave. This observation also was reflected in the oscillatory potential recordings, in which the long-latency oscillatory potentials (3 and 4) progressively disappeared while oscillatory potential 2 remained. We believe that these findings further support our contention that the oscillatory potentials are major components of the b-wave.     

The repeatability and variability of the multifocal electroretinogram for four different electrodes

Multifocal electroretinograms were recorded from twelve subjects on three separate days using the Visual Evoked Response Imaging System (VERIS). The subjects were 18–40 years-of-age. Each recording session was separated by an interval of between one and 16 weeks. Four electrode types were compared: the JET contact lens electrode, the gold foil electrode, the DTL thread electrode and the carbon fibre (c-glide) electrode. The b-wave amplitude density (nV/ sq.deg) recorded with each electrode was measured. The mean amplitude density recorded at each session for each subject and each electrode was calculated and compared. There were no significant differences in amplitude density for the ERGs recorded with each type of electrode over the three days. The coefficient of variation for each electrode and each subject was then calculated using the results for the three days. The coefficients of variation for each electrode showed significant differences (One-way ANOVA: F =4.51. d.f. =3, 44. P < 0.008). The c-glide electrode results had the highest variability and post hoc tests showed that its coefficient of variation was significantly different from those of the JET and gold foil electrodes but not from that of the DTL thread electrode.     

Inter-subject,inter-ocular and inter-session repeatability of the photopic negative response of the electroretinogram recorded using DTL and skin electrodes

The photopic negative response (PhNR) has attracted interest as a flash ERG component reflecting inner retinal activity, with investigators adopting various approaches to analysing the response. This study has two principal aims: first to determine the most reliable technique for assessing the PhNR amplitude; secondly to compare the repeatability characteristics of the PhNR recorded using DTL and skin active electrodes. Electroretinograms were recorded in 31 subjects, using both electrode types, in response to a Ganzfeld red stimulus (Lee filter “bright red”; 1.76 log phot td.s; 4 Hz) presented over a steady blue background (Schott glass filter BG28; 3.9 log scot td). Sixteen subjects returned to assess repeatability. PhNR amplitude was measured from b-wave peak-to-PhNR trough, pre-stimulus baseline to trough, and from peak and baseline to a fixed time-point; a ratio of b-wave/PhNR amplitude was also calculated. Coefficients of variation (CoV), and inter-ocular and inter-session limits of agreement (LoA) were calculated for all measures. The ratio of b-wave/PhNR amplitude showed the lowest CoV (14.3% DTL; 23.2% skin), inter-ocular LoA (22.2% DTL; 25.0% skin), and inter-session LoA (22.8% DTL; 20.3% skin). The peak-to-trough and peak-to-fixed-time measurements were also consistently reliable. Least reliable measures were those measured from baseline. While skin electrode responses were significantly smaller than DTL responses (P < 0.0001), the variability was only slightly increased. This study suggests that peak-to-trough measurements are the most reliable means of measuring the PhNR and ratio calculation further improves repeatability. Skin electrodes provided a viable alternative to DTL electrodes for recording the PhNR.     

Evaluation of different recording parameters to establish a standard for flash electroretinography in rodents.

Different electrodes and stimulus protocols commonly used for electroretinography in rodent eyes were compared for convenience of use, degree of damage to corneal epithelium, and for magnitude of amplitude, reproducibility, left versus right eye accuracy, and reliability of recorded parameters of the flash electroretinogram (ERG). Adult C57BL/6 pigmented mice and albino Wistar rats were used to determine scotopic ERGs in response to Ganzfeld or strobe-light stimulation and light-adapted (photopic) ERGs recorded from both eyes at the same time. Test-retest data were used for statistical analyses to compare a monopolar gold-wire contact lens electrode (CLE), a cotton-wick silver-silver chloride electrode (CSCE), a DTL fiber electrode (DTLE), and a circular stainless steel wire electrode (SSE). Corneas were evaluated for abrasion after ERG recordings using fluorescein staining and also for the time taken, ease of insertion, and re-insertions required for the different electrodes. Compared to CSCE, DTLE, and SSE, the ERG potentials recorded by CLE had significantly larger scotopic amplitudes and oscillatory potentials under strobe or Ganzfeld stimulation and for light-adapted ERG b-wave amplitudes in both mice and rats. In analyzing test-retest data of scotopic ERG a-wave and b-wave amplitudes, the intraclass correlation coefficient showed the best agreement for the CLE (range 0.61-0.94) compared to the SSE (0.13-0.77), DTLE (0.02-0.69), and CSCE (0.12-0.51). In mice and rats, logistic regression analyses revealed significant correlations for amplitudes of most scotopic ERG parameters between contralateral eyes obtained with CLE and for some ERG components recorded by SSE. When comparing ERG amplitudes for stimulation by strobe or Ganzfeld, the difference was least with the CLE compared to DTLE, CSCE, or SSE. The time taken to insert the four different electrodes was greatest for the CLE in both mice and rats. The extent of corneal abrasion resulting from electrode use in mice was largest for the SSE followed by the CLE. However, in rats there was almost no corneal damage after ERG recordings with the CLE. Because of the stability of eye contact, the CLE allows ERGs to be determined over a longer recording session. Recording of scotopic and photopic (light-adapted) ERGs in rodents with monopolar gold-wire contact lens electrodes provides greater amplitudes and higher reproducibility when compared to other commonly used corneal electrodes. These electrodes are significantly better overall than others that were evaluated and should be considered for a standard protocol to monitor retinal function in rodent eyes.     

不同多焦视网膜电图系统在评估视网膜功能上的比较

目的 评估VERIS mfERG （EDI Inc）和Espion multifocal imager （Diagnosys）两种不同系统测量正常眼多焦视网膜电图（mfERG）的电位波反应的差异及其相关性,并用两个不同系统分别测量黄斑前膜患眼的mfERG并进行分析.方法 这是一项横断面研究,在VERIS系统使用Burian-Allen （BA）电极,Espion系统使用BA和DTL电极分别测量25只正常眼的mfERG,将3次测得的一阶反应的相应数据进行配对t检验及Pearson相关分析.两个系统用BA电极分别测量19只黄斑前膜患眼的mfERG,对一阶反应的相应数据进行Fisher＇s exact test分析.结果 VERIS BA测得正常眼N1波、P1波振幅密度值最大,其次为Espion BA,Espion DTL最小;VERIS BA测得N1波、P1波潜伏期最长,Espion BA与Espion DTL差异无统计学意义.Espion BA与VERIS BA测得的P1波振幅密度、潜伏期皆有明显的相关性.黄斑前膜患眼中,VERIS BA比Espion BA能检测到更多P1波振幅密度和P1波潜伏期的异常.结论 不论VERIS或Espion系统都能成功记录多焦视网膜电图,当两种系统都使用Burian-Allen电极时,两者mfERG一阶反应P1波振幅密度及潜伏期有着良好的相关性.     

Comparison of performance and patient satisfaction of two types of ERG electrodes

BACKGROUND: An age-controlled comparison concerning patient satisfaction and electrical performance of microfibres (DTL) and rigid contact lens (Henkes) corneal ERG electrodes was carried out. METHODS: 36 test persons underwent complete ophthalmological examination and were equally distributed into 3 age groups. Electroretinograms were recorded according to ISCEV standards. Randomly, in one eye a Henkes electrode was used and in the other eye a DTL electrode. Amplitudes of a- and b-waves and implicit times were measured and compared for the two electrode types. RESULTS: 34 of 36 test persons preferred DTL electrodes. Electrical performance concerning b-wave amplitudes was comparable. Statistically significant differences were detected only for scotopic combined cone-rod stimulation in the age groups 20 - 40 and 41 - 60 years between the different electrodes. Other recordings did not show differences. A statistically significant reduction of signal amplitudes with age was detected for scotopic isolated rod signals and combined cone-rod signals. Significance level was p < 0.05. No conjunctival or corneal erosions were found after ERG recordings for either electrode. CONCLUSIONS: Electrical performance is comparable between electrodes. For scotopic stimulations age was a significant influencing factor for signal amplitude and should be respected for normative values. DTL electrodes were preferred by the vast majority of patients. No adverse clinical effects were observed for either electrode. DTL electrodes should be preferred due to hygienic reasons (single use) and patient comfort.     

Maturation of the electroretinogram of the neonatal rabbit

The development of the electroretinogram in the rabbit has been said to proceed with first the a-wave, then the b-wave and last the oscillatory potentials. The aim of our study was to reexamine this claim with special attention to the oscillatory potentials. Albino rabbits from the same litter were studied at weekly intervals for five weeks from the first week of life. A Grass photostimulator was used in light and dark adaptation and 50 amplified responses were averaged. Both 1–1000 Hz (electroretinogram) and 100–1000 Hz (oscillatory potential) band-widths were recorded simultaneously. The a-wave was the earliest signal to appear, at the second week of life. A rapid growth of the b-wave and oscillatory potentials was then noted between the second and third weeks, followed by a slower change. They evolved at the same rate, each with an increase in amplitude and decrease in peak time. The change in form of the b-wave was consistent with the sequential formation of each potential as the rabbit matured. The finding of simultaneous development of the b-wave and oscillatory potentials in the aging neonatal rabbit is contrary to previous reports.     

Abnormalities of the electroretinogram and visual-evoked potential in vitamin E deficient rats.

Flash electroretinograms, retinal oscillatory and cortical visual-evoked potentials were recorded in 10-month vitamin E deficient rats and in age-matched controls. A significant increase in the latency (P less than 0.0001) and decrease in amplitude (P less than 0.0001) of the electroretinogram a- and b-waves were observed in the vitamin E deficient rats compared with controls. The vitamin E deficient rats also showed a significant delay (P less than 0.002) in early oscillatory potentials. No significant group differences were obtained in the P1 latency or P1-N1 amplitude of the visual-evoked potential; however, the N1 peak was significantly delayed (P = 0.01) in the vitamin E deficient rats. This study shows, for the first time, that the electroretinogram and visual-evoked potential to flash stimulation provide a sensitive index for monitoring the visual effects of vitamin E deficiency in the rat.     

Safety and efficacy evaluation of a new ERG electrode (the LVP electrode); Part II. Flash ERG pilot study

A new disposable, non-contact electrode (LVP electrode) was evaluated for use in recording flash ERG. A comparison of DTL and LVP electrodes showed reproducible waveforms for all components analysed. There was a good correlation obtained between the DTL and LVP electrodes. The LVP electrode is safe, less expensive and has less movement of the fiber due to its rigidity.     

The pattern reversal VEP in short-gestation infants on taurine or taurine-free diet

The visual evoked potential to flash stimulation has been studied in premature infants for many years. There have been no studies of the response to pattern reversal stimuli during the pre-term period, however. Twenty babies all born prematurely between 32 and 35 weeks post menstrual age had pattern reversal visual evoked potentials recorded on at least two occasions between the ages of 32.5 and 52 weeks post menstrual age with the first recording prior to reaching 40 weeks post menstrual age. The stimulator consisted of a small hand-held television with a 55 × 40 mm screen, and responses were averaged by a Cadwell 5200 averager. The check size was 7 × 7 mm and the contrast was 78%. The stimulator was held at a distance of 20cm from the eyes. The angular subtense of the TV screen was 15.3 ° × 11.31 °. On each occasion flash visual evoked potentials were obtained for comparison, as were flash electroretinograms recorded with a DTL electrode. The infants were randomly assigned to receive breast milk substitute with or without taurine.Pattern reversal responses consisted of a positive component with a mean latency of 320 msec at 32.5 weeks post menstrual age. The negative relationship between age at recording and the latency of the major positive component of the pattern reversal visual evoked potential was significant at p < 0.0001. There was no apparent difference in the visual responses of the two groups.Abbreviations OP Oscillatory Potential - PMA Postmenstrual Age     

Influence of Recording Electrode Type and Reference Electrode Position on the Canine Electroretinogram

Electroretinography is commonly used to assess the functional integrity of the retina. There are many external variables that can influence the electroretinographic waveforms recorded, and it is important to be aware of these so as not to misinterpret their effects as abnormalities in retinal function. In this study we examined the effect of three different recording electrodes on the ERGs recorded from normal dogs. A bipolar Burian–Allen lens, a monopolar Dawson Trick Litzkow (DTL) fiber electrode, and a monopolar ERG-Jet lens electrode were compared. The effect of altering the distance of the reference electrode from the eye was also examined; using the ERG-Jet lens electrode, the ERG was recorded with the reference electrode placed over the zygomatic arch at 1, 3 and 5 cm caudal to the lateral canthus. The ERGs recorded with the bipolar Burian–Allen lens had significantly lower amplitudes, higher a-wave thresholds and a shallower initial a-wave slope, than those recorded by the two monopolar electrodes. Positioning the reference electrode further from the eye resulted in significantly higher amplitudes. Naka-Rushton fitting and calculation of retinal sensitivity (K) gave significantly different results between the Burian–Allen lens and ERG-Jet lens electrode with the reference electrode 5 cm from the lateral canthus. These results demonstrate that recording electrode type and distance of the reference electrode from the eye significantly affect the ERG tracings of the dog, and may alter the assessment of retinal function that can therefore be derived. Results obtained using these three different types of electrodes cannot be directly compared.     

The importance of electrode position in visual electrophysiology

Purpose

The DTL fibre electrode is commonly used to record the electric potentials elicited by stimulation of the retina. Two positions are commonly used: it is placed either on the cornea along the lower lid or in the conjunctival fornix. The PERG and OPs have previously been examined and compared under both conditions. The aim of this study was to examine the ERG, flicker response and on–off responses with differing electrode positions.

Methods

Before recruitment, all subjects underwent an ophthalmological examination. We enrolled 13 normal control subjects into the study aged 13–64 years, all with a visual acuity of ≥1.0. We recorded scotopic and photopic ERGs, flicker and on–off responses, for both electrode positions. On the first day, one eye had the electrode placed on the cornea along the lower lid and the other eye had it positioned in the conjunctival sac. On a second day, the recordings were repeated with the alternative electrode placements.

Results

ERG, on–off and flicker responses were all smaller by between 20 and 25% when the DTL electrode was positioned in the conjunctival sac, compared to when it was positioned on the cornea, as did the scatter in the data points. This indicates that there is no advantage clinically for one or the other placement.

Conclusions

Our results confirm other reports examining the effect of electrode position on electrophysiological potentials. When recording with the DTL electrode, it is important to ensure that it is placed at the same position in repeat recordings or in multicentre trials and that it is stable and does not move during recording.

     

Reproducibility of ERG responses obtained with the DTL electrode.

Previous investigators have suggested that the DTL fibre electrode might not be suitable for the recording of replicable electroretinograms. We present experimental evidence that when used adequately, this electrode does permit the recording of highly reproducible retinal potentials.     

Electroretinographic evaluation in adult diabetics

In cross-sectional fashion, we recorded the maximal combined response and 30-Hz flicker responses in 178 adult diabetics and 40 normal controls according to the recommendations of the International Society of Clinical Electrophysiology of Vision. The oscillatory potentials were extracted from the maximal combined response by high-pass filtering. The clear media and attached retina were criteria for inclusion in this study. The data were statistically analyzed with the expectation that this procedure may provide a new feature that could have some clinical significance. Timing delays occurred more frequently than amplitude reductions in the maximal combined response and flicker responses, while amplitude reductions were more common in the first and second oscillatory potentials. The hypernormal b-wave amplitude was rare. The summed amplitude of the oscillatory potentials was highly correlated with the total power of the oscillatory potentials (the frequency domain). A reduction of the second oscillatory potential amplitude was more common than a reduction of the summed amplitude or total power. The electroretinographic component that demonstrates retinal dysfunction in the earlier stage may be a valuable indicator. In the early stage, a delay in the a-wave time and a reduction in the second oscillatory potential amplitude were the most frequent abnormalities: analysis of variance demonstrated that the summed amplitude of the oscillatory potentials and second oscillatory potential amplitude and time were the most sensitive measures of the diabetic retina. Hence, the second oscillatory potential amplitude may be the most sensitive and valuable indicator representing a quantitative measure of overall retinal dysfunction.     

Selective reduction of oscillatory potentials and pattern electroretinograms after retinal ganglion cell damage by disease in humans or by kainic acid toxicity in cats

We recorded pattern electroretinograms, scotopic threshold responses, oscillatory potentials and ganzfeld flash electroretinograms in patients with glaucoma or other optic nerve diseases and in cats with inner retinal damage caused by intravitreal injections of kainic acid. In both studies, the scotopic b-wave and the scotopic threshold responses were normal but the oscillatory potentials and pattern electroretinograms were not. The photopic b-wave was also often reduced in patients with scotopic oscillatory potential reduction, and the reduction was proportionate to the oscillatory potential change. Oscillatory potentials were as frequently reduced as pattern electroretinograms in both patient groups, and in the few cases where only one response was reduced, there was no bias toward either measure. In cats, the effects of intravitreal injection of various doses of kainic acid on the retina were evaluated electrophysiologically, and structural damage was assessed histologically. After 25 nmol of kainic acid, the pattern electroretinograms and oscillatory potentials were reduced but neither the b-waves nor the scotopic threshold responses, were affected. Histologic studies of retinas after this dose showed swollen dendrites that were restricted to the outer part (off-sublamina) of the inner plexiform layer. Serial semithin sections indicated that most, if not all, of the swelling was confined to dendrites of large ganglion cells. Our results indicate that the size and sensitivity of the oscillatory potential response may have a role in the diagnosis and management of early glaucoma and optic nerve disease, and that the photopic electroretinogram may give similar information.     

The pattern electroretinogram (PERG) with contralateral corneal reference

The pattern electroretinogram (PERG) is a sensitive test for investigation of the proximal retina. For monocular recording, the contralateral corneal reference (CCR) electrode position has been suggested as a possible alternative to the conventional ipsilateral temporal reference (ITR). We therefore compared the effect of these electrode positions on 26 subjects (median age 45.5 years, IQR 24.5-61.5). The signals were recorded monocularly with DTL nylon fibre electrodes. The visual stimuli were high contrast 19' black-white checkerboard patterns, reversing 4 times/s. PERG responses obtained from the CCR gave larger amplitudes and the increase in amplitude was statistically significant (P < 0.001). Because the CCR signal is larger, new normative ranges must be established to avoid false negative results.     

Comparisons of the amplitude size and the reproducibility of three different electrodes to record the corneal flash electroretinogram in rodents

To compare corneal electrodes commonly used in rodent eyes for repeat and left versus right eye accuracy and variability to record the flash electroretinogram (ERG). Animals studied were eight C57BL/6 mice and eight rats of the Wistar strain. Scotopic ERGs were recorded from eyes of dark-adapted anesthetized rodents to compare a custom-made gold-wire contact lens electrode (CLE), a cotton-wick silver–silver chloride electrode (SCLE), and a coiled stainless steel wire electrode (SSE). Compared to SCLE and SSE, the potentials recorded by CLE are characterized by significantly larger ERG amplitudes and oscillatory potentials in both rats and mice (p<0.0001). In analyzing test–retest data comparing the three different electrodes the coefficient of variation was smaller (range, 10.3–15.5%) and the interclass correlation coefficient (0.77–0.93) showed a better agreement for the CLE. Recording scotopic ERGs with custom-made gold-wire contact lens electrodes records large amplitudes and shows a good reproducibility and reliability to monitor retinal function in rodent eyes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Improved electrode for electroretinography.

Corneal electrodes useful for clinical electroretinography require topical anesthesia, interfere with vision, can abrade, and are not well accepted by most children and many adults. A low mass conductive thread, corneal (DTL) electrode is described and comparatively tested against the Burian-Allen electrode. The DTL electrode was found to have few of the limitations of the hard contact lens electrode. Furthermore, the DTL electrode signal quality was comparable to that of the Burian-Allen electrode and provided less between-patient variability.