Yes / Purpose To record transient ERGs from the lightadapted human retina using silent substitution stimuli which selectively reflect the activity of rod photoreceptors. We aim to describe the morphology of these waveforms and examine how they are affected by the use of less selective stimuli and by retinal pathology. Methods Rod-isolating stimuli with square-wave temporal profiles (250/250 ms onset/offset) were presented using a 4 primary LED ganzfeld stimulator. Experiment 1: ERGs were recorded using a rodisolating stimulus (63 ph Td, rod contrast, Crod = 0.25) from a group (n = 20) of normal trichromatic observers. Experiment 2: Rod ERGs were recorded from a group (n = 5) using a rodisolating stimulus (Crod = 0.25) which varied in retinal illuminance from 40 to 10,000 ph Td. Experiment 3: ERGs were elicited using 2 kinds of nonisolating stimuli; (1) broadband and (2) rod-isolating stimuli which contained varying degrees of L- and M-cone excitation. Experiment 4: Rod ERGs were recorded from two patient groups with rod monochromacy (n = 3) and CSNB (type 1; n = 2). Results The rod-isolated ERGs elicited from normal subjects had a waveform with a positive onset component followed by a negative offset. Response amplitude was maximal at retinal illuminances\100 ph Td and was virtually abolished at 400 ph Td. The use of non-selective stimuli altered the ERG waveform eliciting more photopic-like ERG responses. Rod ERGs recorded from rod monochromats had similar features to those recorded from normal trichromats, in contrast to those recorded from participants with CSNB which had an electronegative appearance. Conclusions Our results demonstrate that ERGs elicited by silent substitution stimuli can selectively reflect the operation of rod photoreceptors in the normal, light-adapted human retina. / Deutsche Forschungsgemeinschaft (DFG) (KR1317/13-1) and Bundesministerium für Bildung und Forschung (BMBF) (01DN14009) provided financial support for JK.

Yes / We studied the spatial arrangement of L- and M-cone driven electroretinograms (ERGs) reflecting the activity of magno- and parvocellular pathways. L- and M-cone isolating sine wave stimuli were created with a four primary LED stimulator using triple silent substitution paradigms. Temporal frequencies were 8 and 12 Hz, to reflect cone opponent activity, and 30, 36 and 48 Hz to reflect luminance activity. The responses were measured for full-field stimuli and for different circular and annular stimuli. The ERG data confirm the presence of two different mechanisms at intermediate and high temporal frequencies. The responses measured at high temporal frequencies strongly depended upon spatial stimulus configuration. In the full-field conditions, the L-cone driven responses were substantially larger than the full-field M-cone driven responses and also than the L-cone driven responses with smaller stimuli. The M-cone driven responses at full-field and with 70° diameter stimuli displayed similar amplitudes. The L- and M-cone driven responses measured at 8 and 12 Hz were of similar amplitude and approximately in counter-phase. The amplitudes were constant for most stimulus configurations. The results indicate that, when the ERG reflects luminance activity, it is positively correlated with stimulus size. Beyond 35° retinal eccentricity, the retina mainly contains L-cones. Small stimuli are sufficient to obtain maximal ERGs at low temporal frequencies where the ERGs are also sensitive to cone-opponent processing

Yes / Purpose: To demonstrate that silent substitution stimuli can be used to generate electroretinograms (ERGs) that effectively isolate rod photoreceptor function in humans without the need for dark adaptation, and that this approach constitutes a viable alternative to current clinical standard testing protocols. Methods: Rod-isolating and non-isolating sinusoidal flicker stimuli were generated on a 4 primary light-emitting diode (LED) Ganzfeld stimulator to elicit ERGs from participants with normal and compromised rod function who had not undergone dark-adaptation. Responses were subjected to Fourier analysis, and the amplitude and phase of the fundamental were used to examine temporal frequency and retinal illuminance response characteristics. Results: Electroretinograms elicited by rod-isolating silent substitution stimuli exhibit low-pass temporal frequency response characteristics with an upper response limit of 30 Hz. Responses are optimal between 5 and 8 Hz and between 10 and 100 photopic trolands (Td). There is a significant correlation between the response amplitudes obtained with the silent substitution method and current standard clinical protocols. Analysis of signal-to-noise ratios reveals significant differences between subjects with normal and compromised rod function. Conclusions: Silent substitution provides an effective method for the isolation of human rod photoreceptor function in subjects with normal as well as compromised rod function when stimuli are used within appropriate parameter ranges. Translational Relevance: This method of generating rod-mediated ERGs can be achieved without time-consuming periods of dark adaptation, provides improved isolation of rod- from cone-based activity, and will lead to the development of faster clinical electrophysiologic testing protocols with improved selectivity.

The work in this thesis is concerned with examining the retinal and cortical contributions to human trichromatic colour vision. Chromatic processing at the cortex level was examined using visual evoked potentials (VEPs). These responses were elicited by chromatic spot stimuli, which were manipulated in order to selectively activate the chromatic processing system. Chromatic processing at the retinal level was examined using the electroretinograms (ERGs) for which cone isolating stimuli were used to assess the nature of L and M cone inputs to cone-opponent mechanisms. The results from the VEP experiments suggest VEP morphology is dependent upon 1) chromatic and or luminance contrast content of the stimulus, 2) stimulus size, and 3) extent to which the chromatic stimulus activates either the L/M or S/(L+M) opponent mechanism. The experiments indicate that chromatic stimulation is indexed by large N1 component and small offset responses. Optimal stimulus size for chromatic isolation is 2-4 ° along L/M axes and 6° along S/(L+M) axis. From the ERG experiments, It has been shown that the low (12Hz) and high (30Hz) temporal frequency flickering stimuli can isolate the chromatic and luminance processing mechanisms in the retina. For low temporal frequency ERGs, the L:M ratio was close to unity and L/M phase difference was close to 180°. For high temporal frequency ERGs, the L:M ratio was more than unity and L/M phase difference was close to 90°. In addition to this, the variation in L:M ratio across the retinal eccentricity was also examined. These results suggest, for the chromatic processing, L:M ratio is close to unity independent of retinal eccentricity and individuals. For the luminance processing, L:M ratio is more than unity and depends upon the region of the retina being stimulated. These findings indicate the maintenance of cone selective input for the chromatic processing across the human retina.

The work in this thesis is concerned with examining the retinal and cortical contributions to human trichromatic colour vision. Chromatic processing at the cortex level was examined using visual evoked potentials (VEPs). These responses were elicited by chromatic spot stimuli, which were manipulated in order to selectively activate the chromatic processing system. Chromatic processing at the retinal level was examined using the electroretinograms (ERGs) for which cone isolating stimuli were used to assess the nature of L and M cone inputs to cone-opponent mechanisms. The results from the VEP experiments suggest VEP morphology is dependent upon 1) chromatic and or luminance contrast content of the stimulus, 2) stimulus size, and 3) extent to which the chromatic stimulus activates either the L/M or S/(L+M) opponent mechanism. The experiments indicate that chromatic stimulation is indexed by large N1 component and small offset responses. Optimal stimulus size for chromatic isolation is 2-4 ° along L/M axes and 6° along S/(L+M) axis. From the ERG experiments, It has been shown that the low (12Hz) and high (30Hz) temporal frequency flickering stimuli can isolate the chromatic and luminance processing mechanisms in the retina. For low temporal frequency ERGs, the L:M ratio was close to unity and L/M phase difference was close to 180°. For high temporal frequency ERGs, the L:M ratio was more than unity and L/M phase difference was close to 90°. In addition to this, the variation in L:M ratio across the retinal eccentricity was also examined. These results suggest, for the chromatic processing, L:M ratio is close to unity independent of retinal eccentricity and individuals. For the luminance processing, L:M ratio is more than unity and depends upon the region of the retina being stimulated. These findings indicate the maintenance of cone selective input for the chromatic processing across the human retina.

No / Cone isolating stimuli were used to assess the temporal frequency response characteristics of L- and M-cone electroretinograms (ERGs) in nine trichromatic and four dichromatic human observers. The stimuli comprised sinusoidal temporal modulations varying from 5 to 100 Hz. ERGs were recorded using corneal fiber electrodes and subjected to fast Fourier transform analysis. At low temporal frequencies (<10  Hz<10  Hz) the L- and M-cone ERGs had similar amplitude and exhibited minimal differences in apparent latency. At higher flicker rates (>20  Hz>20  Hz) L-cone ERGs had greater amplitudes and shorter apparent latencies than the M-cone responses. These differences between the L- and M-cone ERGs are consistent with their mediation by chromatic and luminance postreceptoral processing pathways at low and high temporal frequencies, respectively.

No / Electroretinograms (ERGs) elicited by transient, square-wave L- and M-cone isolating stimuli were recorded from human trichromatic (n=19) and dichromatic (n=4) observers. The stimuli were generated on a 4 primary LED stimulator and were equated in terms of cone modulation (cone contrast = 0.11) and retinal illuminance (12,000 trolands). L- and M-cone isolated ERGs had waveforms similar to those observed for luminance responses. However, M-cone ERGs exhibited a phase reversal in their responses to onset and offset stimuli relative to the L-cone responses. This on-off response reversal was observed in trichromats but not dichromats. Simultaneous counter-phase and in-phase combinations of L- and M-cone isolating stimuli generated responses that reflected chromatic and luminance processing, respectively. We conclude that L- and M-cone specific ERGs provide a measure of how photoreceptors contribute to post-receptoral mechanisms.