The visual acuity and contrast sensitivity were measured by OKN using the OptoMotry system of Cerebral Mechanics (Lethbridge, AB, Canada). Both visual acuity and contrast sensitivity were measured once before injury and again at 14 DPI and at 35 DPI. The visual acuity was assessed at 100% contrast by varying the spatial frequency threshold while the contrast sensitivity was measured at a spatial frequency of 0.042 cycles per degree (c/d). Six mice of both genders were used for each treatment group (NM and saline). The visual acuity and contrast sensitivity data represented for each mouse were the average of the data from both the eyes of the individual mice.
Optomotry system
Manufactured by Cerebral Mechanics
Sourced in Canada, United States
The OptoMotry system is a laboratory equipment used for visual function assessment. It measures optokinetic responses in animals to evaluate visual capabilities. The system consists of a virtual reality arena that generates moving visual stimuli, allowing researchers to assess the animal's visual behavior and motor responses.
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Lab products found in correlation
C57bl 6j mice, by Jackson ImmunoResearch (3 mentions)
Optomotry, by Cerebral Mechanics (2 mentions)
Dcr hc26, by Sony (2 mentions)
Micron 4 retinal imaging microscope, by Phoenix Pharmaceuticals (1 mentions)
2 2 2 tribromoethanol, by Merck Group (1 mentions)
Prism 7, by GraphPad (1 mentions)
Video camera, by Zeiss (1 mentions)
Optomotry software, by Cerebral Mechanics (1 mentions)
50 protocols using optomotry system
1
Visual Acuity and Contrast Sensitivity
2
Measuring Visual Acuity and Contrast Sensitivity
3
Assessing Visual Acuity in Mouse Models
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The photopic optomotor response of mice was measured using the OptoMotry System (CerebralMechanics) at a background light of ≈70 cd/m2 as previously described (Xiong et al., 2019 (link)). The contrast of the grates was set to be 100%, and temporal frequency was 1.5 Hz. The threshold of mouse visual acuity (i.e., maximal spatial frequency) was tested by an examiner without knowledge of the control vs. experimental groups. During each test, the direction of movement of the grates (i.e., clockwise or counterclockwise) was randomized, and the spatial frequency of each testing episode was determined by the software. Without knowing the spatial frequency of the moving grates, the examiner reported either ‘yes’ or ‘no’ to the system until the threshold of acuity was determined by the software. Optomotor tests were conducted on rd10 and Rho-/- mice, but not with rd1 strain, which loses vision at a very early age, before any meaningful test could be performed.
4
Visual Function Assessment in Crb1/Crb2 Mice
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Spatial frequency and contrast sensitivity thresholds were measured using an OptoMotry system (Cerebral Mechanics, Lethbridge, AB, Canada). One-, 3-, 7-, and 9-month-old Crb1KOCrb2ΔRods and Crb1KOCrb2flox/flox mice, and 9-month-old Crb2flox/flox mice were placed on a small platform in the center of four computer monitors that formed a virtual drum with a rotating vertical sine wave grating (12°/s (d/s)), as described previously [29 (link)]. Reflexive head movements in the same direction as the rotating gratings were considered positive responses. Spatial frequency thresholds were determined with an increasing staircase paradigm starting at 0.042 cycles/deg (c/d) with 100% contrast. Contrast sensitivity thresholds were measured across six spatial frequencies (0.031, 0.064, 0.092, 0.103, 0.192, and 0.272 c/d). The reciprocal of the contrast sensitivity threshold was used as the contrast sensitivity value at each spatial frequency.
The numbers of mice used for the OKT measurements per time point were as follows: 1M (Crb1KOCrb2flox/flox = 8 and Crb1KOCrb2ΔRods = 4), 3M (Crb1KOCrb2flox/flox = 10 and Crb1KOCrb2ΔRods = 10), 5M (Crb2flox/flox = 10, Crb2ΔRods = 8), 7M (Crb1KOCrb2flox/flox = 8 and Crb1KOCrb2ΔRods = 9), 9M (Crb2flox/flox = 7, Crb1KOCrb2flox/flox = 8, and Crb1KOCrb2ΔRods = 9). Statistical significance was calculated by using Mann-Whitney U test.
The numbers of mice used for the OKT measurements per time point were as follows: 1M (Crb1KOCrb2flox/flox = 8 and Crb1KOCrb2ΔRods = 4), 3M (Crb1KOCrb2flox/flox = 10 and Crb1KOCrb2ΔRods = 10), 5M (Crb2flox/flox = 10, Crb2ΔRods = 8), 7M (Crb1KOCrb2flox/flox = 8 and Crb1KOCrb2ΔRods = 9), 9M (Crb2flox/flox = 7, Crb1KOCrb2flox/flox = 8, and Crb1KOCrb2ΔRods = 9). Statistical significance was calculated by using Mann-Whitney U test.
5
Electrophysiological and Visual Acuity Assessments in Mice
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ERG measurements were done as described previously (Kim et al., 2017 (link)). In brief, mice were either dark- or light-adapted for 12 h and anesthetized with 2,2,2-tribromoethanol (Sigma, USA) in prior to dilating the pupils of the mice by 0.5% tropicamide. The mice were placed with a gold-plated objective lens on their corneas and silver-embedded needle electrodes at their foreheads and tails. The ERG recordings were performed using Micron IV retinal imaging microscope (Phoenix Research Labs, USA) and analyzed by Labscribe ERG software according to the manufacturer’s instruction.
Mouse visual acuity was measured with the OptoMotry system (Cerebral Mechanics, USA) as previously described (Prusky et al., 2004 (link)). Mice adapted to ambient light for 30 min were placed on the stimulus platform surrounded by four computer monitors displaying black and white vertical stripe patterns. An event that mice track the stripe movements with reflexive head-turn was counted as a successful visual detection. The detection thresholds were then obtained from the OptoMotry software.
Mouse visual acuity was measured with the OptoMotry system (Cerebral Mechanics, USA) as previously described (Prusky et al., 2004 (link)). Mice adapted to ambient light for 30 min were placed on the stimulus platform surrounded by four computer monitors displaying black and white vertical stripe patterns. An event that mice track the stripe movements with reflexive head-turn was counted as a successful visual detection. The detection thresholds were then obtained from the OptoMotry software.
6
Measuring Optokinetic Response in Mice
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The optokinetic response was measured using an OptoMotry system according to previously described methods (Cerebral Mechanics, Lethbridge, Alberta, Canada) [11] (link). Briefly, mice were placed onto an elevated platform in the test chamber, and tested after ∼60 seconds of brief acclimation. Mice were presented with a vertically oriented sine wave grating rotating at 12°/s. The between stimulus ‘blank’ was an equal luminance gray homogenous surround (152 cd.sm2). The level of contrast was fixed at 100%, and the spatial frequency was varied to determine thresholds for stimulus detection using a standard protocol. Spatial frequency was expressed in cycles per degree (c/d) for one sine wave (paired vertical black and white bars). Tracking head movements were scored by an experienced operator blind to genotype. Test sessions were during the mid 4 hours of the light phase of a daily cycle. The test chamber was thoroughly cleaned between animals. Where no response was observed, an animal was tested at least 10 times.
7
Evaluating Visual Acuity and Contrast Sensitivity in Rats
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Assessment of visual function was performed using an OptoMotry system (OptoMotry™, CerebralMechanics, Lethbridge, AB, Canada) [23 (link)]. This system was used to evaluate the visual acuity (VA) and contrast sensitivity (CS) of both P23HxLE and wild-type rats monthly from postnatal day 30 (P30) to P180. The maximum VA and minimum contrast identified were reached when the rat could not distinguish the stimuli presented and were judged by the experimenter using a video camera, which provides real-time feedback on a computer. VA and CS results depend on the experience and training of the experimenter, and thus all the rat-tracking movements were assessed by the same researcher.
8
Measuring Visual Acuity in Mice
9
Measuring Visual Function in Mice
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Visual function was measured with optomotor response (OMR) testing at 5, 9, 13, 17, and 21 weeks of age, as previously described37 (link)–39 using the OptoMotry system (Cerebral Mechanics, USA).40 (link) Briefly, a mouse was placed on a central pedestal in an enclosed chamber of monitors, which display rotating vertical sine wave gratings of varying spatial frequency (with contrast set at 100%) or varying grating contrast (with spatial frequency set at 0.103 cycles/degree). A trained observer marked when the characteristic head tracking movement occurred (or did not occur) through a programmed staircase method to calculate the visual threshold. Measurements from the left and right eye were averaged for a combined visual threshold score. Contrast threshold results were converted to Michelson contrast values.41 (link)
10
Optokinetic Tracking for Visual Acuity
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Visual acuity was evaluated in rd10-nontreated, rd10+MMF mice, and rd10+PTZ mice ages P21, P28 and P41; a cohort of wild-type (WT) C57Bl/6J mice (Jackson Laboratories) were also included in the assessment for comparison. Acuity was measured as described previously30 (link) following the method of Prusky et al.31 (link) Spatial thresholds for optokinetic tracking of sine-wave gratings were measured using the OptoMotry system (CerebralMechanics, Medicine Hat, Alberta, Canada). Mice were placed unrestrained on a pedestal and were presented vertical sine-wave gratings moving at 12°/s or gray of the same mean luminance within the OptoMotry device, which functions as a virtual cylinder. The cylinder hub was continually centered between the mouse's eyes to establish the spatial frequency of the grating at the mouse's viewing position as it shifted its position. Gray color was projected while the mouse was moving; when movement ceased, the gray was replaced with the grating. Grating rotation under these circumstances elicited reflexive tracking, which was scored via live video using a method of limits procedure with a yes/no criterion as recommended by the manufacturer. A measure of spatial resolution was taken as the asymptote of a staircase procedure. The two eyes were tested in an interleaved fashion.
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