To evaluate the visual function of WT and KO Ndufs4 mice, photopic flash electroretinography (ERG) was performed with Espion ERG Diagnosys machine (Diagnosys, USA). To demonstrate the effect of iPSC-MSC treatment, ERG was performed at 1 and 4 weeks after injection. Before ERG test, mice were anesthetized by i.p. injection of ketamine (100 mg/kg) and xylazine (20 mg/kg), pupils were dilated using with 1% Mydriacyl (Alcon, US). Both eyes were anesthetized topically with 0.5% proparacaine (Alcon, US), ring shaped gold recording electrodes were placed on the cornea of both eyes. A pair of reference needle electrodes made of stainless steel were placed subcutaneously behind the ears and a ground electrode was placed subcutaneously by the tail. After 10 min green light adaption, eyes of the tested mouse were exposed to a series of light flashes in increasing intensities of 11.38 cd.s/m2, 16.60 cd.s/m2 and 22.76 cd.s/m2 with a green background, to record the light response of the retinas. Mice's body temperature was maintained at 37°C by a heating pad during the whole procedure. Results were analyzed for a-wave, b-wave, and PhNR by ERG View v4.380R software.
Erg machine
Manufactured by Diagnosys
Sourced in United States
The ERG Diagnosys machine is a laboratory equipment designed for electroretinography (ERG) testing. It is used to measure the electrical responses of the retina to light stimuli, providing insights into the function of the visual system.
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2 protocols using erg machine
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Photopic ERG Assessment of Ndufs4 Mice
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Electroretinogram Recordings in Mice
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ERGs were recorded with an Espion ERG Diagnosys machine (Diagnosys, Littleton, MA) as previously described by us18 (link). In brief, mice were anaesthetized intraperitoneally with a mixture of Dormitor (1 mg/kg medetomidine hydrochloride; Pfizer, UK) and Ketamine after overnight dark adaption. Pupils were dilated with 1% Mydriacyl (Alcon, Ontario, Canada). Flash ERG was measured using a gold wire corneal electrode, a forehead reference electrode, and a ground electrode near the tail.
Scotopic, rod-mediated responses were obtained from dark-adapted animals at the following increasing light intensities: 0.01 and 3 cd-s/m2. Photopic, cone-mediated responses were performed following 10-minute light adaptation on the background light intensity of 30 cd/m2. Recordings were obtained at the light intensity of 3 cd-s/m2. The ERG a-wave amplitudes were measured from the baseline to the negative peak and the b-wave was measured from the trough of the a-wave to the peak of the positive wave or, when the a-wave was not present, from baseline to the peak of the first positive wave.
Scotopic, rod-mediated responses were obtained from dark-adapted animals at the following increasing light intensities: 0.01 and 3 cd-s/m2. Photopic, cone-mediated responses were performed following 10-minute light adaptation on the background light intensity of 30 cd/m2. Recordings were obtained at the light intensity of 3 cd-s/m2. The ERG a-wave amplitudes were measured from the baseline to the negative peak and the b-wave was measured from the trough of the a-wave to the peak of the positive wave or, when the a-wave was not present, from baseline to the peak of the first positive wave.
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