After anesthesia and pupil dilation, the retinal fundus was captured with the Micron IV fundus camera with mouse objective and 50-degree field of view (1.8-mm diameter) centering on the optic cup (Phoenix Research Labs, Pleasanton, CA, USA). Retinal infiltrates and folds were counted in the resulting digital images. The grading system were based on the retinal structural damage scores from 0 to 4. Score 1 refers to 1–4 dot-like lesion or 1 linear lesion, score 2 refers to 5–10 dot-like lesions or 2–3 linear lesions, score 3 refers to >10 dot-like lesions or >3 linear lesions, score 4 refers to confluent linear lesions [23 (link), 61 (link)].
Micron 4 fundus camera
Manufactured by Phoenix Pharmaceuticals
Sourced in United States
The Micron IV fundus camera is a specialized laboratory equipment designed for high-resolution imaging of the fundus, the interior surface of the eye. It captures detailed images of the optic nerve, retina, and blood vessels within the eye.
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Lab products found in correlation
Spectralis hra oct device, by Heidelberg Engineering (2 mentions)
Micron 4 retinal imaging microscope, by Phoenix Pharmaceuticals (1 mentions)
Micron 4, by Phoenix Pharmaceuticals (1 mentions)
Ab14196, by Abcam (1 mentions)
Fluorescein solution, by Alcon (1 mentions)
Mydrum, by Bausch & Lomb (1 mentions)
Irbp 161 180 peptide, by AnaSpec (1 mentions)
B10 riii mice, by Jackson ImmunoResearch (1 mentions)
Complete freund s adjuvant, by Merck Group (1 mentions)
Fluorescite 10, by Alcon (1 mentions)
Envisu r2210, by Leica (1 mentions)
Micron oct, by Phoenix Pharmaceuticals (1 mentions)
Streampix 6, by Phoenix Pharmaceuticals (1 mentions)
Oct scan head, by Phoenix Pharmaceuticals (1 mentions)
Tropicamide, by Santen (1 mentions)
Isoflurane, by Kent Scientific (1 mentions)
Isoflurane vaporizer, by Kent Scientific (1 mentions)
10 protocols using micron 4 fundus camera
1
Retinal Fundus Imaging and Structural Damage
2
Laser-Induced Retinal Damage in Mice
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The mice were placed on the imaging platform Phoenix Micron IV retinal imaging microscope after being anesthetized and prepared, as mentioned in the FA and OCT imaging. The fundus was viewed with the Micron IV fundus camera, and laser photocoagulation was induced using the image-guided laser system (Micron IV, Phoenix Research Laboratories, Pleasanton, CA, USA). The fundus image as well as the aiming beam could be observed on the monitor screen. Four–five laser burns at an equal distance from the optic nerve were induced one by one in each eye by a green Argon laser pulse, with a wavelength of 532 nm, a duration of 70 ms, and power levels from 250 mW to 260 mW. Successful laser burns were confirmed by the appearance of white bleep with grey outline, indicating the break of Bruch’s Membrane. After laser photocoagulation, the mice were then placed under an infra-red warming lamp until they awakened.
3
In-vivo Analysis of Retinal Degeneration
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On days 7 and 30, fundus photography and OCT images were taken using micron IV fundus camera (Phoenix Research Laboratories, USA) for both Ngb and sham treated eyes. Clinical signs of vitritis, retinitis and optic neuropathy were examined by an experienced clinician. OCT images were analyzed using InSight software V. 1.1 to determine retinal thickness and sign of retinal degeneration. Electroretinogram (ERG) was recorded after overnight dark-adaptation (>12 hours) using corneal monopolar electrodes and an Espion system (Espion, Diagnosys LLC, USA). The stimulus intensity ranging from −3.0 to 1.0 log cd.s.m−2 was used. The harvested retinae were used to analyze the expression of the chemokines, and histology. The eyes were fixed in 10% neutral buffered formalin immediately after enucleation and embedded in paraffin blocks for histology. These were sectioned into four-micron sections and stained with Hematoxylin and Eosin staining (H&E) for morphology, retinal thickness and features of inflammation (presence of macrophages and lymphocytes). Annexin V immunohistochemistry (abcam, ab14196, 1:200 dilution), a marker for apoptosis, was performed to detect apoptotic cells.
4
Comprehensive Multimodal Retinal Imaging in Mice
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Mice were anesthetized as described above and pupils dilated with 5 mg/mL tropicamide (Mydrum; Bausch + Lomb) and phenylephrine (Neosynephrin-POS 10%; Ursapharm). Optical coherence tomography (OCT) analysis was performed with a Bioptigen Envisu R2210 device (Leica Microsystems) equipped with a lens specifically designed for mouse retinae (50 degrees field of view). Infrared Reflectance (IR) and Blue Autofluorescence (BAF) images were recorded with a Spectralis HRA/OCT device (Heidelberg Engineering) using a 55 degrees widefield lens. For fundus fluorescein angiography (FFA), 200 µL of a 0.2 % fluorescein solution (Alcon) was injected subcutaneously and pictures were recorded 90 seconds after injection with the Spectralis HRA/OCT device (Heidelberg Engineering). During image acquisition, mouse contact lenses (Heidelberg Engineering) were applied to the mouse eyes to prevent drying and improve the image quality. Fundus pictures were recorded with a Micron IV Fundus camera (Phoenix) equipped with a mouse-specific objective.
5
Rat Pupil Dilation and Fundus Imaging
6
IRBP-Induced Experimental Autoimmune Uveitis in Mice
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Female 8- to 12-week-old B10.RIII mice were purchased from Jackson Laboratory (Sacramento, CA, USA) or bred at Oregon Health & Science University (OHSU). All mice were maintained in accordance with the institutional policies for animal health and well-being, in ventilated cages under HEPA-filtered barrier conditions, and fed gamma-irradiated food and water ad libitum. Mice were immunized with an emulsion containing 15 μg interphotoreceptor retinoid binding protein (IRBP)161–180 peptide (Anaspec, Fremont, CA, USA) in complete Freund's adjuvant (Sigma-Aldrich Corp., St. Louis, MO, USA) containing 2.5 mg/mL killed Mycobacterium tuberculosis (MTB) antigen (Difco, Houston, TX, USA)14 (link),15 (link) or with emulsion containing only MTB antigen as an adjuvant-only control. A scrambled IRBP peptide (scIRBP) (Genscript, Township, NJ, USA) was used as an irrelevant antigen control, administered in an emulsion with the same amount of MTB. Clinical EAU score was evaluated by fundus examination using a 90 diopter (D) lens (Volk, Mentor, OH, USA; Keeler, Sacramento, CA, USA) every 3 days. Clinical and histopathologic scoring of uveitis was conducted based on a grading scale of 0 to 4 as described previously.15 (link) Mouse fundus photos were obtained using a Micron IV fundus camera (Phoenix Research Labs, Pleasanton, CA, USA).
7
Rodent Retinal Imaging and Fluorescein Angiography
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The cornea was anesthetized with topical Alcaine (0.5%) and the pupils were dilated with atropine (1%) and phenylephrine (10%). Methylcellulose eye gel was applied to prevent the dehydration of the cornea during imaging. The retinal fundi of the rat were viewed and photographed with a Micron IV fundus camera (Phoenix Research Labs, Inc., Pleasanton, CA). All images were collected by using the specialty Micron IV software (StreamPix; NorPix, Inc., Montreal, Quebec, Canada). Following a subcutaneous injection of 0.1 mL 2.0% fluorescein sodium solution (Fluorescite 10%; Alcon), fluorescein angiographs were viewed and captured with a green filter attached to the Micron III fundus camera at 5 minutes post injection. Images were exported as tagged image files (.tif) and fluorescence intensity was quantified by using a custom macro in ImageJ v1.47.
8
Pupil Dilation and Ocular Imaging in Mice
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In brief, mice were anesthetized, and pupils were dilated with 5 mg/mL tropicamide (Mydrum; Bausch + Lomb, Rochester, NY, USA) and phenylephrine (Neosynephrin-POS 10%; Ursapharm, New Delhi, India). Optical coherence tomography (OCT) recordings were done with a Bioptigen Envisu R2210 device (Leica Microsystems, Wetzlar, Germany) equipped with a lens designed for mouse eyes (50° field of view). Fundus pictures were recorded with a Micron IV Fundus camera (Phoenix Research Laboratories, Pleasanton, CA, USA). Pictures of mouse eyes (pupil size) were taken with a Spectralis HRA/OCT device (Heidelberg Engineering, Heidelberg, Germany) approximately 20 minutes after pupil dilation as described above. The same eyes were imaged three or four times: Before AAV injection (pretreatment value) and one week (AAV-stuffer control only) and three and six weeks after AAV injection.
9
Retinal Thickness Monitoring via OCT Imaging in Mouse Laser Injury Model
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OCT images were taken 1, 3, 7, 14, 30, 90, and 180 days after laser photocoagulation, as were corresponding OCT scans using a Micron IV fundus camera and an OCT Scan Head equipped with a mouse objective lens (Phoenix Research Labs, Pleasanton, CA, USA). The OCT device featured a broadband superluminescent diode at 830 nm, customized for retinal imaging of mice. The scan region on the mouse retina was 1.8 mm in the X and Y directions. Linear OCT scans consisted of a series of 1024 single point A-Scans. Right eyes had previously been dilated with 0.5% tropicamide (Santen Pharmaceutical Co. Ltd., Osaka, Japan) and hydroxyl ethyl cellulose (Senju Pharmaceutical Co. Ltd., Osaka, Japan), used as coupling gel. Images were captured from 20 positions for each eye using StreamPix 6 and Micron OCT commercial software (Phoenix Research Labs). Captured images were quantitatively analysed using “In Sight” software, which can automatically detect and measure each retinal layer. Using this software, the retinal thickness was measured at 20 recorded positions for each eye, and the average of all positions represented the overall retinal thickness.
10
Fundus Imaging of Dilated Mice
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Fundus examination was performed as previously described [42 (link)], using a Micron IV fundus camera (Phoenix Research Laboratories, Pleasanton, CA, USA), with the exception that 1% cyclopentolate or 1% atropine was used as the dilating agent, and mice were anesthetized with isoflurane (isoflurane vaporizer from Kent Scientific, Torrington, CT, USA) for the duration of imaging.
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