Preparation of ultrathin sections of retinal tissue for examination by TEM has been described previously.13 (link),14 (link) Briefly, a 2 × 2 mm piece of retinal tissue was removed 5 mm above the superior margin of the optic disc. Retinal tissue was rinsed in buffer, postfixed in 2% osmium tetroxide and 1.5% potassium ferrocyanide in dH2O for 2 hours, dehydrated in a graded series of ethanols and embedded in Epon-Araldite (Electron Microscopy Sciences, Hatfield, PA). Ultrathin sections (90 nm) of embedded retinal tissue were cut from each block with a diamond knife (Micro Star Technologies, Inc., Huntsville, TX) using an ultramicrotome (Ultracut E 701704; Reichert-Jung, Buffalo, NY). Ultrathin sections were collected on copper grids, and counterstained with 4% methanolic uranyl acetate (Electron Microscopy Sciences). Photoreceptor structure was then examined with a transmission electron microscope (Model 300: Phillips, Eindhoven, The Netherlands).
Model 300
Manufactured by Philips
Sourced in Netherlands
The Philips Model 300 is a versatile lab equipment designed for general laboratory applications. It features a compact and durable construction. The core function of this model is to provide reliable and consistent performance in various laboratory settings.
Automatically generated - may contain errors
4 protocols using model 300
1
Retinal Tissue Ultrastructural Analysis
2
Outer Retinal Morphology Examination Protocol
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Sections for EM were prepared as previously described68 (link). Briefly, hemisected eyecups were rinsed in buffer and postfixed in 2% osmium tetroxide and 1.5% potassium ferrocyanide in dH2O for 2 h, dehydrated in a graded ethanol series, and embedded in Epon-Araldite (Electron Microscopy Sciences, Hatfield, PA). Semi-thin sections (4 μm) were cut and stained with 1% cresyl violet. Ultra-thin sections (90 nm) were cut on an ultramicrotome (Ultracut E 701704, Reichert-Jung, Buffalo, NY) using a diamond knife (Micro Star Technologies, Inc., Huntsville, TX), collected on copper grids, counterstained with 4% methanolic uranyl acetate (Electron Microscopy Sciences, Hatfield, PA), and outer retinal morphology examined using a transmission electron microscope (TEM; Model 300: Phillips, Eindhoven, The Netherlands). Photomicrographs were captured with a digital camera (15-megapixel digital camera, Scientific Instruments and Applications, Duluth, GA) and Maxim DL Version 5 software (Diffraction Limited, Ottawa, Canada).
3
Retinal Ultrastructure Visualization via TEM
4
Transmission Electron Microscopy of Retinal Tissue
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Sections for EM were prepared as previously described 46 (link) . Briefly, hemisected eyecups were rinsed in buffer and postfixed in 2% osmium tetroxide and 1.5% potassium ferrocyanide in dH2O for 2 hr, dehydrated in a graded ethanol series, and embedded in Epon-Araldite (Electron Microscopy Sciences, Hatfield, PA). Semi-thin sections (4 μm) were cut and stained with 1% cresyl violet. Ultra-thin sections (90 nm) were cut on an ultramicrotome (Ultracut E 701704, Reichert-Jung, Buffalo, NY) using a diamond knife (Micro Star Technologies, Inc., Huntsville, TX), collected on copper grids, counterstained with 4% methanolic uranyl acetate (Electron Microscopy Sciences, Hatfield, PA), and outer retinal morphology examined using a transmission electron microscope (TEM; Model 300: Phillips, Eindhoven, The Netherlands).
Photomicrographs were captured with a digital camera (15-megapixel digital camera, Scientific Instruments and Applications, Duluth, GA) and Maxim DL Version 5 software (Diffraction Limited, Ottawa, Canada).
Photomicrographs were captured with a digital camera (15-megapixel digital camera, Scientific Instruments and Applications, Duluth, GA) and Maxim DL Version 5 software (Diffraction Limited, Ottawa, Canada).
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