Thy1-YFP and Thy1-YFP:5XFAD mice were injected with Alexa594-conjugated fibrinogen (Invitrogen) intravenously daily for 3 days as described15 (link). Mice were also injected i.p. with Methoxy-X04 24 h prior to imaging. Methoxy-X04 was solubilized with DMSO:propylene glycol:PBS pH 7.5 (ratio 2:9:9) at 5 mg/ml. On day of imaging, a small craniotomy was made and a custom-made metal plate affixed to a stage to stabilize the skull. Alexa594-fibrinogen solution was injected retro-orbitally prior to imaging. The anesthetized animal was placed on a heated pad under an Ultima-IV multiphoton microscope (Prairie) equipped with MaiTai DeepSee-eHP lasers (Spectra Physics). The excitation wavelength was 820nm to simultaneously visualize fibrinogen, methoxy-X04 and YFP dendrites. Imaging was performed from 20 to 150 μm below the dura, using a Nikon 40 × 0.8 NA -immersion lenses with a 1.0 μm z-step. z-stacks of images were projected along the z-axis to recreate two-dimensional representations of the 3D structures within the imaged volumes. Images were adjusted for brightness, contrast and background noise with ImageJ. Spectral unmixing plugin in ImageJ was used to separate overlapping signals.
40 0.8 na immersion lenses
Manufactured by Nikon
The Nikon 40 × 0.8 NA immersion lenses are high-performance microscope objective lenses designed for laboratory applications. They provide a numerical aperture of 0.8, which enables high-resolution imaging. These lenses are optimized for use with immersion oil to enhance optical performance.
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2 protocols using 40 0.8 na immersion lenses
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Imaging Alzheimer's Disease Pathology
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Imaging Alzheimer's Disease Pathology
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Thy1-YFP and Thy1-YFP:5XFAD mice were injected with Alexa594-conjugated fibrinogen (Invitrogen) intravenously daily for 3 days as described15 (link). Mice were also injected i.p. with Methoxy-X04 24 h prior to imaging. Methoxy-X04 was solubilized with DMSO:propylene glycol:PBS pH 7.5 (ratio 2:9:9) at 5 mg/ml. On day of imaging, a small craniotomy was made and a custom-made metal plate affixed to a stage to stabilize the skull. Alexa594-fibrinogen solution was injected retro-orbitally prior to imaging. The anesthetized animal was placed on a heated pad under an Ultima-IV multiphoton microscope (Prairie) equipped with MaiTai DeepSee-eHP lasers (Spectra Physics). The excitation wavelength was 820nm to simultaneously visualize fibrinogen, methoxy-X04 and YFP dendrites. Imaging was performed from 20 to 150 μm below the dura, using a Nikon 40 × 0.8 NA -immersion lenses with a 1.0 μm z-step. z-stacks of images were projected along the z-axis to recreate two-dimensional representations of the 3D structures within the imaged volumes. Images were adjusted for brightness, contrast and background noise with ImageJ. Spectral unmixing plugin in ImageJ was used to separate overlapping signals.
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