To visualize vascular dynamics in the mouse skin, we used in vivo multiphoton microscopy, which was a modification of conventional single photon methods69 (link). Mice were anesthetized by injection with urethane (1.5 g/kg), and they were secured to the heated stage (Tokai Hit, Fujinomiya, Japan) of an inverted microscope (Nikon, Eclipse Ti, Tokyo, Japan). Texas-Red-dextran (D1830, 25 mg/kg BW, 70 kDa, Life technologies) and Hoechst 33342 (H1399, 10 mg/kg, Life technologies) were injected into the mice to visualize cell dynamics and blood flow. The tissue was excited at a wavelength of 860 nm using a Ti: sapphire laser (Visio II, Coherent, Santa Clara, CA, USA), and images were captured using a Nikon A1R-MP system as XYZ-T images. Z stacks were usually approximately 50 μm thick with 1-μm slice images. A 40x (N.A. 1.15) water immersion objective lens (Nikon) was used, and the images were captured at 1.5x zoom. Some sequential images were collected for 1 min at a frame rate of 30 XY images/sec. More than five animals were examined in each group. Images were analyzed using NIS-Elements software (Nikon).
Texas red dextran d1830
Manufactured by Thermo Fisher Scientific
Texas-Red-dextran (D1830) is a fluorescent dye-labeled dextran molecule produced by Thermo Fisher Scientific. It is designed for use as a tracer or marker in various biological applications. The core function of Texas-Red-dextran is to provide a fluorescent signal that can be detected and visualized using appropriate imaging techniques.
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2 protocols using texas red dextran d1830
1
Visualizing Vascular Dynamics in Mouse Skin
2
Visualizing Vascular Dynamics in Mouse Skin
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To visualize vascular dynamics in the mouse skin, we used in vivo multiphoton microscopy, which was a modification of conventional single photon methods69 (link). Mice were anesthetized by injection with urethane (1.5 g/kg), and they were secured to the heated stage (Tokai Hit, Fujinomiya, Japan) of an inverted microscope (Nikon, Eclipse Ti, Tokyo, Japan). Texas-Red-dextran (D1830, 25 mg/kg BW, 70 kDa, Life technologies) and Hoechst 33342 (H1399, 10 mg/kg, Life technologies) were injected into the mice to visualize cell dynamics and blood flow. The tissue was excited at a wavelength of 860 nm using a Ti: sapphire laser (Visio II, Coherent, Santa Clara, CA, USA), and images were captured using a Nikon A1R-MP system as XYZ-T images. Z stacks were usually approximately 50 μm thick with 1-μm slice images. A 40x (N.A. 1.15) water immersion objective lens (Nikon) was used, and the images were captured at 1.5x zoom. Some sequential images were collected for 1 min at a frame rate of 30 XY images/sec. More than five animals were examined in each group. Images were analyzed using NIS-Elements software (Nikon).
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