The VSV-GFP is a recombinant virus expressing the green fluorescent protein. The Cy3 fluorescein was modified on the RVG to provide a red fluorescent signal. In the cellular model, the virus-infected N2a cells (MOI of 1) were incubated with 100 μL of T-705@MSN-RVG (2 mg/mL) at 37 °C for 1h. Then, cells were washed three times and stained by a Hoechst (Thermo Fisher, Waltham, MA, USA) at 37 °C for 5 min. Subsequently, the cells were visualized by a TS100 fluorescence microscope (Nikon, Tokyo, Japan).
In the mouse infection model, co-localization was determined in the mouse brain. Briefly, six-week-old female Balb/c mice were infected with VSV-GFP at 200 FFU or DMEM by intracranial (i.c.) route. Then, the mice were inoculated with 25 μL of T-705@MSN-RVG (1 mg/mL) by i.v. injection. The mouse brains were collected at 2 dpi and fixed in 4% paraformaldehyde for 48 h, followed by being dehydrated in 30% sucrose solution and embedded in SAKURA Tissue-Tek® O.C.T compound (SAKURA, Torrance, CA, USA) for rapid freezing and slicing [35 (link)]. The slices were stained by Hoechst (Thermo Fisher, Waltham, MA, USA) at 37 °C for 5 min and then visualized by a TS100 fluorescence microscope (Nikon, Tokyo, Japan).
In the mouse infection model, co-localization was determined in the mouse brain. Briefly, six-week-old female Balb/c mice were infected with VSV-GFP at 200 FFU or DMEM by intracranial (i.c.) route. Then, the mice were inoculated with 25 μL of T-705@MSN-RVG (1 mg/mL) by i.v. injection. The mouse brains were collected at 2 dpi and fixed in 4% paraformaldehyde for 48 h, followed by being dehydrated in 30% sucrose solution and embedded in SAKURA Tissue-Tek® O.C.T compound (SAKURA, Torrance, CA, USA) for rapid freezing and slicing [35 (link)]. The slices were stained by Hoechst (Thermo Fisher, Waltham, MA, USA) at 37 °C for 5 min and then visualized by a TS100 fluorescence microscope (Nikon, Tokyo, Japan).
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