Mice were anesthetized with ketamine/xylazine i.p. and injected i.c.m. into the cisterna magna. 2µl of fluorescent beads mix (0.75µl of FluoSpheres Carboxylate-Modified Microspheres of 1.0µm diameter (ThermoFisher); 0.5µl of FluoSpheres Carboxylate-Modified Microspheres 0.5µm diameter (ThermoFisher) in a total volume of 2µl PBS), 5µl of FluoSpheres Carboxylate-Modified Microsphere of 0.5µm, or 5µl of Alexa-fluor 594/647 conjugated OVA (0.5mg/ml; ThermoFisher) at a rate of 0.5 or 3µl/min. After injection, the needle is left in place for 1–2 minutes to avoid backflow. Subsequently, the muscle was pinched to apply some pressure on the cisterna magna prior to the needle removal. Mice were then sutured and allowed to recover on a heating pad until responsive (after about 45 min to an hour). Lymph nodes and meninges were harvested at the indicated time points and analyzed by immunohistochemistry or flow cytometry.
Fluospheres carboxylate modified microspheres 0.5 m diameter
Manufactured by Thermo Fisher Scientific
FluoSpheres Carboxylate-Modified Microspheres 0.5µm diameter are fluorescent polystyrene particles with carboxylate surface modifications. They are designed for use as calibration standards, probes, and labels in various analytical and biomedical applications.
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2 protocols using fluospheres carboxylate modified microspheres 0.5 m diameter
1
Intracisternal Injection of Fluorescent Tracers
2
Intracisternal Injection of Fluorescent Tracers
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Mice were anesthetized with ketamine/xylazine i.p. and injected i.c.m. into the cisterna magna. 2µl of fluorescent beads mix (0.75µl of FluoSpheres Carboxylate-Modified Microspheres of 1.0µm diameter (ThermoFisher); 0.5µl of FluoSpheres Carboxylate-Modified Microspheres 0.5µm diameter (ThermoFisher) in a total volume of 2µl PBS), 5µl of FluoSpheres Carboxylate-Modified Microsphere of 0.5µm, or 5µl of Alexa-fluor 594/647 conjugated OVA (0.5mg/ml; ThermoFisher) at a rate of 0.5 or 3µl/min. After injection, the needle is left in place for 1–2 minutes to avoid backflow. Subsequently, the muscle was pinched to apply some pressure on the cisterna magna prior to the needle removal. Mice were then sutured and allowed to recover on a heating pad until responsive (after about 45 min to an hour). Lymph nodes and meninges were harvested at the indicated time points and analyzed by immunohistochemistry or flow cytometry.
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