A fluorescence-based double retrograde tracer strategy for charting central neuronal connections.
Microspheres (beads) tagged with different fluorescent markers can be used for double retrograde axonal tracing of CNS connections. They have several advantages over other double tracer techniques, including ease-of-use, high transport efficiency, distinctive cell labeling and the ability to produce well-defined injection sites. In this protocol we describe the basic procedure for their use, some common problems and how these can be overcome. The protocol, including animal surgery, preparation and delivery of tracer can be completed in approximately 0.5 d. Subsequent histological processing (excluding survival time) can be completed in 0.5-1 d.
Apps
Richard
R
Department of Physiology, School of Medical Sciences, University of Bristol, Bristol, UK. r.apps@bristol.ac.uk
Ruigrok
Tom J H
TJ
D000818:Animals; D001370:Axonal Transport / Q000502:physiology; D002490:Central Nervous System / Q000166:cytology*; D005456:Fluorescent Dyes; D007267:Injections / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D009434:Neural Pathways / Q000166:cytology*; D010477:Perfusion; D051381:Rats
D005456:Fluorescent Dyes