Multiple axonal tracing: simultaneous detection of three tracers in the same section
Palabras clave : 
Tract-tracing
PHA-L
BDA
Fluoro-Gold
V-VIP
Peroxidase
Fecha de publicación: 
1998
Editorial : 
Springer Verlag
ISSN: 
1432-119X
Cita: 
Lanciego JL, Wouterlood FG, Erro E, Gimenez-Amaya JM. Multiple axonal tracing: simultaneous detection of three tracers in the same section. Histochem Cell Biol 1998 Nov;110(5):509-515.
Resumen
Multiple neuroanatomical tract-tracing methods are important tools for elucidating the connectivity between different populations of neurons. Evaluation of the question as to whether two specific fiber inputs converge on a particular, identified population of projection neurons requires the application of a triple-staining procedure that allows the unequivocal detection of three markers in a single section. The present report deals with a combination of tracing methods using anterogradely transported Phaseolus vulgaris leucoagglutinin and biotinylated dextran amine in conjunction with retrogradely transported Fluoro-Gold. These tracers were simultaneously detected according to a three-color paradigm, which includes the use of three different peroxidase substrates (nickel-enhanced diaminobenzidine, diaminobenzidine, and Vector VIP), thus resulting in three distinct precipitates: black, brown, and purple. We illustrate this method by showing convergence of projections arising from neurons located in two separate basal ganglia-related nuclei onto identified thalamostriatal projection neurons.

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