ABSTRACT
This chapter introduces the art of double labeling recorded neurons with the combined use of an intracellular labeling technique and an immunocytochemical procedure. For experiments combining intracellular recording and labeling, an animal is decapitated and the brain gently removed from the skull. The intracellularly labeled neuron is the distinctly stained element in the tissue and can be easily reconstructed with serial sections for a complete morphological analysis. The only disadvantage of biocytin as an intracellular tracer is that a detergent is required to enhance the penetration of probes conjugated to avidin into the tissue sections to bind to the biocytin. The characteristics of the microelectrode are determined by the need for acceptable intracellular recordings rather than by any special requirements for injection of biocytin. The red fluorescing neuron injected with biocytin was photographed. The patch pipette contained both Lucifer Yellow and biocytin, which were slowly diffused into the cell during recording.
