Intracellular filling and reconstruction of identified neurons in fixed rat brain slices.
A simple technique for the morphological characterization of pre-labelled neurons in fixed brain slices is described. Neurons are retrogradely-labeled with a carbocyanine dye and the tissue is fixed and sliced. Individual labeled cells from the interior of a slice are then visualized on an upright fluorescence microscope and impaled with a micropipet containing rhodamine or fluorescein-peroxidase conjugates. The cells are filled by iontophoresis, postfixed, and the peroxidase is oxidized into a permanent, opaque reaction product. The dendritic morphology of the neurons is then reconstructed under camera lucida and quantified.
Prusky
G
G
Department of Psychology and Neuroscience, University of Lethbridge, Alberta, Canada. prusky@uleth.ca
Arjannikova
T
T
D015100:3,3'-Diaminobenzidine / Q000032:analysis; D000818:Animals; D001921:Brain / Q000166:cytology*; D005260:Female; D019793:Fluorescein; D005456:Fluorescent Dyes / Q000032:analysis; D007091:Image Processing, Computer-Assisted; D007478:Iontophoresis; D008297:Male; D008845:Microinjections; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000648:ultrastructure*; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000032:analysis; D017966:Pyramidal Cells / Q000648:ultrastructure; D051381:Rats; D012235:Rhodamines; D013048:Specimen Handling / Q000379:methods*; D016707:Tissue Fixation