Combination of diOlistic labeling with retrograde tract tracing and immunohistochemistry.
Neuronal staining techniques have played a crucial role in the analysis of neuronal function. Several different staining techniques have been developed to allow morphological analyses of neurons. DiOlistic labeling, in which beads are coated with a lipophilic dye and then ballistically ejected onto brain tissue, has recently been introduced as a useful and simple means to label neurons and glia in their entirety. Although diOlistic labeling provides detailed information on the morphology of neurons, combining this approach with other staining methods is a significant advance. We have developed protocols that result in high quality diOlistically- and retrogradely-labeled or diOlistically-immunohistochemically labeled neurons. These dual-label methods require modification of fixation parameters and the restricted use of detergents for tissue permeabilization, and are readily applicable to a wide range of tracers and antibodies.
Neely
M Diana
MD
Department of Psychiatry, Vanderbilt University Medical Center, 3066 VPH, 1601 23rd Avenue South, Nashville, TN 37212, USA. diana.neely@vanderbilt.edu
Stanwood
Gregg D
GD
Deutch
Ariel Y
AY
D000818:Animals; D000831:Animals, Newborn; D019470:Biolistics / Q000379:methods*; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002232:Carbocyanines / Q000737:chemistry / Q000493:pharmacokinetics*; D048430:Cell Shape / Q000502:physiology; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry / Q000493:pharmacokinetics*; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D008845:Microinjections / Q000379:methods; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008863:Microspheres; D009434:Neural Pathways / Q000166:cytology / Q000378:metabolism; D056347:Neuroanatomical Tract-Tracing Techniques / Q000379:methods*; D009445:Neuroanatomy / Q000379:methods; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009924:Organ Culture Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods
C035005:3,3'-dihexadecylindocarbocyanine; D002232:Carbocyanines; D005456:Fluorescent Dyes