TY - JOUR
T1 - Diolistic labeling in fixed brain slices
T2 - Phenotype, morphology, and dendritic spines
AU - Staffend, Nancy A.
AU - Meisel, Robert L.
PY - 2011
Y1 - 2011
N2 - Identifying neuronal morphology is a key component in understanding neuronal function. Several techniques have been developed to address this issue, including Golgi staining, electroporation of fluorescent dyes, and transfection of fluorescent constructs. Ballistic delivery of transgenic constructs has been a successful means of rapidly transfecting a nonbiased population of cells within tissue or culture. Recently, this technique was modified for the ballistic delivery of dye-coated gold or tungsten particles, enabling a nonbiased, rapid fluorescent membrane labeling of individual neurons in both fixed and nonfixed tissue. This unit outlines a step-by-step protocol for the ballistic method of dye delivery ("DiOlistic" labeling) to fixed tissue, including optimal tissue fixation conditions. In addition, a protocol for coupling "DiOlistic" labeling with other immunofluorescent methods is detailed, enabling the association of neuronal morphology with a specific cellular phenotype.
AB - Identifying neuronal morphology is a key component in understanding neuronal function. Several techniques have been developed to address this issue, including Golgi staining, electroporation of fluorescent dyes, and transfection of fluorescent constructs. Ballistic delivery of transgenic constructs has been a successful means of rapidly transfecting a nonbiased population of cells within tissue or culture. Recently, this technique was modified for the ballistic delivery of dye-coated gold or tungsten particles, enabling a nonbiased, rapid fluorescent membrane labeling of individual neurons in both fixed and nonfixed tissue. This unit outlines a step-by-step protocol for the ballistic method of dye delivery ("DiOlistic" labeling) to fixed tissue, including optimal tissue fixation conditions. In addition, a protocol for coupling "DiOlistic" labeling with other immunofluorescent methods is detailed, enabling the association of neuronal morphology with a specific cellular phenotype.
KW - Dendritic spine
KW - Diolistic
KW - Gene gun
KW - Neuronal morphology
UR - http://www.scopus.com/inward/record.url?scp=79959551373&partnerID=8YFLogxK
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U2 - 10.1002/0471142301.ns0213s55
DO - 10.1002/0471142301.ns0213s55
M3 - Article
C2 - 21462159
AN - SCOPUS:79959551373
SN - 1934-8584
VL - Chapter 2
JO - Current protocols in neuroscience / editorial board, Jacqueline N. Crawley ... [et al.]
JF - Current protocols in neuroscience / editorial board, Jacqueline N. Crawley ... [et al.]
IS - SUPPL.55
M1 - 2.13
ER -