STED Imaging in Drosophila Brain Slices.
Super-resolution microscopy is a very powerful tool to investigate fine cellular structures and molecular arrangements in biological systems. For instance, stimulated emission depletion (STED) microscopy has been successfully used in recent years to investigate the arrangement and colocalization of different protein species in cells in culture and on the surface of specimens. However, because of its extreme sensitivity to light scattering, super-resolution imaging deep inside tissues remains a challenge. Here, we describe the preparation of thin slices from the fruit fly (Drosophila melanogaster) brain, subsequent immunolabeling and imaging with STED microscopy. This protocol allowed us to image small dendritic branches from neurons located deep in the fly brain with improved resolution compared with conventional light microscopy.
Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152, Martinsried, Munich, Germany.
Sandra
S
Fendl
Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152, Martinsried, Munich, Germany.
Jesús
J
Pujol-Martí
LMU Munich, Biocenter Martinsried, Grosshadernerstr. 2, 82152, Martinsried, Munich, Germany.
Joel
J
Ryan
Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152, Martinsried, Munich, Germany.
Alexander
A
Borst
Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152, Martinsried, Munich, Germany. rkasper@neuro.mpg.de.
Robert
R
Kasper
D000818:Animals; D001921:Brain / Q000378:metabolism*; D016616:Cryoultramicrotomy / Q000379:methods; D049229:Dendritic Spines / Q000378:metabolism; D004330:Drosophila / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging; D009474:Neurons / Q000378:metabolism
Brain slice; Cryostat sectioning; Drosophila melanogaster; Immunofluorescence; STED