6533b827fe1ef96bd1285bb2
RESEARCH PRODUCT
Labeling of Single Cells in the Central Nervous System of <em>Drosophila melanogaster</em>
Kerri-lee HarrisThomas KunzChristof RickertPaul M WhitingtonGerhard M. Technausubject
biologyGeneral Immunology and MicrobiologyGeneral Chemical EngineeringGeneral NeuroscienceCellbiology.organism_classificationCell morphologyEmbryonic stem cellMolecular biologyeye diseasesGeneral Biochemistry Genetics and Molecular BiologyCell biologyGreen fluorescent proteinlaw.inventionmedicine.anatomical_structureSingle-cell analysisConfocal microscopylawmedicinesense organsDrosophila melanogasterDevelopmental biologydescription
In this article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the lipophilic fluorescent membrane marker DiI. This method allows the visualization of neuronal cell morphology in great detail. It is possible to label any cell in the CNS: cell bodies of target neurons are visualized under DIC optics or by expression of a fluorescent genetic marker such as GFP. After labeling, the DiI can be transformed into a permanent brown stain by photoconversion to allow visualization of cell morphology with transmitted light and DIC optics. Alternatively, the DiI-labeled cells can be observed directly with confocal microscopy, enabling genetically introduced fluorescent reporter proteins to be colocalised. The technique can be used in any animal, irrespective of genotype, making it possible to analyze mutant phenotypes at single cell resolution.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2013-03-04 | Journal of Visualized Experiments |