FlyClear: A Tissue-Clearing Technique for High-Resolution Microscopy of Drosophila.
Methods Mol Biol
; 2540: 349-359, 2022.
Article
in En
| MEDLINE
| ID: mdl-35980588
ABSTRACT
Fluorescently labeled transgenic lines of Drosophila melanogaster are a powerful routine tool in fly laboratories. The possibility to fluorescently visualize individual cell populations or entire tissues and the constantly improving microscopy technologies such as two-photon or light-sheet applications, with deep tissue imaging, hold great potential to address central biological questions at an organismic level. However, strong pigmentation and the opaque nature of the D. melanogaster cuticle hinder the penetration of visible light into internal tissues, thereby limiting the application of fluorescent microscopes to analyses of the outermost surfaces of intact samples. In addition, tissue-induced light scattering and optical aberrations quickly blur the view and, hence, require tissue sectioning for further investigation. We have developed a tissue-clearing and depigmentation approach (FlyClear), which preserves endogenous fluorescent signals and is applicable to various developmental stages ranging from larvae to adult fruit flies (Pende et al. Nature communications 94731, 2018). In this chapter, we provide a detailed protocol of the experimental steps involved.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Drosophila
/
Drosophila melanogaster
Limits:
Animals
Language:
En
Journal:
Methods Mol Biol
Journal subject:
BIOLOGIA MOLECULAR
Year:
2022
Type:
Article
Affiliation country:
United States