Fluorescent visualization of macromolecules in Drosophila whole mounts.
The ability to determine the expression dynamics of individual genes "in situ" by visualizing the precise spatial and temporal distribution of their products in whole mounts by histochemical and immunocytochemical reactions has revolutionized our understanding of cellular processes. Drosophila developmental genetics was one of the fields that benefited most from these technologies, and a variety of fluorescent methods were specifically designed for investigating the localization of developmentally important proteins and cell markers during embryonic and post embryonic stages of this model organism. In this chapter we present detailed protocols for fluorescence immunocytochemistry of whole mount embryos, imaginal discs, pupal retinas, and salivary glands of Drosophila melanogaster, as well as methods for fluorescent visualization of specific subcellular structures in these tissues.
Ramos
Ricardo Guelerman Pinheiro
RG
Department of Cell and Molecular Biology, Faculdade de Medicina de Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
Machado
Luciana Claudia Herculano
LC
Moda
Livia Maria Rosatto
LM
D000818:Animals; D004331:Drosophila melanogaster / Q000033:anatomy & histology / Q000737:chemistry* / Q000166:cytology / Q000196:embryology*; D004625:Embryo, Nonmammalian / Q000737:chemistry*; D005123:Eye / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis; D007150:Immunohistochemistry / Q000379:methods*; D007211:Indoles / Q000032:analysis; D007814:Larva / Q000033:anatomy & histology; D008856:Microscopy, Fluorescence / Q000379:methods*; D010590:Phalloidine / Q000032:analysis; D011679:Pupa / Q000033:anatomy & histology; D012160:Retina / Q000737:chemistry; D012469:Salivary Glands / Q000737:chemistry; D014404:Tubulin / Q000032:analysis
D005456:Fluorescent Dyes; D007211:Indoles; D014404:Tubulin; D010590:Phalloidine; C007293:DAPI