Permeabilization, Staining and Culture of Living Drosophila Embryos
作者:
StreckerTeresa R.,
McGheeSean,
ShihShane,
HamDebby,
期刊:
Biotechnic&Histochemistry
(Taylor Available online 1994)
卷期:
Volume 69,
issue 1
页码: 25-30
ISSN:1052-0295
年代: 1994
DOI:10.3109/10520299409106257
出版商: Taylor&Francis
关键词: Drosophilaembryo;permeabilization;octane;hexane;culturein vivo
数据来源: Taylor
摘要:
The organic solvent octane has been used routinely to permeabilize the hydrophobic vitelline membrane surrounding theDrosophilaembryo, thereby allowing the movement of small molecules into the egg. We present evidence that hexane is a more effective permeabilizing agent than octane and compare the effects of these solvents on uniformity of permeabilization and embryonic viability. The ability of each solvent to make the embryo accessible to a range of biological stains was compared. The effect of octane versus hexane permeabilization on subsequent embryonic viability was measured at seven different stages during early embryogenesis. We found that although hexane is a superior solvent for permeabilizing the vitelline membrane, it decreases the viability of embryos exposed between 0 and 3 hr of age. Older embryos treated with either hexane or octane are usually viable. We also showed that molecules with a molecular mass of 984 Daltons or more did not diffuse into the embryo following treatment with either hexane or octane. Results presented here challenge a phase-partition model that has been proposed previously to explain the molecular basis of permeabilization of theDrosophilaegg. An alternative model is described as well as an optimized protocol for permeabilizing and stainingDrosophilaembryos at any stage during early embryogenesis while maintaining viability for subsequent culture.
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