Detection and Quantification of the Bicoid Concentration Gradient in Drosophila Embryos
We describe methods for detecting and quantifying the concentration gradient of the morphogenetic protein Bicoid through fluorescent immunostaining in fixed Drosophila embryos. We introduce image-processing steps using MATLAB functions, and discuss how the measured signal intensities can be analyzed to extract quantitative information. The described procedures permit robust detection of the endogenous Bicoid concentration gradient at a cellular resolution.
Key wordsFluorescent immunostaining Morphogen gradient Bicoid Drosophila
The protocol described in this chapter was developed at the Cincinnati Children’s Hospital Research Foundation. We acknowledge the invaluable contributions of Ying Wen in developing the protocol. FH acknowledges support of Institute of Apicultural Research, Chinese Academy of Agricultural Sciences. HW acknowledges the generous support of his current supervisor Dr. Renjie Jiao. JM is a National “Thousand Talents” Scholar of China and acknowledges support of Zhejiang University. Feng He and Honggang Wu contributed equally to this work.
- 17.Cheung D, Miles C, Kreitman M, Ma J (2014) Adaptation of the length scale and amplitude of the Bicoid gradient profile to achieve robust patterning in abnormally large Drosophila melanogaster embryos. Development 141(1):124–135. https://doi.org/10.1242/dev.098640CrossRefPubMedPubMedCentralGoogle Scholar