Detection and Quantification of the Bicoid Concentration Gradient in Drosophila Embryos

Feng He; Honggang Wu; David Cheung; Jun Ma

doi:10.1007/978-1-4939-8772-6_2

Detection and Quantification of the Bicoid Concentration Gradient in Drosophila Embryos

Methods Mol Biol. 2018:1863:19-27. doi: 10.1007/978-1-4939-8772-6_2.

Authors

Feng He^{1

2

3}, Honggang Wu^{4

5}, David Cheung^{6

7}, Jun Ma^{8

9

10

11

12}

Affiliations

¹ Division of Medical Genetics and Genomics, Children's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
² Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
³ Key Laboratory of Pollinating Insect Biology of the Ministry of Agriculture, Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing, China.
⁴ State Key Laboratory of Brain and Cognitive Sciences, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
⁵ University of Chinese Academy of Sciences, Beijing, China.
⁶ Division of Developmental Biology, Cincinnati Children's Research Foundation, Cincinnati, OH, USA.
⁷ Division of Biomedical Informatics, Cincinnati Children's Research Foundation, Cincinnati, OH, USA.
⁸ Division of Medical Genetics and Genomics, Children's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China. jun_ma@zju.edu.cn.
⁹ Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China. jun_ma@zju.edu.cn.
¹⁰ Division of Developmental Biology, Cincinnati Children's Research Foundation, Cincinnati, OH, USA. jun_ma@zju.edu.cn.
¹¹ Division of Biomedical Informatics, Cincinnati Children's Research Foundation, Cincinnati, OH, USA. jun_ma@zju.edu.cn.
¹² Laboratory of Systems Developmental Biology, Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China. jun_ma@zju.edu.cn.

PMID: 30324590
DOI: 10.1007/978-1-4939-8772-6_2

Abstract

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.

Keywords: Bicoid; Drosophila; Fluorescent immunostaining; Morphogen gradient.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Drosophila Proteins / metabolism*
Drosophila melanogaster / embryology*
Drosophila melanogaster / metabolism*
Embryo, Nonmammalian / cytology
Embryo, Nonmammalian / metabolism*
Female
Homeodomain Proteins / metabolism*
Image Processing, Computer-Assisted / methods*
Morphogenesis*
Spectrometry, Fluorescence
Trans-Activators / metabolism*

Substances

Drosophila Proteins
Homeodomain Proteins
Trans-Activators
bcd protein, Drosophila