A high-throughput method for quantifying gene expression data from early Drosophila embryos
A high-throughput method for quantifying gene expression data from early Drosophila embryos
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DOI：
10.1007/s00427-005-0484-y
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发表时间：
2005-07-01
影响因子：
2.4
通讯作者：
Reinitz, J
中科院分区：
文献类型：
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作者：
Janssens, H;Kosman, D;Reinitz, J
我们描述了一种自动化高通量方法，通过免疫荧光测量果蝇胚盘胚胎中单个核的蛋白质水平。该方法由组装成图像处理管道的一系列特定算法组成。该管道将用荧光标记抗体染色的胚胎的共聚焦扫描转换为文本文件。该文本文件包含每个核的数字标识符、其质心坐标以及该核中三种蛋白质的平均浓度。该方法的核心算法组件是通过边缘检测自动识别核，并使用分水岭作为纠错步骤。该方法提供细胞分辨率的高通量定量。
We describe an automated high-throughput method to measure protein levels in single nuclei in blastoderm embryos of Drosophila melanogaster by means of immunofluorescence. The method consists of a chain of specific algorithms assembled into an image processing pipeline. This pipeline transforms a confocal scan of an embryo stained with fluorescently tagged antibodies into a text file. This text file contains a numerical identifier for each nucleus, the coordinates of its centroid, and the average concentrations of three proteins in that nucleus. The central algorithmic component of the method is the automatic identification of nuclei by edge detection with the use of watersheds as an error-correction step. This method provides high-throughput quantification at cellular resolution.