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TY - JOUR AU - Janssens, Hilde AU - Kosman, Dave AU - Vanario-Alonso, Carlos E. AU - Jaeger, Johannes AU - Samsonova, Maria AU - Reinitz, John PY - 2005 DA - 2005/07/01 TI - A high-throughput method for quantifying gene expression data from early Drosophila embryos JO - Development Genes and Evolution SP - 374 EP - 381 VL - 215 IS - 7 AB - 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. SN - 1432-041X UR - https://doi.org/10.1007/s00427-005-0484-y DO - 10.1007/s00427-005-0484-y ID - Janssens2005 ER -