PUBLICATION

Cellular resolution expression profiling using confocal detection of NBT/BCIP precipitate by reflection microscopy

Authors
Jékely, G., and Arendt, D.
ID
ZDB-PUB-070711-35
Date
2007
Source
Biotechniques   42(6): 751-755 (Journal)
Registered Authors
Arendt, Detlev
Keywords
none
MeSH Terms
  • Alkaline Phosphatase/metabolism
  • Animals
  • Annelida/cytology
  • Annelida/embryology
  • Chemical Precipitation
  • Drosophila/embryology
  • Embryo, Nonmammalian
  • Immunohistochemistry
  • In Situ Hybridization
  • Indicators and Reagents/chemistry*
  • Indoles/chemistry*
  • Microscopy, Confocal*
  • Nitroblue Tetrazolium/chemistry*
  • Sensitivity and Specificity
  • Zebrafish/embryology
PubMed
17612299 Full text @ Biotechniques
Abstract
The determination of gene expression patterns in three dimensions with cellular resolution is an important goal in developmental biology. However the most sensitive, efficient, and widely used staining technique for whole-mount in situ hybridization (WMISH), nitroblue tetrazolium (NBT)/5-bromo-4-chloro-3-indolyl phosphate (BCIP) precipitation by alkaline phosphatase, could not yet be combined with the most precise, high-resolution detection technique, confocal laser-scanning microscopy (CLSM). Here we report the efficient visualization of the NBT/BCIP precipitate using confocal reflection microscopy for WMISH samples of Drosophila, zebrafish, and the marine annelid worm, Platynereis dumerilii. In our simple WMISH protocol for reflection CLSM, NBT/BCIP staining can be combined with fluorescent WMISH, immunostainings, or transgenic green fluorescent protein (GFP) marker lines, allowing double labeling of cell types or of embryological structures of interest. Whole-mount reflection CLSM will thus greatly facilitate large-scale cellular resolution expression profiling in vertebrate and invertebrate model organisms.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping