title: Direct confocal acquisition of fluorescence from X-gal staining on thick tissue sections.
abstract: X-gal staining is a common procedure used in the histochemical monitoring of gene expression by light microscopy. However, this procedure does not permit the direct confocal acquisition of images, thus preventing the identification of labelled cells on the depth (Z) axis of tissue sections and leading sometimes to erroneous conclusions in co-localization and gene expression studies. Here we report a technique, based on X-gal fluorescence emission and mathematically-based optical correction, to obtain high quality fluorescence confocal images. This method, combined with immunofluorescence, makes it possible to unequivocally identify X-gal-labelled cells in tissue sections, emerging as a valuable tool in gene expression and cell tracing analysis.
authors.affiliation: Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: Konstantin L
authors.initials: KL
authors.lastname: Levitsky
authors.forename: Juan José
authors.initials: JJ
authors.lastname: Toledo-Aral
authors.forename: José
authors.initials: J
authors.lastname: López-Barneo
authors.forename: Javier
authors.initials: J
authors.lastname: Villadiego
mesh_terms: D000818:Animals; D005470:Fluorometry; D005697:Galactosides / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D016591:Histocytological Preparation Techniques; D007211:Indoles / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D005697:Galactosides; D007211:Indoles; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside