title: Improving images of fluorescent cell labeling by background signal subtraction.
abstract: The uptake and selective accumulation of fluorescent labels and drugs into organelles of cultured cells currently are widely investigated in biomedical research. In such studies, co-localization procedures are frequently used to identify the accumulation sites of compounds with biological activity. A drawback with fluorescent labeling is the autofluorescence of some cell organelles, which can hinder the precise assessment of co-localization. We report here labeling of the Golgi apparatus of A-549 cells using the photosensitizer zinc(II)-phthalocyanine (ZnPc) and co-localization with the Golgi probe NBD C6-ceramide. The blue autofluorescence signal of mitochondria can be subtracted easily from the original picture by image processing, after which the co-localization of the isolated red ZnPc signal with the green signal from the Golgi probe is considerably improved.
authors.affiliation: Department of Biology, Faculty of Sciences, Autonomous University of Madrid, Cantoblanco, 28049 Madrid, Spain. juancarlos.stockert@uam.es
authors.forename: J C
authors.initials: JC
authors.lastname: Stockert
authors.forename: A
authors.initials: A
authors.lastname: Villanueva
authors.forename: J
authors.initials: J
authors.lastname: Cristóbal
authors.forename: M
authors.initials: M
authors.lastname: Cañete
mesh_terms: D009327:4-Chloro-7-nitrobenzofurazan; D045744:Cell Line, Tumor; D002518:Ceramides; D003287:Contrast Media; D006801:Humans; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D007211:Indoles; D054833:Isoindoles; D008175:Lung Neoplasms / Q000473:pathology*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009942:Organometallic Compounds; D017319:Photosensitizing Agents; D013194:Staining and Labeling / Q000379:methods; D013382:Subtraction Technique; D017967:Zinc Compounds
chemical_list: C120266:C12-NBD-ceramide; D002518:Ceramides; D003287:Contrast Media; D007211:Indoles; D054833:Isoindoles; D009942:Organometallic Compounds; D017319:Photosensitizing Agents; D017967:Zinc Compounds; C052159:Zn(II)-phthalocyanine; D009327:4-Chloro-7-nitrobenzofurazan