title: An investigation of optimal gold particle size for immunohistological immunogold and immunogold-silver staining to be viewed by polarized incident light (EPI polarization) microscopy.
abstract: We investigated the optimal gold particle size for use with polarized incident light (epi polarization) microscopy with immunogold immunohistological preparation in both immunogold indirect (IGS) and silver-enhanced immunogold-silver staining (IGSS) techniques. A range of gold particle sizes from 5 nm-40 nm was used along with tissue of known immunoreactivity with a well-characterized primary monoclonal antibody. Checkerboard titrations were carried out for each technique and for each particle size. The preparations were viewed using a standard polarized incident light microscope and assessed in a semi-quantitative manner. Adequate visualization of gold particles was achieved using the indirect staining method only with a particle size of 40 nm. With silver enhancement (IGSS), particles of all sizes were clearly seen. However, 5-nm particles were considered optimal for this method because of reduced background staining, high titration of antisera possible, and crisp localization of the visual signal.
authors.affiliation: Department of Histopathology, Queen's Medical Centre, University Hospital, Nottingham, United Kingdom.
authors.forename: I O
authors.initials: IO
authors.lastname: Ellis
authors.forename: J
authors.initials: J
authors.lastname: Bell
authors.forename: J D
authors.initials: JD
authors.lastname: Bancroft
mesh_terms: D006046:Gold; D006801:Humans; D007150:Immunohistochemistry; D008859:Microscopy, Polarization; D010316:Particle Size; D012834:Silver; D013270:Stomach / Q000033:anatomy & histology
chemical_list: D012834:Silver; D006046:Gold