Preparation of slides and coverslips for microscopy

INTRODUCTIONIt is imperative that the slides and coverslips used in fluorescence microscopy procedures be extremely clean. Although coverslips look clean, especially when a new box is first opened, they may have a thin film of grease on them that will not allow tissue culture cells to adhere well and that may interfere with some processing steps in certain protocols. Therefore, coverslips should routinely be washed with acid or base solutions to rid them of this film. Commercial precleaned slides are also likely to be dirty and must be washed prior to use. This protocol describes various approaches for cleaning slides and coverslips and sterilizing them for cell culture, as well as methods for subbing slides. In the subbing procedure, slides are coated with gelatin, aminoalkylsilane, or poly-L-lysine solution to promote the adhesion of cells or tissues to the glass surface. Gelatin or aminoalkylsilane is usually used for tissue sections or small organisms, whereas poly-L-lysine is routinely used for cultured cells.