title: Rapid fixation and immunofluorescent staining of cultured cells using microwave irradiation.
abstract: Microwave irradiation during tissue fixation and immunostaining reduces the sample preparation time. Microwave irradiation also facilitates the penetration of fixatives and antibody solutions into the tissues, resulting in efficient fixation and reduction of non-specific antibody binding, respectively. Experimental procedures involving immunofluorescence microscopy are time-consuming as this method relies on antigenantibody reaction. Here, we utilized a technique involving exposure of cultured cells and tissues to intermittent microwave irradiation and immunostaining of fixed samples. Intermittent microwave irradiation during fixation reduces the required incubation time with blocking and antibody solutions, and results in good preservation of the immunoreactivity of fixed cells. Microwave irradiation also reduces the non-specific binding of fluorescein-labeled antibodies. These microwave-assisted rapid immunofluorescence techniques are useful for analysis of molecular compositions in cultured cell systems.
authors.affiliation: Department of Anatomy, Jichi Medical University, Shimotsuke-city, Tochigi, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Katoh
keywords: Cell Culture; Immunofluorescence microscopy; Immunostaining; Microwave