Patent ID: 9023615
Filing Date: 2015-05-05
Classification: C12Q,G01N

Abstract:
1. A method for Gram staining bacteria on a slide using automatic staining equipment, wherein the automatic staining equipment comprises: carousel means for holding a plurality of slides, each slide having thereon a sample to be stained; means for rotating the carousel means at predetermined speeds; and means for directing and controlling application of reagents onto the slides and samples during rotation of the carousel means, and wherein the method comprises: directing a crystal violet reagent onto the slides in the carousel to contact the bacteria on the slides with the crystal violet reagent; directing water, aqueous buffer, or aqueous detergent, onto the slides in the carousel to rinse the crystal violet reagent from the slides; directing iodine reagent onto the slides in the carousel to contact the bacteria on the slides with the iodine reagent; directing water, aqueous buffer, or aqueous detergent, onto the slides in the carousel to rinse iodine reagent from the slides; directing a slow-acting decolorizing formulation and a counterstain, simultaneously or separately, onto the slides in the carousel, wherein the slow-acting decolorizing formulation comprises a slow acting decolorizing agent selected from the group consisting of 1,2-propanediol, ethylene glycol, and mixtures thereof, and wherein the slow-acting decolorizing formulation takes at least 60 seconds to remove the crystal violet reagent and the iodine reagent from a Gram negative bacteria; and directing water, aqueous buffer, or aqueous detergent, onto the slides in the carousel to rinse the slow-acting decolorizing formulation and the counterstain from the slides, wherein at least 180 seconds elapse between the step of directing a slow-acting decolorizing formulation and a counterstain, simultaneously or separately, onto the slides in the carousel and the step of directing water, aqueous buffer, or aqueous detergent, onto the slides in the carousel to rinse the slow-acting decolorizing formulation and the counterstain from the slides.