Staining Procedures The following procedure should be followed for safranin staining. To prepare the staining solution; Add 20mg safranin powder to a ml beaker. Pour 20ml distilled water in the beaker and make 0. Transfer 20mg of fast green dye in another ml beaker. Moreover, make it 0.
Filter both the staining solutions to avoid particles. To prepare slides for observation; Hydrate the slides after deparaffinization. After washing the slides, stain them with the fast green solution for minutes. Rinse the slides with 0. Stain the slides with 0. Add the primary stain crystal violet on the sample and incubate it for 1 minute.
Rinse the slide with water to remove unbound dye. Incubate the slide for 1 minute after adding the safranin stain secondary stain. Wash the slide with water for 5 seconds. Mount the slide and observe. Stain the slides in Safranin O solution for hours. Wash out the excess stain with distilled water. Counterstain with fast green staining solution for seconds. Add additional solvent to retain fast green stain as the dye evaporates with use. Rinse excess fast green solution.
Wash the slides with clearing solution by dipping for seconds. Clear the slides in xylene. Repeat the step twice. Keep the slides in the final xylene solution while you mount the coverslip. Observe the tissue sections under the microscope. Application The safranin staining is the most widely used staining technique for cell differentiation, cell-based assays, and stem cell culture. Use Of Safranin For Staining Of Frozen Sections Safranin is a cheap and safe dye which has been predominantly used to stain plant tissues for histological analysis.
Micro-Spectrophotometric Quantitation Of Glycosaminoglycan In Articular Cartilage Sections Stained With Safranin O The safranin staining has helped the researchers to develop a new micro-spectrophotometric method for the quantitation of glycosaminoglycan in cartilage tissues.
Strengths And Weaknesses The safranin stain is widely used in in-vitro diagnostics. The safranin staining is the most popular counter-stain used in medical laboratories. The safranin stain is a cheaper and safer-lab stain. It is a certified stain for chromosomes. It can be used to stain animal as well as plant cells for better cytological and histological analysis.
It offers improved accuracy in the diagnosis of frozen sections of tumor cells. It enables the researchers to perform qualitative as well as quantitative analysis of proteoglycan in cartilage matrix in the diagnosis of articular diseases. In diseases where glycosaminoglycan loss is severe, safranin is not a sensitive indicator.
Precautions Following precautionary measures should be taken while using the safranin stain. Keep the safranin powder away from heat and hot surfaces as safranin is flammable. Wear gloves and face mask while handling the safranin solution as it can cause severe eye damage and skin irritation. Store the powder in a cold or well-ventilated place.
Gram Staining Created by Monica Z. Bruckner What is Gram Staining? Show Caption. Anthrax gram stain. The Gram Stain is a differential staining technique that allows us to differentiate gram negative cells from gram positive cells based upon the chemistry and structure of their cell walls. After completing the staining protocol, gram positive cells appear purple while gram negative cells appear pink.
The gram stain utilizes crystal violet as the primary stain. This basic dye is positively charged and, therefore, adheres to the cell membranes of both gram negative and positive cells.
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