Observations & Results On Negative Staining Mix well the Specimen with the Dye using the sterilized straight wire and spread evenly over an area of about 1 – 2 cm.Īllow the smear to Air dry and then observe under the microscope at High power objective (45X) and oil immersion (100X) objectives. Now, with the help of Sterilized inoculating loop transfer a small portion of the specimen to the slide containing a Drop of Dye. Take a drop of nigrosine or India ink and place it in the middle of the Clean & Grease free Glass slide. Then move the spreader slide smoothly and rapidly forward over the specimen slide, drawing the dye mixture behind it into a thin film. Move the slide toward the drop of the specimen-dye mixture until the contact is made with the drop at the specific angle. Now, take another Microscopic Glass slide, place it near to the specimen-dye mixture at an angle of about 30° – 45°. Mix well the culture with dye taken on the Glass slide. Take a small portion of the bacterial colony with the help of sterilized Inoculating loop. Take a clean, Dry, Scratch and Grease free Microscopic glass slide and place a drop of India ink or Nigrosin on it at one end near the edge. The Procedure Of Negative Staining Technique Here is the list of Materials Required For The Negative Staining Method Hence in Negative staining Background is stained instead of Bacterial cells. Instead, these stains either produce a deposit around the bacteria or produce a dark background so that the bacteria appear as the unstained cell with a clear area around them. The above-mentioned stains do not penetrate and stains the bacterial cell due to repulsion between the negative charge of the stain and the negatively charged bacterial cell wall. In Negative staining technique, an acidic stain such as Nigrosin, India ink, Eosin or Congo red is used in which the bacterial culture or the specimen is mixed well and then spread over the Microscopic glass slide to form a thin smear. What is the Principle Of Negative Staining Technique? Moreover, Negative staining is the only staining technique in which bacterial cells are not stained but they are made visible against a dark background as colorless bodies. Negative staining is also good for viewing capsules but preferably Capsule staining is done in that cases.īriefly, in negative staining technique, the background is stained and the bacterial cell remains colorless. some of the spirochetes) or when it is desirable to confirm observations made on the shape and size or Morphology of the bacteria observed either in a wet mount or hanging drop preparations etc. The negative staining is preferred in cases when the bacterium does not stain well (e.g. Sometimes it is more convenient to determine the overall bacterial cell morphology without any use of harsh staining or heat fixing techniques that changes the shape of the bacterial cells. Thanks What is Negative Staining Technique? If you have any queries, don’t forget to mention in Comments…. Hey, Good to see you here □ …… In this Article, We’re gonna discuss the Negative Staining Technique….
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