Microbiology 46 = Identification of Bacteria (Part-01) = Introduction of Microbial Staining Methods

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Different types of microorganisms cannot be seen by the naked eyes, so we need the help of a microscope but without the colouring matter or simply known as dyes we can not see the shape and size of bacteria, because those are colourless and transparent in nature. Live bacteria do not show the structural detail under the light microscope due to a lack of contrast. Hence staining techniques are used to produce colour contrast. Routine methods of staining of bacteria involve dying and fixing smears procedures that kill them. Bacteria have an affinity to basic dyes due to the acidic nature of their protoplasm. Staining is an Important process in microbiology. To study all structure and shape of microorganisms one must know and able to see the microorganism, and for that, it has to be coloured by utilizing simple, monochrome, negative, differential staining and other staining techniques.

Staining is a technique in which we apply a colouring agent to the sample of the microscope, so that we may observe the internal structure of the microorganism with the help of a microscope. Without applying a stain it’s very difficult to visualize the sample of the microorganism.

स्टैनिंग तकनीक ऐसी तकनीक है, जिसमे हम किसी सूक्ष्म जीवों के सैंपल को माइक्रोस्कोप की सहायता से देखने के लिए कलर का उपयोग करते हैं, क्यूंकि हम बिना कलर के सूक्ष्म जीवों को नहीं देख सकते. स्टैनिंग टेक्निक का प्रयोग कर के हम सूक्ष्म जीवों के आतंरिक संरचना को देख सकते हैं.

The structure of the organism’s cell wall determines whether the organism is gram-positive or negative. When stained with a primary stain and fixed by a mordant, some bacteria are able to retain the primary stain by resisting decolourization while others get decolourized by a decolourizer. Those bacteria which retain the primary stain are called Gram-positive and those bacteria which gets decolourized and then get counterstained are called Gram-negative.

Some staining techniques involve the application of only one dye to the sample; others require more than one dye. In simple staining, a single dye is used to emphasize particular structures in the specimen. A simple stain will generally make all of the organisms in a sample appear to be the same color, even if the sample contains more than one type of organism. In contrast, differential staining distinguishes organisms based on their interactions with multiple stains. In other words, two organisms in a differentially stained sample may appear to be different colors. Differential staining techniques commonly used in clinical settings include Gram staining, acid-fast staining, endospore staining, flagella staining, and capsule staining

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