BIO TECH TEACHING KIT - Staining Techniques

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Gram staining, also called Gram's Method, is a method of differentiating bacterial species into two large groups (gram-positive and gram-negative). The name comes from its inventor, Hans Christian Gram.Gram staining consists of four components
- Primary stain (Crystal violet, methyl violet or Gentian violet)
- Mordant (Gram's Iodine)
- Decolourizer (ethyl alcohol, acetone or 11 ethanol-acetone mixture)
- Counterstain (Dilute carbol fuchsin, safranin or neutral red)

Components

• Primary Stain(RT)
• Mordant (RT)
• Decolourizer(RT)
• Secondary Stain(RT)
• Glass slides (RT)
• Filter paper (RT)
• Protocol

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Bacteria and fungi can grow across a large spectrum of environmental conditions. Even though the bacterium may grow well in the human body at 37 DegreeC at pH 7 conditions, it may also be able to withstand out-of-the-ordinary pH, temperature, and osmotic pressure. Basically, there are 3 large groups of microorganisms with respect to their temperature preference -mesophiles, thermophiles, and psychrophiles. Animal pathogens are mesophilic, growing well in the range of 20-45 DegreeC. Psychrophiles have an optimal range below mesophiles (even below freezing), while thermophiles grow at temperatures over 45 DegreeC.


Components

• Bacterial Culture (4 DegreeC)
• LB Broth (RT)
• Agar(RT)
• Protocol

MOQ 5 Exps

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Endospore staining is a technique used in bacteriology to identify the presence of endospores in a bacterial sample, which can be useful for classifying bacteria. Within bacteria, endospores are quite protective structures used to survive extreme conditions, but this protective nature makes them difficult to stain using normal techniques. Special techniques for endospore staining include the Schaeffer–Fulton stain and the Moeller stain. A good stain to use for spore staining is malachite green. It takes a long time for the spores to stain due to their density, so time acts as the mordant when doing this differential stain; the slide with the bacterium should be soaked in malachite green for at least 30 minutes. Water acts as the decolorizer. A counterstain to differentiate the vegetative cells is 0.5% safranin. Types of endospores that could be identified are free endospores, central endospores, central and swollen endospores, and subterminal endospores.


Components

• Primary stain (RT)
• Counter stain (RT)
• Glass slide (RT)
• Bacillus culture (4°C)
• Filter paper (RT)
• Protocol

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The use of single stain to colour the bacteria is commonly called as monochrome staining. (Mono-single, Chrome-color)The method of staining utilizes the knowledge that bacteria possess slight negative charge. The negative charged group of bacterial cell surface produces attraction between basic stains. The stains used in this method include methylene blue, crystal violet,etc .


Components

• Bacterial strain (4 DegreeC)
• Methylene blue stain (RT)
• Glass slide (RT)
• Absorbent paper (RT)
• Protocol