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Mar 14, 2024 · A mordant is a substance used to set or stabilize stains or dyes; in this case, Gram’s iodine acts like a trapping agent that complexes with the crystal violet, making the crystal violet–iodine complex clump and stay contained in thick layers of peptidoglycan in the cell walls.
- What Is Gram Staining?
- Gram Staining Objectives
- Gram Staining Principle
- Gram Staining Requirements
- Procedure of Gram Staining
- Applications of Gram Staining
- Limitations of Gram Staining
- References
Gram staining is a differential bacterial staining technique used to differentiate bacteria into Gram Positive and Gram Negative types according to their cell wall composition. It is the most widely used and the most important staining technique in bacteriology, especially in medical bacteriology. It is generally the first test performed on bacteri...
To differentiate bacteria into Gram-Positive and Gram-Negative.To study the morphological structure of bacteria.Gram staining and differentiation are based on the differences in cell wall structure and composition of bacteria. Bacteria having cell walls with a thick layer of peptidoglycan will resist decolorization of primary stain and appear violet or purple. Bacteria having a thin peptidoglycan layer with lesser cross-linkage lose primary stain during deco...
Sample bacterial colonies or suspensionGram Staining Kit (Reagents)Glass slideInoculating loopGram Staining Reagents Preparation
1. Crystal Violet Preparation 1. For preparing solution A (Crystal Violet stock solution), add 20 gm of 85% crystal violet dye in 100 ml ethanol (95%) and dissolve by mixing thoroughly. 2. For preparing solution B (Oxalate Stock solution), add 1 gm ammonium oxalate in 100 ml distilled water and dissolve by mixing thoroughly. 3. For preparing the working solution, add 1 ml of crystal violet stock solution in 10 ml distilled water and add 40 ml of oxalate stock solution. 4. Let the solution sit...
Procedure of Gram Stain Slide Preparation
1. Take a clean, clear, grease-free glass slide 2. Sterilize the inoculating loop by flaming and transfer a loop full of bacterial culture suspension in the middle of the glass slide. If culture is on a petri dish or slant, place a drop of water in the middle of the glass slide and using a sterile loop, transfer a small amount of colony and suspend with the water drop. 3. Spread the suspension with the sterile inoculating loop to prepare a thin smear. The smear must not be too thin or too thi...
Gram Staining Protocol
1. Flood crystal violet solution over fixed smear 2. After 30 – 60 seconds, pour off the CV solution and rinse with gentle running water. 3. Flood the Gram’s Iodine solution over the smear 4. Leave the iodine solution for 30 – 60 seconds and pour off the excess iodine and rinse with gentle running water 5. Shake off the excess water over the smear 6. Decolorize the smear by passing the decolorizing solution till the solution runs down in clear form. Alternatively, add a few drops of decoloriz...
Used in research to classify the bacteria into Gram-positive and Gram-negativeUsed in diagnostic labs for identification of the pathogenUsed in hospital for choosing spectrum of antibiotic for treatment before complete identification of bacteriaUsed to study the morphology of bacteriaCan’t stain Acid Fast Bacilli (Mycobacterium spp.,), and bacteria without cell wall like Mycoplasma spp.Unsuitable for minute bacteria like Ricktessia spp., Chlamydia spp.,etc.Require multiple reagents.Over-decolorization may result in the identification of false gram-negative results, whereas under-decolorization may result in the identification of false gram-positive results.Thetner, D. (2000), “Triphenylmethane and related dyes”, Kirk-Othmer Encyclopedia of Chemical Technology, Wiley, doi:10.1002/0471238961, ISBN 9780471484943.S013.pdf (himedialabs.com)Microsoft Word – IFU40168_MC_.doc (thermofisher.com)SAFRANIN: CHARACTERISTICS, USE, TECHNIQUES, TOXICITY – SCIENCE (warbletoncouncil.org)A mordant is a substance used to set or stabilize stains or dyes; in this case, Gram’s iodine acts like a trapping agent that complexes with the crystal violet, making the crystal violet–iodine complex clump and stay contained in thick layers of peptidoglycan in the cell walls.
Dec 24, 2022 · At its simplest, the actual staining process may involve immersing the sample (before or after fixation and mounting) in dye solution, followed by rinsing and observation. Many dyes, however, require the use of a mordant — a chemical compound that reacts with the stain to form an insoluble colored precipitate.
The endospore stain is used to distinguish between bacterial species that do and do not form endospores. Endospores (if present) appear bluish-green and all other cells/cell structures appear pink. The flagella stain uses the following dyes/reagents: tannic acid or potassium alum mordant, pararosaline or basic fuchsin.
Jul 16, 2012 · A mordant is a substance used to set dyes on fabrics or tissue sections by forming a coordination complex with the dye which then attaches to the fabric or tissue. [1] . It may be used for dyeing fabrics, or for intensifying stains in cell or tissue preparations. A mordant is always a polyvalent metal ion. [2] .
2 days ago · Principle: Staining is an auxiliary technique used in microscopic techniques used to enhance the clarity of the microscopic image.Stains and dyes are widely used in the scientific field to highlight the structure of the biological specimens, cells, tissues etc.
