Which of the following are true for Gram-negative bacteria? Acidic and Basic Stains (Dyes): All stains (dyes) used to stain bacteria are synthetic products because they are artificially produced mainly from fractionation and recombination of coal-tar (aniline) and hence are generally called coal-tar dyes or aniline dyes. Examples: Congo Red, Nigrosin, and India Ink. 8.2K views State two ways that can confirm whether a bacterial smear has been correctly prepared or not. On the other hand, the negatively charged chromophores in acidic dyes are repelled by negatively charged cell walls, making them negative stains. 23. For example, a dye with a negative charge will be rejected by bacteria whose cell walls are also negatively charged and will therefore repel the dye. Giemsa stain is a compound dye. Acidic dyes are not very often used in Microbiology lab.except to provide background staining like Capsule staining. basic type as nucleic acid of bacterial cells attract the positive ions, e.g. Basic dyes are extensively used for staining of bacterial cells as they stain well due to abundance of ribonucleic acid (RNA) in these cells. Acidic Dyes: It is dye which has negative charge so they bind to positively charged cell structures like some proteins. Acidic dyes contains Different dyes which can be used for staining are basic/cationic dyes (methylene blue, crystal violet, malachite green, safranin) and anioninc/acidic dyes (eosin and picric acid). Dyes that are used for biological staining may be basic, neutral, or acid. Further, an example of basic dye which has amino groups as their auxochrome is Basic Blue 9 which is very popular dye and has great usage value. Basic dyes posses cations (positively charged) that combine with an acid in the stained material to form a salt . A basic dye carries a positive charge: it has either gained a hydrogen or lost a hydroxide. On the other hand, the negatively charged chromophores in acidic dyes are repelled by negatively charged cell walls, making them negative stains. Basic dyes comprise: Amino groups / Alkylamino groups (as their auxochromes) A few examples of basic dyes are: Methylene blue. The dye eosinate of methylene blue belongs to which group? stains are used commonly in microbiology to increase the contrast between microorganisms or parts of its and the background,so that it can be easily visible. Such dyes are prepared by adding the cytochrome to a chromophore. Differential stains use more than one dye. Basic dyes are usually applied to Acrylic fibres. State two ways that can confirm whether a bacterial smear has been correctly prepared or not. The best bacterial stains are aniline dyes (synthetic organic dyes made from coal-tar products). From the name, it is quite clear that it is a very simple or direct staining method that uses a single stain only. We have also developed a complete toolkit of reagents for viability PCR, a revolutionary method for determining microbial viability using qPCR. If the color portion of the dye resides in the positive ion, as in the above case, it is called a basic dye (examples: methylene blue, crystal violet, safranin). This is because opposite charges attract, basic dyes stain the negatively charged components of cells including nucleic acid & many proteins. Examples: Nigrosine, Picric acid, Eosin, Acid fuschin, India ink etc. After this type of dye ionization, the dye ion is positively charged and can combine with acidic substances to form a salt. I also have videos on the Gram Stain, Flagella Stain, Endospore Stain, Capsule Stain and more if you want to check them out! If the color portion of the dye resides in the positive ion, as in the above case, it is called a basic dye (examples: methylene blue, crystal violet, safranin). Basic dyes have their color pigment on the positive ion and therefore are attracted to the cell and stain the cell. Thus acidic conditions favor the work of acidic dye stain whereas alkaline pH works well with basic dyes. Acidic dyes are not very often used in Microbiology lab.except to provide background staining like Capsule staining. The neutral components of the cell are stained by both the dyes of wright stain and result in variable colors. Since most of the bacterial cells are negatively charged on the surface, acidic dyes can't stain them. the contrast between the cell and the background. That means it contains both acid and basic dyes in combination. That means it helps to differentiate both acidic and basic components of cells. The acid alcohol contains 3% HCl and 95% ethanol, or you can declorase with 20% H 2SO 4 8. The pH of the Cytoplasm of 0organisms. For natural dyestuffs, see Glossary of dyeing terms. This stain contains azure, methylene blue, and eosin dye. Microbiology Notes 7. Define acidic and basic dyes. 17 . Dyes or stains may be divided into 2 groups: basic and acidic. Saturate it with carbol-fuschin and heat fix by steaming over a boiling water bath or beaker for 5-10 minutes while adding more dye to the smear. Acidic dyes have their color pigment on the negative ion and therefore are repelled by the cell, but stain the background. Basic dyes consists of amino groups, or alkylamino groups, as their auxochromes. If the chromophore is the positively charged ion, the stain is classified as a basic dye; if the negative ion is the chromophore, the stain is considered an acidic dye. Staining procedure. The Gram stain is the most common differential stain used in microbiology. Here is a link to a playlist on. Acidic/Basic Dyes The most widely used histological stains differentiate between the acidic and basic components of cells and tissues. Basic dyes are extensively used for staining of bacterial cells as they stain well due to abundance of ribonucleic acid (RNA) in these cells. It is based on Ionic charges of the elements of the cell based on the concept of attraction and repulsion between the ions as well as the dyes. Acidic stains are used for background staining. Cationic dyes bids with the negatively charged cellular components such as those are present in cell membrane, whereas, anionic dye binds with positively charged . For . Principle of Giemsa stain. the simple stains: Because most cells are transparent, staining them with dyes makes them easier to see and discern. Acid dyes have negatively charged chromophores and are repelled by the bacterial surface forming a deposit aroung the organism. This means that the stain readily gives up a hydrogen ion (proton) and the chromophore of the dye becomes negatively charged. Attachment of the chromo-phore part of the dye complex to a cellular component represents the staining reaction. Basic dyes, which carry a positive charge, are more frequently supplied for staining than are negatively charged acidic dyes. Acidic dyes, such as rose bengal and eosin serve as negative stains as these are repelled by negatively charged cell walls. Eg. 1. India ink , Stains the background, not the cells Most dyes are actually basic and can be absorbed by bacteria. Phosphate groups of nucleic acids (DNA and RNA) B. stains with acidic dye eosin, has histamine and major basic protein in granules, and facilitates responses to protozoa and helminths. Cells are stained with a colored dye that makes them more visible under the light microscope. Biology Q&A Library Why are basic dyes more effective for bacterial staining than acidic dyes? Acidic Dyes carry which charge? Basic fuchsin safranin. They stain the background and leave the microbe Nigrosine and congo red are examples of acid dyes. 2. Staining . If the chromophore is the positively charged ion, the stain is classified as a basic dye; if the negative ion is the chromophore, the stain is considered an acidic dye. water - spore stain (primary stain is only slightly basic and does not stick to cells well, so simple water washing removes dye from all but the re-solidified spore coat) acid-alcohol - acid fast stain (harsh acid decolorizes even gram positives with thick peptidoglycan, leaving only the waxy-walled (mycolic acid) Mycobacterium colored. CONTENTS 1. When they are used directly on fixed bacterial smears, the contours of bacterial bodies are clearly seen. Dying is done in an acidic medium. Basic dyes are positively charged and work with negatively charged tissue components, Basic dyes are constructed from cationic salts of colored bases, and acidic salts consist of salt of a sulfuric, phenolic or carboxylic organic acid. The stain employs both acidic and basic dyes, so that the basic dye stain acidic elements of the cell, while the acidic dyes stain the fundamental components of cells. More accurately, dyes can be referred to as anionic (-) or cationic (+) and this is the convention that will be used in this manual. If the color portion is in the negatively charged ion, it is called an acidic dye (examples: nigrosin, congo red). We have also developed a complete toolkit of reagents for viability PCR, a revolutionary method for determining microbial viability using qPCR. The microorganisms are invisible to the naked eye, and to make them visible, staining is performed that gives divergence to a microscopic image. D a) upon alcohol treatment, the permeability of the cell wall increases b) crystal violet-iodine (CV-I) complex is extracted The process of adding a dye to a bacterial culture. Simple staining techniques is used to study the morphology . Acidic dyes: Nigrosin, Congo red Basic dyeshave a net positive charge and bind to components of cells and tissues that are negatively charged. Acid dyes are typically soluble in water and possess affinity for amphoteric fibers, wool, silk etc. Some are anionic dyes (negatively charged), or acidic dyes, such as nigrosin or India ink. This dye can be acidic or it can be a basic dye. Basic dyes posses cations (positively charged) that combine with an acid in the stained material to form a salt . Tilt slide 45 degrees over the sink and . Match each cell type with its description. Reagents Required. It is used to stain basic (positively charged) components such as histone protein or background. Basic dyes are positively charged and they contain cation, which when reacts with anions (-COOH -OH, nucleic acid) of the acid present in the stained cell forms a salt. If the color portion is in the negatively charged ion, it is called an acidic dye (examples: nigrosin, congo red). The charge of the dye's ions also determines if the dye is basic or acidic. Specially prepared dyes are used for such staining purposes. Basophilic staining uses basic dyes while acidophilic staining uses acidic dyes. Stains are solutions. View lab 2 microbiology.docx from BIO MISC at Bunker Hill Community College. A chromophore is also a colored compound but it does not have the properties of the dye. Acidic dye or anionic dye: In acidic dye, colored part is negatively charged and hence also called anionic dye. 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