Alternate gram staining technique using a fluorescent lectin. Blood is composed of the blood cells which accounts for 45% of the blood tissue by volume, with the remaining 55% of the volume composed of plasma, the liquid. Microbiology module common staining technique microbiology 24 notes indeterminate groups as well. The application of molecular technology in medicine is almost endless, some of the applications of. Biochemical tests the sole purpose of biochemical test in microbiology is to differentiate between various classes of bacteria by studying their enzymatic activities. To perform this staining, it requires the use of a single stain only. Heatfixed bacterial smears refer staining techniques in microbiology 1st edition by dr. Make a thin film of the material on a clean glass slide, using a. Staining simply means coloring of the micro organisms with the dye that emphasizes and elucidate different important structures of microorganisms including bacteria, virus, protozoa and etc. Gram staining is the common, important, and most used differential staining techniques in microbiology, which was introduced by danish bacteriologist hans christian gram in 1884. Basic techniques in biochemistry, microbiology and molecular biology pp 111116 cite as. Cover different classification schemes for grouping bacteria, especially the use of the gram stain 2.
Methods of culturing microorganisms 3 five basic techniques 4 fig. A detailed discussion of the different types of physical and chemical methods of sterilization is provided in chapter 19 physical and chemical methods of control. Microbiology module common staining technique microbiology 22 notes 2. Other differential staining methods include the endospore stain to identify endosporeforming bacteria, the acidfast stain to discriminate mycobacterium species. The most commonly used instrument in the microbiology laboratory for the sterilization of media and glassware is the autoclave. Types of staining techniques used in microbiology and their. Gram stain protocols american society of microbiology. Objectives to learn the technique of smear preparation. In microbiology the concept of staining is very important because it highlights the structures of microorganisms allowing them to be seen under a microscope. Different types of microscopes cell is the basic unit of life. It is a technique that is widely used for the examination of cells, tissues and cellular components.
Basic microbiologymlsbmic224 staining methods l 16, 17. This test differentiate the bacteria into gram positive and gram negative bacteria, which helps in the classification and differentiations of microorganisms. In microbiology, differential staining techniques are used more often than simple stains as a means of gathering information about bacteria. Wikimedia commons has media related to microbiology techniques. Types of different staining techniques of microorganisms. It was developed by danish microbiologist hans christian gram in 1884 as an effective method to distinguish between bacteria with different types of cell walls, and even today it remains one of the most frequently used staining techniques. Gram staining is a differential staining technique that differentiates bacteria into two groups. This staining technique is used to demonstrate the presence of acid fast bacilli. Virology techniques introduction virology is a field within microbiology that encompasses the study of viruses and the diseases they cause.
This staining technique divides bacteria into two groups namely acidfast and non acidfast. Staining tray, wash bottles, inoculation loop microscopic glass. Gram staining procedure animation microbiology principle, procedure, interpretation duration. Histologic staining options available include, but are not limited to. General microbiology morphology of bacterial cell, staining procedures for bacteria, culture media for bacteria, cultivation and isolation techniques for bacteria, preparation of pure cultures.
Invivo staining is the process of dyeing living tissue in vivo means in life as contrasted to invitro staining. Cell staining in microscopy types, techniques, preparations. Then apart from all this some other staining techniques are there for identification of fungus like lactophenol cotton blue staining, and many other techniques. Stains and dyes are frequently used in histology the study of tissue under the microscope and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses disease at a microscopic level. Molecular techniques in clinical microbiology molecular biology is the science of biomolecules.
Staining is a supplementary method that gives divergence to the microscopic image for better vision under the microscope. This technique was introduced in 1884 by danish physician christian gram. The most widely used staining procedure in microbiology is the gram stain, discovered by the danish scientist and physician hans christian joachim gram in 1884. Another differential stain technique is the acidfast technique. Mycobacterium species resist the effect of the acid. Staining microscopic specimens microbiology lumen learning. Blood cells are the cells which are produced during hematopoiesis and found mainly in the blood. Several different types of staining procedure are known today which. The following 61 pages are in this category, out of 61 total. This technique differentiates species of mycobacterium from other bacteria. Module morphology and classification of bacteria microbiology 10 notes stains. Microscopy refers to the practice that involves the use of a microscope for the purposes of observing small scale structures that cannot be viewed using the naked eye and often cell staining is necessary as s tructures are difficult to discern due to insufficient contrast. The extremely minute size of this wonderful structure remained an obstacle for. Different types of staining methods are used to make the cells and their internal.
Microscopy using the microscope 22 stained preparations 23 making a smear 23 a simple stain 24 a differential stain. To learn the techniques of gram staining, nigrosin staining and koh test. Composition, types and mechanism of staining composition. The word gram is always spelled with a capital, referring to hans christian gram, the inventor of gram staining gram staining principles gram staining is used to determine gram status to classify bacteria broadly. Types of staining techniques used in microbiology and. The purpose of this lesson is to provide a general overview of laboratory techniques used in the. Stains may be used to define and examine different types of microbes, various stages of cellular life e. This list may not reflect recent changes learn more.
Instruments used in microbiology lab with principle and uses. Uk standards for microbiology investigations issued by the standards unit, public health england. Bacteria have nearly the same refractive index as water, therefore, when they. Microscopic slides endospores a stained preparation of bacillus subtilis showing endospores as uncoloured and the vegetative cell as. Methylene blue, polychrome blue or carbol fuchsin to act as simple stain. Testing sensitivity to antimicrobial substances 21 2. Hematoxylin has a deep bluepurple color and stains nucleic acids by a complex, incompletely understood reaction. Differential staining methods, which typically require more than one stain and several steps, are referred to as such because they permit the differentiation of cell types or cell structures. Already available was a staining method designed by robert. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. This manual has been written for a righthanded person.
These are of two types, namely direct and indirect staining. Fluorescencelabeled wheat germ agglutinin binds specifically to nacetylglucosamine in the outer peptidoglycan layer of grampositive bacteria. Different types of staining methods are used to make the cells and. Gram staining helps to identify bacterial pathogens in specimens and cultures by their gram reaction gram positive and gram negative and morphology coccirod. Negatively charged acid radicals imparts color in eosin, acid fuchsine, malachite green, nigrosin. Cell staining in microscopy types, techniques, preparations and procedures. Heat or a lipid solvent is used to carry the first stain, carbolfuchsin, into the cells. Basic microbiology notes online microbiology notes. The instruments used in the microbiology labs include a bunch of different kinds of instruments required for a lot of different processes conducted within those laboratories. Even though the term biomolecules includes all molecules such as proteins, fatty acids etc, it is refers to nucleic acid these days. Different types of staining methods are used to make the cells and their internal structures more visible under the light microscope. Bhanu prakash usmle, fmge and neet pg 14,870 views. Staining tray, wash bottles, inoculation loop microscopic glass slides, bunsen burner, blotting papers, microscopes. Stains may be used to define biological tissues highlighting.
Gram staining primary stain step 1 crystal violet is the primary stain used in the gram staining process it is not specific for gram positive cells and it stains everything the same color purple. Chemically the cell wall is composed of peptidoglycan. Some staining techniques, such as the gram and ziel neelsen stains, although of great diagnostic value because of their differential staining properties for specific bacteria, reveal little. There are two types pili namely nonsex pili common pili eg. Gram stain the previous lab introduced simple staining techniques that enable microbiologists to observe the morphological characteristics of bacteria. Hematoxylin and eosin staining technique functions to recognize different types of tissues and their morphological changes, especially in cancer diagnosis. Then the cells are washed with a dilute acidalcohol solution. Mycobacterium and many nocardia species are called acidfast because. It determines the cell shape, size and arrangement of the microorganism. There are a variety of staining methods like simple, differential and special staining that are used in labs to examine the bacteria. The most common type of acid fast bacteria is mycobacteria. It is one of the most important and widely used differential staining techniques in microbiology.
In the laboratory, viruses have served as useful tools to better understand cellular mechanisms. The steps of the gram stain procedure are listed below and illustrated in table 1. Gram stain is a very important differential staining techniques used in the initial characterization and classification of bacteria in microbiology. Introduction as bacteria consist of clear protoplasmic matter, differing but slightly in refractive index from the medium in which they are growing, it is difficult with the ordinary microscope, except when special methods of illumination are used, to set them in the unstained condition.
S ome staining techniques, such as the gram and ziel neelsen stains, although of great diagnostic value because of their differential staining properties for specific bacteria, reveal li ttle. Staining techniques in microbiology linkedin slideshare. Introduction to microbiology culture techniques youtube. Stains are salts composed of a positive ion cation and a negative ion anion. In microbiology the concept of staining is very important because it highlights the structures of microorganisms allowing them to be seen under a microscope simple and. Sep 19, 2016 gram staining procedure animation microbiology principle, procedure, interpretation duration. Bacterial classification, structure and function introduction the purpose of this lecture is to introduce you to terminology used in microbiology. Stain or dye is the synthetic chemical which is derived from nitrobenzene or aniline. The acidfast microorganisms retain the pinkred color because carbofuschsin is more soluble in the cell wall lipids than in acidalcohol.
Microbiological laboratory techniques clinical gate. Staining techniques in some cases, staining is unnecessary, for example when microorganisms are very large or when motility is to be studied, and a drop of the microorganisms can be placed directly on the slide and observed. Coloring microorganisms with a dye that emphasizes certain structures. Some staining techniques, such as the gram and ziel neelsen stains, although. The previous lab introduced simple staining techniques that enable. To use and relate the gram stain to the study of bacterial cell morphology, and as an. Sep 25, 2019 stains may be used to define and examine different types of microbes, various stages of cellular life e.
Staining techniques in microbiology a presentation by g. Staining of bacterial cells objective to learn the techniques of smear preparation, gram staining, nigrosin staining and correlating the results of gram staining with koh test. Some staining techniques involve the application of only one dye to the. The peptidoglycan layer of gramnegative bacteria is covered by a membrane and is not labeled by the lectin. Make a thin film of the material on a clean glass slide, using a sterile loop or swab for viscous specimens. Feb 03, 2015 staining techniques in microbiology 1. Introduction observation of bacteria with conventional bright field microscopy yields relatively little useful information. Staining methods simple staining, negative staining, grams. Medical laboratory sciences medical bacteriology mls. Acknowledgments uk standards for microbiology investigations uk smis are developed under the auspices of public health england phe working in partnership with the national. Staining, therefore, is of primary importance for the.
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