Uk standards for microbiology investigations issued by the standards unit, public health england. Microbiology module common staining technique microbiology 22 notes 2. Choose from 500 different sets of staining techniques microbiology flashcards on quizlet. Scope of document the document covers the methods for the staining procedures commonly used in clinical microbiology laboratories for the identification of pathogens and dyes stains. It is one of the most important and widely used differential staining techniques in microbiology. By exploiting this phenomenon, an alternative gram staining technique has been developed.
Biochemical tests the sole purpose of biochemical test in microbiology is to differentiate between various classes of bacteria by studying their enzymatic activities. Staining techniques in microbiology linkedin slideshare. Gram staining differentiates the bacteria into 2 groups. To study biological process like phagocytosis, molecular transport across plasma membrane. It determines the cell shape, size and arrangement of the microorganism. Gram stain report 15 pts assignment each student will have the opportunity to submit one example of a gram stain with a brief written report describing the staining procedure and the stain itself. Stains and dyes are frequently used in biological tissues for viewing, often with the aid of different microscopes.
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. 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. Choose from 500 different sets of microbiology staining techniques flashcards on quizlet. This test differentiate the bacteria into gram positive and gram negative bacteria, which helps in the classification and differentiations of microorganisms. Procedure and uses august 1, 2014 acharya tankeshwar 0 bacteria are easier to detect in broth culture if the preparation is stained using toluidine blue 0. There are many different stains and staining procedures used in microbiology. A good report will likely necessitate a minimum of 2 pages doublespaced and is limited to a 3page maximum. Included in the staining procedure descriptions are explanations of the organisms that the stains identify. Neelsen in 1883 used ziehls carbolfuchsin and heat then decolorized with. This manual has been written for a righthanded person. Alternate gram staining technique using a fluorescent lectin. Differential staining techniques viewing bacterial cells. To learn the techniques of gram staining, nigrosin staining and koh test. There are a variety of staining methods like simple, differential and special staining that are used in labs to examine the bacteria.
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. Gram staining is a differential staining technique that differentiates bacteria into two groups. Staining can be performed with basic dyes such as crystal violet or methylene blue, positively charged dyes that are attracted to the negatively charged materials of the microbial cytoplasm. Hence, clinicians should be acquainted with the techniques of taking specimens, and understand the. Microbiology module common staining technique microbiology 20 notes 2 common staining technique 2. So this method is also called ziehlneelsen staining techniques. Jan 04, 2010 included in the staining procedure descriptions are explanations of the organisms that the stains identify.
This technique was introduced in 1884 by danish physician christian gram. Some staining techniques involve the application of only one dye to the sample. Microscopy using the microscope 22 stained preparations 23 making a smear 23 a simple stain 24 a differential stain. This manual, which explains the basic techniques necessary to carry out microbiology experiments safely and effectively, is intended as a guide for teachers and technicians. 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. The most commonly used instrument in the microbiology laboratory for the sterilization of media and glassware is the autoclave. Introduction observation of bacteria with conventional bright field microscopy yields relatively little useful information. Make a thin film of the material on a clean glass slide, using a. An optimized staining technique for the detection of gram. 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. Reagents for histological techniques, haematology and. Microbiological laboratory techniques clinical gate.
Hence, clinicians should be acquainted with the techniques of taking specimens, and understand the principles and techniques. Hematoxylin and eosin staining technique functions to recognize different types of tissues and their morphological changes, especially in cancer diagnosis. A stain is a dye consisting of a colored ion a chromophore and a counter ion to balance the charge. Attachment of the chromo phore part of the dye complex to a cellular component represents the staining reaction. Gram stain is a very important differential staining technique used in the initial characterization and classification of bacteria in microbiology. Objectives to learn the technique of smear preparation. Staining microscopic specimens microbiology lumen learning. Microbiology module common staining technique microbiology 24 notes indeterminate groups as well. Learn microbiology staining techniques with free interactive flashcards. Diagnostic microbiology this section contains 15 randomly generated multiple choice questions which will check your understanding regarding various types of microscopes bright field, dark field, phase contrast, fluorescence, scanning electron, transmission electron etc and staining techniques e.
Coloring microorganisms with a dye that emphasizes certain structures. These are of two types, namely direct and indirect staining. Diagnostic microbiology and laboratory methods pocket. Gram staining gram staining is most widely used differential staining in microbiology. Introduction to microbiology culture techniques youtube.
Testing sensitivity to antimicrobial substances 21 2. 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. This staining technique is used to demonstrate the presence of acid fast bacilli. It is a technique that is widely used for the examination of cells, tissues and cellular components. Module context this module introduces microbiological laboratory techniques to be used for analysis of coliforms bacteria can be used as indicators of pollution.
To use and relate the gram stain to the study of bacterial cell morphology, and as an. In simple staining, a single dye is used to emphasize particular structures in the specimen. Gram staining helps to identify bacterial pathogens in specimens and cultures by their gram reaction gram positive and gram negative and morphology coccirod. 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. Stains are salts composed of a positive ion cation and a negative ion anion. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. The peptidoglycan layer of gramnegative bacteria is covered by a membrane and is not labeled by the lectin. 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. Diagnostic microbiology involves the study of specimens taken from patients suspected of having infections.
Accurate gram staining of american type culture collection strains of these anaerobic bacteria is possible by implementing fixing and staining techniques within a gloveless anaerobic chamber. The microbiology society, in association with the microbiology in schools advisory committee misac. Cell staining in microscopy types, techniques, preparations and procedures. Special stains differential stains basic techniques of. To perform this staining, it requires the use of a single stain only. Fluorescencelabeled wheat germ agglutinin binds specifically to nacetylglucosamine in the outer peptidoglycan layer of grampositive bacteria. Comparison of two silver staining techniques for detecting. The microscope is a very important tool in microbiology, but there are limitations when it comes to using one to observe cells in general and bacterial cells in particular. 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. Staining procedures that use one dye to increase contrast between specimen and. Stains may be used to define biological tissues highlighting. Methods of culturing microorganisms 3 five basic techniques 4 fig. Bhanu prakash usmle, fmge and neet pg 14,870 views. This stains the decolorized cells pink and is less noticeable in the cells that still contain the crystal violet dye.
Staining techniques in microbiology archives learn. Some staining techniques, such as the gram and ziel neelsen stains, although. In addition to staining procedures, biochemical tests used for differentiating bacteria. Staining methods simple staining, negative staining, grams. Sterilization is the destruction or removal of all microorganisms, including bacteria and their endospores, viruses, fungi, and prions. 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.
Gramstaining is a differential staining technique that. Used to study small particles like viruses or macromolecules. When it is necessary to demonstrate whether or not a particular organism possesses a capsule, is flagellated, or forms endospores, special staining techniques must be used. A gramstaining technique combining staining with two fluorescent stains, oregon greenconjugated wheat germ agglutinin wga and hexidium iodide hi followed by flowcytometric detection is described.
Make a thin film of the material on a clean glass slide, using a sterile loop or swab for viscous specimens. Cover different classification schemes for grouping bacteria, especially the use of the gram stain 2. Techniques for controlling variability in gram staining of. Flood slide with crystal or gentian violet 60 seconds.
Sep 19, 2016 gram staining procedure animation microbiology principle, procedure, interpretation duration. Histologic staining options available include, but are not limited to. Some involve a single stain and just a few steps, while others use multiple stains. Types of different staining techniques of microorganisms. Gram staining helps to identify bacterial pathogens in specimens and cultures by their gram reaction grampositive and gramnegative and morphology coccirod. Staining and streaking protocols for simple stain, gram stain, streak plate technique and culture maintenance.
Under anaerobic conditions, grampositive staining occurred in all test organisms with quick fixing techniques with both absolute methanol and formalin. Staining is a supplementary method that gives divergence to the microscopic image for better vision under the microscope. Gram stain the previous lab introduced simple staining techniques that enable microbiologists to observe the morphological characteristics of bacteria. Staining is a technique where specimens are stained with specific dyes to create contrast between the. 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. Bacterial classification, structure and function introduction the purpose of this lecture is to introduce you to terminology used in microbiology. Pdf correctincorrect ratios in staining techniques in. Types of staining techniques used in microbiology and their. Hematoxylin has a deep bluepurple color and stains nucleic acids by a complex, incompletely understood reaction. 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. The wide application of sterilization processes makes it necessary to impose strict control measures to validate the resu lts.
Special stains some bacteria have characteristic surface structures such as capsules or flagella and internal components e. This can be accomplished by physical methods such as heat, radiation, and filtration, or by chemical methods. In the first step the smear is stained with basic dye crystal violet primary stain followed by treatment with iodine solution functioning as mordant. Learn staining techniques microbiology with free interactive flashcards.
Cell staining in microscopy types, techniques, preparations. Neelsen in 1883 used ziehls carbolfuchsin and heat then decolorized with an acid alcohol, and counter stained with methylene blue. The end result is a report that should assist the clinician in reaching a definitive diagnosis and a decision on antimicrobial therapy. Smears were then blocked with 1 % bovine serum albumin bsa in trisbuffered saline tbs with 0. Staining techniques in microbiology a presentation by g. Jun 12, 2018 it is the differential staining techniques which was first developed by ziehl and later on modified by neelsen. Feb 03, 2015 staining techniques in microbiology 1. Mcqs in microscopy and staining techniques part i category.
Gram staining in microbiology, fast staining in haematology, kit for staining reticulin fibres, pas staining and massons trichromic staining. Microbiology is a popular option for practical work in schools. Provide additional practice scenarios in staining techniques, with emphasis on tec hniques that require more than minute to apply a stain onto a smear e. 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. It is the differential staining techniques which was first developed by ziehl and later on modified by neelsen. Then apart from all this some other staining techniques are there for identification of fungus like lactophenol cotton blue staining, and many other techniques. Gram staining procedure animation microbiology principle, procedure, interpretation duration. Staining, therefore, is of primary importance for the. The gram stain is a differential stain commonly used in the microbiology laboratory that differentiates bacteria on the basis of their cell wall structure. In microbiology the concept of staining is very important because it highlights the structures of microorganisms allowing them to be seen under a microscope. The classic silver staining method for detecting bacterial lipopolysaccharides lpss in polyacrylamide gels c. The gram staining procedure is based on the fact that crystal violet stains the bacterial peptidoglycan cell envelope. Hans christian joachim gram the gram stain was devised by the danish physician, hans christian joachim gram, while working in berlin in 1883. Types of staining techniques used in microbiology and.