It is a primary test which quickly and cost effectively divides bacteria into one of two types. Ellen jo baronlarry carlsoncarla clausensharon hilliargreg raugimichael rinaldijane schwebke edited and revised by rebecca buxton corresponding author department of pathology university of utah salt lake city, ut 842 usa email. Nov 03, 20 it includes introduction, history,principle, method of gram staining procedure slideshare uses cookies to improve functionality and performance, and to provide you with relevant advertising. The yellow contrast in the connective tissue collagen within skin tissue is. Module the gram stain procedure michigan state university. It was a few years later, that the german pathologist carl weigert18451904 from frankfurt, added a final step of staining with safranin. This video demonstrates the gram staining method for identifying unknown bacter. Sputum grams stain and culture can be performed noninvasively and. If flash is not working for you we have an alternate video version. Gram staining is the most sensitive method by which to visualize and recognize nocardiae in clinical specimens. The procedure is named for the person who developed the technique, danish bacteriologist hans christian gram. June 12, 2018 by sagar aryal 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. Flood airdried, heatfixed smear of cells for 1 minute with crystal violet staining reagent. The procedure for the gram stain was developed ppt.
Gently flood smear with crystal violet and let stand for 1 minute. Different bacteria stain differently to a common staining procedure. Gram positive and gram negative, based on the differences of the chemical and physical properties of the cell wall. Flood slide with crystal or gentian violet 60 seconds. Gram himself never used the red counterstaining in order to visualize the gram negative bacteria. Gram staining is the common, important, and most used differential staining techniques in microbiology, which was. June 12, 2018 by sagar aryal gram staining is the common, important, and most used differential staining techniques in microbiology, which was introduced by danish bacteriologist hans christian gram in. The two most important methods, the gram and acidfast techniques, use staining, decolorization, and counterstaining in a manner that helps to classify as well as stain the organism. Responsibility microbiologist or above of microbiology laboratory. Oct 24, 2019 both gram positive and gram negative bacteria pick up the pink stain, but it is not visible over the darker purple of the gram positive bacteria. Sputum gram s stain and culture can be performed noninvasively and are inexpensive diagnostic tests. Asmscience interpretation of gram stains and other common. Staining flood the entire slide when crystal violet, iodine and safranin are applied.
Gram variable observations in gram staining the gram staining procedure does not always give clearcut results. Sop on gram staining objective to lay down the procedure for gram staining. The gram stain procedure click start lab from a liquid culture, take a loopful of bacteria emulsify it in a small drop of water or saline on the slide. Alternatively, basic fuchsin can be used as the counterstain for weakly staining gramnegative organisms.
Bacteria may be stained by a variety of dyes, including. Gram staining procedure was discovered by hans christian gram in 1884. Well, thats where a gram staining procedure comes into play. Gram staining is a universal staining technique used for identification and classification of organisms. Asmscience interpretation of gram stains and other. Please note that the quality of the smear too heavy or too light cell concentration will affect the gram stain results.
Gently flood the smear with grams iodine and let stand for 1 minute. Gather all equipment and supplies gram stain presentation transcript. It includes introduction, history,principle, method of gram staining procedure slideshare uses cookies to improve functionality and performance, and to provide you with relevant advertising. Christian gram in 1884, and categorizes bacteria according to their gram character gram positive or gram negative. Sign in to view the content amrita vishwa vidyapeetham. It is one of the differential stains that are used to characterize bacteria in one of two groups. Gram staining is a quick procedure used to look for the presence of bacteria in tissue samples and to characterise bacteria as gram positive or gram negative, based on the chemical and physical properties of their cell. Bet you didnt know gram stains are performed on body fluid or biopsy when infection is presumed. On the other hand, some gramnegative bacteria may become grampositive in older cultures.
Corynebacterium diptheria seen in gram staining 35. Gramstaining chelsea bianchi how does gramstaing work. The procedure figure 1 involves the application of a solution of iodine in potassium iodide to cells previously stained with an acridine dye such as crystal violet. Gram staining procedure and its details general microscience. Hold the slide in a tilted downward position and allow the. A rapid method for distinction between gramnegative and grampositive bacteria by means of a 3% solution of potassium hydroxide is tested on 71 grampositive and 55 gramnegative bacterial strains. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Ppt gram staining powerpoint presentation, free download id. Cells without stain cheek cells with simple stain methylene blue simple stain. Carefully decolorize with 95% ethanol until thinnest parts of the smear are colorless. This procedure is based upon the hucker modification of the original grams stain method. Place slide with heat fixed smear on staining tray. Decolorization the critical step of the gram staining procedure is the decolorization step. Gram and acidfast stain control slides elitechgroup.
Find powerpoint presentations and slides using the power of, find free presentations research about gram staining ppt. This should be a thin, not milky, suspension or it will not stain properly. Gather all equipment and supplies gram stain microscope s heat source sterile swab culture or specimen microscope gram stain procedure 2. A rapid method for distinction between gram negative and grampositive bacteria by means of a 3% solution of potassium hydroxide is tested on 71 gram positive and 55 gram negative bacterial strains. Gram staining chelsea bianchi how does gram staing work. For this project, youll need a heatfixed bacteria slide, grams lodine, crystal violet, safranin, decolorizer, and water.
Gram staining is still the cornerstone of bacterial identification and taxonomic division. Trouble shooting in gram staining method the method and techniques used. Gram staining is a differential staining technique that provides an easy differentiation of bacteria into one of two groups. Here iodine favors more interaction between cell and the dye crystal violet. It includes introduction, history,principle, method of gram staining procedure. Schematic showing the modified gram stain procedure. Gather all equipment and supplies gram stain microscope slides heat source sterile swab culture or specimen microscope gram stain procedure 2. Principle, procedure, interpretation, examples and. Gram stain kits is used to stain microorganisms from cultures and specimens by the differential gram method. Young cultures must be young within 1824hrs old older cultures lose their gram staining properties due to changes in the cws as the cells get older thin smear thicker or uneven smears will result in uneven staining and decolorization fresh reagents of proper. In this staining, method bacteria are classified into two groups that are. Ppt gram stain procedure powerpoint presentation free.
The grams stain is used to classify bacteria on the basis of their forms, sizes, cellular morphologies, and gram reaction. How to perform a gram staining procedure wonderhowto. How to prepare an isolation streak plate powerpoint show or pdf printout printing. Steps in gram staining procedure follow the clock 1 on a rack, flood with filtered crystal violet 10 sec 2 wash briefly in water to remove excess crystal violet 3. Gram staining is a quick procedure used to look for the presence of bacteria in tissue samples and to characterise bacteria as grampositive or gramnegative, based on the chemical and physical properties of their cell walls. Apr 04, 2016 on the other hand, some gram negative bacteria may become gram positive in older cultures. Gently spread the bacteria on the slide use a circular motion. In addition this stain also allows determination of cell morphology, size, and arrangement. Interpretation of results if the gram stain procedure has been performed properly and the reagents are fresh, the staphylococcus aureus atcc 25923 in the positive control well should appear as a purple to lavender grampositive cocci. While still holding the inoculated swab, pick up your slide. Flood the slide with grams iodine and time for one. The traditional gram stain procedure is completed in either case top box.
Wash slide in a gentle and indirect stream of tap water for 2 seconds. This will ensure that all areas are stained evenly. Gram staining or gram stain, also called grams method, is a method of staining used to differentiate bacterial species into two large groups gram. If the staining procedure is performed correctly, gram positive bacteria will be purple, while gram negative bacteria will be pink. Gram positive bacteria will be purple and gram negative bacteria will be colorless. The best way for students to print out the powerpoint show is to download the pdf version. Jan, 2017 gram staining or gram stain, also called gram s method, is a method of staining used to differentiate bacterial species into two large groups gram positive and gram negative. Learners view video clips demonstrating grams staining procedure. The name comes from the danish bacteriologist hans christian gram, who developed the technique. View and download powerpoint presentations on gram staining ppt. Place slide on staining tray or hold with forceps above the sink. Oct 16, 2012 this video demonstrates the gram staining method for identifying unknown bacter. Qslide gram a microscope slide for the quality control of.
For this reason it is strongly recommended to use very young cultures for the staining procedure, after growth has become just visible. Principle, procedure, interpretation, examples and animation last updated. Gram stain or gram staining, also called grams method, is a method of staining used to distinguish and classify bacterial species into two large groups. Scope this sop is applicable for gram staining in microbiology laboratory of pharmaceutical company name. The name comes from its inventor hans christian gram who developed the technique in 1882 published 1884.
Their cell wall structure includes a thick peptidoglycan layer and teichoic acids. Gram stain is probably one of the most commonly used staining procedures used in the field of microbiology. The staining technique, developed in the late 1700s by christian gram classifies the rigid cell walled bacteria into one of two groups based on whether they are able to resist the decolorizing action of an alcoholic. Methylene blue, crystal violet, carbolfuchsin red, and safranin red. The gram stain is a differential method of staining used to assign bacteria to one of two groups grampositive and gramnegative based on the properties of their cell walls. Hold the slide in a tilted downward position and allow the decolorizer to flow over the smear. Gram staining gram staining procedure prepare a heat fixed smear of the culture you wish to examine flood the smear with crystal violet 30 sec.
Ppt gram stain powerpoint presentation free to view. Gram staining lab 3 hans christian gram developed gram staining in 1844 the reagents primary stain. Mordants are substance that improves the staining property of the dye. The procedure for the gram stain was developed in the 1880s and is still used to differentiate bacteria into two categories. There are 4 conditions to be followed for a valid gram staining procedure. Gram stain was described by danish bacteriologist hans christian gram in 1884 to differentiate. Gram did not actually use safranin as a counter stain in the original procedure gram negative cells would be colorless. A free powerpoint ppt presentation displayed as a flash slide show on id. Grampositive bacteria produces a bluepurple color with this staining procedure while gramnegative ones are red in color 5. Dotson md, jeanine wienerkronish md, in critical care secrets fourth edition, 2007. Ppt gram staining techniques powerpoint presentation, free.
Gram staining gram staining is the single most important and universally used staining technique in the laboratory. The name comes from the danish bacteriologist hans christian gram, who developed the technique gram staining differentiates bacteria by the chemical and physical properties of their cell walls. The gram stain should almost always be done as the first step in diagnosis of a bacteria infection the gram stain is named after the danish scientist hans christian. It is a critical test for rapid presumptive diagnosis. Gram staining method, the most important procedure in microbiology, was developed by danish physician hans christian gram in 1884. Tilt the slide slightly and gently rinse with tap water or distilled water using a wash bottle. Gram staining procedure a haiku deck by deidre holsten. This differential staining procedure separates most bacteria into two groups on the basis of cell wall composition. How to do a grams stain test grams stain is one of the most frequently used processes in identifying bacteria and is used daily in hospitals. Wash briefly in water, do not let the section dry out. Our gram stain control slides have two wells containing a gram positive control organisms staph.
Gram positive bacteria are cells that take up a purple color in the gram stain procedure. The gram stain of a bacteria is determined by one thing gram positive gram negative the thickness of the petidoglycan in its cell wall materials background clothes pin 2. Ppt gram stain procedure powerpoint presentation free to. Gram himself never used the red counterstaining in order to. Flood the surface of the slide with crystal violet stain and let sit for one minute. Beyer individual supplies microscope slide clothes pin inoculating loop or wood splint paper towels shared supplies crystal violet stain safranin red stain grams iodine ethyl alcohol denatured water bottle rinsing basin alcohol burner wax pencil matches microscope create a staining area on your microscope slide obtain a clean microscope slide draw a. Overheating of smears when heat fixation is used d. The gram stain control slides are prepared from standard quality control organisms.
Bacteria are translucent staining make them visible under microscope 1884 hans christian gram stained cells and. An optimized staining technique for the detection of gram. Gram stain or gram staining, also called gram s method, is a method of staining used to distinguish and classify bacterial species into two large groups. Learning how to perform a gram stain will be the starting step in determining where to start when identifying bacteria for illnesses, disease or infection.
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