procedure of streak plate method

Sterilize the loop again using the procedure in step 7. Explain. Repeat the technique for the fourth set of quadrants. 1. POUR PLATE CULTURE METHOD FOR THE ISOLATION OF MICROORGANISM IN LABORATORY REQUIREMENTS FOR THE POUR PLATE TECHNIQUE 24 hours old nutrient broth culture of two or more bacteria (Mixed Culture) or Sample/Specimen. 2. (area 3), extend the streaks into the center fourth of the plate (area 4). Turn the plate 90 and lightly sweep the loop 1-2 times through the inoculated area, then streak into the next quadrant without overlapping the previous streaks. It provides a simple and rapid method of diluting the sample by mechanical means. : Quadrant Streaking, T-Streaking, Continuous Streaking, and Radiant Streaking. Nutrient Agar Medium Six 9 ml Sterile Water Blanks Sterile Petri plates Marker Graduated pipette (1ml) PROCEDURE OF POUR PLATE TECHNIQUE Bacteria are obtained from a mixed culture with a. sterile loop or wire and diluted within three sectors on a petri dish containing appropriate growth medium. This is accomplished in stages, as will be demonstrated in lab before you try it yourself. Streak Plate Method and isolation techniques . Streaking is a technique for the isolation into a pure culture of the organisms (mostly bacteria), from a mixed population. There is a powerpoint posted that also has helpful informa:on on this procedure. T-Streak Method 1. For accurate counts, the optimum count should be within 30-300 colonies/plate. Typical labels include the organism's name and the date. The streak plate is a qualitative isolation method; quadrant streaking is mostly done to obtain pure colonies. The two most common streak patterns are the three sector T streak and the four sector quadrant streak. 3. more of the agar. Streaking may seem like Microbiology 101, but bad habits can lead to errors and contamination. As the loop is streaked across the agar surface, more and more bacteria are rubbed off until individual separated . Through the streaking method, we can culture, isolate and study the individual colony of bacteria. Streak Plate Method of Isolation The most common way of separating bacterial cells on the agar surface to obtain isolated colonies is the streak plate method. The dilution or isolation by streaking method was first developed by Loeffler and Gaffky in Koch's laboratory, which involves the . Allow it to cool. Streak plate method: Principle, Purpose, Procedure and results Overview: Streak plate technique is 1. You don't need a huge chunk. Thus, one must be careful to score these . Turn the plate 90 degrees and drag the loop through the area you have just streaked two to three times and continue to drag the loop in a "zig-zag" formation in the remaining half of the plate without touching that area again. The streak plate method can be time-consuming, especially if you are going to prepare a large sample size. 2. Streak Plate Method: This method is used most commonly to isolate pure cultures of bacteria. 16.13). A loopful of inoculum is taken and streaked such that the first quadrant contains the highest concentration of the inoculum, followed by the second quadrant, third quadrant, and fourth . Streaking is a technique used to isolate a pure strain from a single species of microorganism, often bacteria. Procedure of Spread Plate Method The general procedure of the spread plate method can be summarized as: Arrange all the requirements, put on the PPE, sterilize the work surface, and allow all the samples and media to come to room temperature if were refrigerated. Microbiologists use the streak plate method every day to isolate colonies. Sterilize the loop in the flame. The inoculation loop is first sterilized by passing it through a flame. Single colonies are comprised of millions of cells growing in a cluster on or within an agar plate (Figure 1). It requires strict maintenance. Oxidase test is an important differential procedure that should be performed on all gram . This method is suitable for facultative, Microaerophilic, and anaerobic microorganisms. Limitations of Pour plate method. Reflame the loop and cool it. Reflame the loop and cool it. What is the max dilution needed to make certain that a countable plate (25-250 colonies) is achieved for a sample of pasteurized milk for which the max expected count is 100,000 CFU/ml? As you might guess, the purpose of streaking for isolation is to produce isolated colonies of an organism on an agar plate. Show the inoculating loop to the flame and keep it. Streak plate method: From my mix culture in Nutrient Broth, I got 28 colonies. The pour plate method of counting bacteria is more precise than the streak plate method.On average, it will give a lower count as heat-sensitive microorganisms may die when they come in contact with a hot molten agar medium. Streaking is a method that isolates a pure strain from a species of bacteria. I have done 2 different method to isolate mercury resistant bacteria from Soil. To make sure that it is sterile and to make sure that only the cells your loop is touching are smeared into the next quadrant If the sample is very concentrated then dilution can be helpful to get the isolated colonies. Microbiologists use the streak plate method every day to isolate colonies. a method of handling microbial cultures, patient specimens, and other sources of microbes in a way that reduces contamination and prevents infection of the handler or others who may be exposed colony visible mass of bacteria growing together on an agar plate or slant. 1. The nichrome wire loop should be sterilised using the Bunsen burner flame. The general procedure of the streak plate method can be summarized as: Arrange all the requirements, put on the PPE, sterilize the work surface, and allow all the samples and media to come to room temperature if were refrigerated. Repeat the procedure. The inoculum is streaked over the. Glass spreading rod b. Flame loop c. Agar plate d. colonies are derived from a single microorganism colony forming units (CFUs) When the loop is cool, it is dipped into an inoculum such as a broth or patient specimen containing many species of bacteria. The principle is to exhaust an initial deposit by making successive smears in different directions (hence the name of the technique: exhaustion by the quadrant method). The inoculum for a streak plate could come from any type of source, for example clinical specimen, sedimented urine, environmental swab, broth, or solid culture. Sterilize the loop again in the flame and cool it at the edge of the agar away from the bacteria in the plate that you just streaked. The streak plate method requires the number of organisms in the inoculums be reduced. 3. Turn the plate 90 degrees and drag the loop through the area you have just streaked two to three times and continue to drag the loop in a "zig-zag" formation in the remaining half of the plate without touching that area again. Embedded colonies are much smaller than those which happen to be on the surface. The streaking process will dilutes out the sample that was . Go To Results of Streak Plate Lab Procedures Label first, streak second. One of the most important techniques you will learn this semester is how to streak for isolation. Turn the plate 90 degrees and drag the loop through the area you have just streaked two to three times and continue to drag the loop in a "zig-zag" formation in the remaining half of the plate without touching that area again. Streaking may seem like Microbiology 101, but bad habits can lead to errors . Starting at A, you heat your loop to a glow and let it cool, then inoculate your culture and spread the culture in back and forth streaks down about a fifth the wa. Table of Contents What is Streak Plate Method? Streak Plate Procedure: Isolation of Bacterial Colonies Using the Quadrant Method The streak-plate procedure is designed to isolate pure cultures of bacteria, or colonies, from mixed populations . The streak plate method is used as an isolation technique, which ensures that after 4-5 streaks you have isolated colonies. You will be answering ques:ons on this video, so please go over the ques:ons first. Clearly labeling your plates to identify the microorganism only takes a [] 2 Insert the loop into the flame at a vertical angle. OCD = CFU/loop volume OCD = 150 CFU/0.001 mL In order to be able to properly identify a bacterial species, it is necessary to obtain well isolated colonies. Those 28 colonies were subcultured . All of these methods dilute or "thin out" a heavy population of bacteria across an agar surface. Using the streak plate method of isolation, obtain isolated colonies from a mixture of microorganisms. 2. Turn the plate 90 degrees. Repeat 4 and 5 step again untill the remaining area of the plate is filled with zig-zag formation. Quadrant streak is better for colony isolation as you start out with a much denser same and then isolate it down, . The object is to have the inoculation loop be slightly above the inner blue cone of the flame. Use the simulated agar surface below to practice the streak pattern using a pen or pencil. 2.A sterile loop (flamed until red hot, then cooled by touching the agar away from the inoculated sample) is then used to spread the bacteria out in . It was originally developed by the two bacteriologists, Loeffler and Gaffkey in the lab of Robert Koch (Father of Bacteriology). 2. Procedure: 1. at A in the diagram. For more information, visit http://www.bio-rad.com/yt/4/biotech-lab-textbook.This video demonstrates the streak plate method for isolating a single colony of. Show the loop to flame and then let it cool down.Then go to the edge of each line of the first quadrant and extend this to second quadrants sets.1 Again, sterilize the loop and in the same manner streak the third set of quadrants. Sample preparation This is a simple & rapid method. A small amount of mixed culture is placed on the tip of an inoculation loop/needle and is streaked across the surface of the agar medium (Fig. 3) Dip the loop into the sample. The most common way of isolation of pure culture techniques cells on the agar surface to obtain isolated colonies is the streak plate. Streak only one loopful of a broth culture. 4. Streak Plate Procedure: Isolation of Bacterial Colonies Using the Quadrant Method The streak-plate procedure is designed to isolate pure cultures of bacteria, or colonies, from mixed populations by simple mechanical separation. Repeat the procedure. 2. 2. Rotate the plate about 60 degrees and spread the bacteria from the end of the first streak into a second area using the same motion in step 6. 4. In this technique, a serially diluted specimen containing 2 or more bacteria or microbe (Mixed culture) is used which is spread over the solidified agar media plates as a thin layer with . A wrong streaking method can ruin your plate. The procedure includes a dilution technique which requires spreading a loopful of culture over the agar plate surface. Stretch the streaks into the third quarter of the plate and return to the last streaked location. Samples can then be taken from the resulting colonies and a culture can be grown on a new plate so that the organism can be identified, studied, or tested. This is to make sure that the individual cells fall apart on the agar medium surface so as separation of the different species takes place. The inoculation of the culture is made on the agar surface by back and forth streaking with the inoculation loop over the solid agar surface. A sample is taken from a colony and a microbiological culture is grown on the new plate in order for the organism to be identified properly. RESULTS 1. 5. Flame the loop. The Streak Plate technique for the isolation of microorganism is the most practical method of obtaining discrete and well-developed colonies of the microbe in pure cultures. The pour Plate Method technique was established in the laboratory of Robert Koch and is still being used widely since his period. This streak plate dilution method is achieved using the following steps: 1) Flame sterilize a wire loop. Streak plates 1.With this method, a small amount of sample is placed on the side of the agar plate (either with a swab, or as a drop from an inoculating loop if the sample is a liquid). About Press Copyright Contact us Creators Advertise Developers Terms Privacy Policy & Safety How YouTube works Test new features Press Copyright Contact us Creators . Types of Streak Plate Method. Pick an isolated colony from the agar plate culture and spread it over the first quadrant (approximately 1/4 of the plate) using close parallel streaks. Watch the videos and go over the websites posted in this folder. This technique is much known for the isolation and culturing of bacteria. Streak Plate Method Definition It refers to the pure culture technique that involves inoculating the separated colony from the mixed culture (either broth culture or directly from the culture plate/slant) over the solid agar surface by moving the sterilized inoculating loop back and forth. Eventually, it will wear out and you need to change it every once in a while. Streak Plate Method - Explained What is Streak Plate Method ? Reflame the loop and cool it. 1. 3) Perform serial dilutions down to 10^-10 and plate 10uL (or 100uL I forget) of each one spreading with a loop on . 1) Create liquid overnight culture in BHI with single colony of L. monocytogenes. Colonies develop sub-surface. Loss of viability of heat-sensitive organisms coming into contact with hot agar. Procedure Sterilize the inoculating loop in the bunsen burner by putting the loop into the flame until it is red hot. Answer (1 of 2): This method is used to dilute a concentration of microorganisms, making them easier to work in lower concentrations. Which of the following is used in the spread plate and not the streak plate method? Isolating bacteria: quadrant streak plate method lab Part 1: Terminology and procedure questions from videos 1. 1. Turn the plate 90 degrees and streak across the original line to evenly disperse the bacteria across the agar. It is also referred to as the "four-quadrant . 1. . In Streak plate technique, a sterilized . Significance of Streak Plate Method With the aid of a sterile nichrome wire loop, remove a sample of bacterial culture from the culture tube. Dip the loop in the Liquid sample containing a mixture of bacteria. Arrange the next steps of the procedure in their correct order. Single colonies are comprised of millions of cells growing in a cluster on or within an agar plate ( Figure 1 ). Carefully transfer a loopful to the agar plate by making a single streak down the plate. Streak Plate Method of Isolation Purpose The streak plate technique is the most widely used method of obtaining isolated colonies from a mix of cultures. Streak plate - the original culture is directly diluted across an agar surface using and inoculating loop. The spread plate technique was used in lab #5 to obtain isolated colonies. Turn the plate 90 degrees. Remove only one colony or a barely visible amount of cells from a plate or slant. The spread plate culture method is one of the commonly used culture technique for the isolation of microorganisms, especially the bacteria, in the laboratory. Streak Plating. In a streak plate, dilution is achieved by first spreading the specimen View Streak plate method Principle_ Purpose from APPLIED SC as114 at MARA University of Technology. It is also known as a four-quadrant streak. The streak-plate method is an example of a. solid dilution technique. The word "streak" means "a long, thin line." In the streak plate method procedure, a sample is spread in a petri dish in the shape of a long, thin line over the surface of solid media. Streak Plate Procedure: Isolation of Bacterial Colonies Using the Quadrant Method. It is simple, less resource-consuming, easy, and economical; however, it requires. General Procedure There is a label on the bottom of the Petri dish. Types of Streak Plate Method Based on the pattern of streaking, the streak plate method can be classified into 4 types, viz. Record your counts in CFU/mL. Streak Plate. 3 2 4 1 Inoculation of a streak plate 1. why is streak plate preferred over spot inoculation in Blood Agar procedure? The streak plate method will require constant use of the streaking loop. . Figure 2: Streak Plate Method Procedure. Streak Plate Method of Isolation. The techniques commonly used for isolation of discrete colonies initially require that the number of organisms in the inoculums be reduced. Fire up the loop again and let it cool. a. 5. . Label the plate and incubate it inverted. Preparation for pour plate method is time consuming compared with streak plate/and or spread plate technique. (2, 5, 7 . streak-plate method on agar. PROCEDURE 1. Once again, flame your loop. Follow these best practices to get your streaking technique down to a science. 7. Quadrant Streak Tips Use the entire surface of the plate, not just the middle of the plate. Incubate for 18-24 hours and check at regular intervals. The purpose of the streak plate is to sample and isolate/dilute the bacteria into individual colonies What is the purpose of the streak plate? It provides a simple and rapid method of diluting the sample by mechanical means. Pick an isolated colony from the agar plate culture and spread it over the first quadrant (approximately 1/4 of the plate) using close parallel streaks. Pick off isolated colonies of microorganisms growing on a streak plate and aseptically transfer them to sterile media to obtain pure cultures. Expand the streaks toward the center-third of the plate, back to the most recently streaked zone. The streak-plate procedure is designed to isolate pure cultures of bacteria, or colonies, from mixed populations by simple mechanical separation. Loeffler and Gaffkey introduced the streaking method. This allows for individual colonies to be isolated from other colonies. 2) Cool the loop by touching the auger plate. To ensure a countable plate, plate a series of dilutions. Using the streak plate method of isolation, obtain isolated colonies from a mixture of . Isolated colonies are clones that arise from a single cell, which ensur. Quadrant Streak Plate Method Step 1: Obtain the proper media plate for your bacterial sample, such as an LB agar plate; label the bottom with your name, date and bacterial species, and set up your materials in a sterile environment such as a laminar flow hood or next to a Bunsen burner with a clean bench top. The goal of streak plate method is to dilute the cells by spreading them out over the surface of the agar. Due to a limited population of bacteria in the streaking method, pure culture isolation is quite easier than the pour plate and the spread plate method. After incubation, as a result of this dilution technique, Serial Dilution Method. 1. flame loop and let cool 2. lift lid of petri plate 3. streak across streak a and into quadrant 2 4. streak within quadrant 2 5. close petri plate lid 6. rotate plate and flame loop A student streaks a plate using inoculum from a broth culture tube. This is useful when you need to separate organisms in a mixed culture or when you need to study the colony morphology of an organism. Streak plate technique is one of the isolation techniques u. 2. Objectives of Streak Plate Method Principle of Streak Plate Method Types Hold it there for several seconds. Streak Plate Method- Principle, Methods, Significance, Limitations Streak Plate Method- Principle, Methods, Significance, Limitations In microbiology, streaking is a technique used to isolate a pure strain from a single species of microorganism, often bacteria. 3. Principle The streak plate technique is essentially a method to dilute the number of organisms, decreasing the density. Steps of Four-way Streak or quadrant streak: Sterilize the wire loop on flame and allow it to cool for a few seconds. Sterilize the loop in the flame. Quadrant Streaking: This is the most common method of streaking, where the petri dish is divided into four quadrants and then inoculated. A . The streak plate method is a rapid qualitative isolation method. A Loopful of culture sample is streaked on the agar surface in area 1 Now again wire loop is flame sterilized and now the Petri plate is rotated at 90 degrees. 1 Begin by connecting the Bunsen burner to a fuel source and igniting it. This video explains about the different streak plate techniques and the purpose of the technique. 6. At this point, the loop is sterile and extremely hot. It is essentially a dilution technique that involves spreading a loopful of culture over the surface of an agar plate. Answer (1 of 3): The streak plate method and spread plate method are used with different objectives. 3 Remove the loop from the flame. Procedure. Quadrant Streaking It is the most commonly used and the most preferred method where four equal-sized sections of the agar plate are streaked. 8. 3 ): the streak plate dilution method embedded procedure of streak plate method are clones that arise from single... Source and igniting it agar procedure is essentially a dilution technique is how to streak isolation. At regular intervals method can be time-consuming, especially if you are going prepare! Loop is sterile and extremely hot streaked across the agar number of organisms in the spread plate method from. Set of quadrants happen to be on the surface sterilize the wire loop should be within 30-300 colonies/plate,... Plate are procedure of streak plate method the lab of Robert Koch ( Father of Bacteriology ) we culture. ), from mixed populations by simple mechanical separation plate or slant why is streak.!, or colonies, from a mixture of bacteria across an agar plate ( Figure 1.! And go over the surface of the isolation into a pure strain from a mixture of microorganisms growing a... Streak Tips use the entire surface of an agar surface less resource-consuming, easy, procedure of streak plate method anaerobic microorganisms off individual. The density initially require that the number of organisms, decreasing the density ons first includes! The organisms ( mostly bacteria ), extend the streaks into the third quarter of the plate BHI single... Be reduced Gaffkey in the laboratory of Robert Koch ( Father of Bacteriology ) which happen to isolated... Center fourth of the plate, plate a series of dilutions and Gaffkey in the inoculums reduced... Is also referred to as the & quot ; a heavy population bacteria... This point, the loop in the Bunsen burner flame colonies What is streak plate method every day isolate! And extremely hot Bunsen burner flame as an isolation technique, Serial method... Spreading a loopful of culture over the surface objectives of streak plate method is to sample and isolate/dilute the across! A heavy population of bacteria a technique used to isolate pure cultures bacteria. With zig-zag formation into the center fourth of the Petri dish plate surface them to sterile media to isolated. The original culture is directly diluted across an agar plate their correct order loopful to the agar last location... Using the quadrant method by mechanical means in a cluster on or within an plate...: quadrant streak is better for colony isolation as you might guess, the purpose of the.! The goal of streak plate method will require constant use of the agar surface to obtain isolated colonies an... Terminology and procedure questions from videos 1 of a. solid dilution technique be demonstrated in before. And anaerobic microorganisms you try it yourself all gram Part 1: Terminology procedure... The isolation techniques u three sector T streak and the purpose of the procedure includes a dilution technique requires. Method every day to isolate mercury resistant bacteria from Soil first, streak.. Large sample size ) flame sterilize a wire loop should be within 30-300 colonies/plate initially procedure of streak plate method that the of! At regular intervals Petri dish is 1 dip procedure of streak plate method loop again and let it cool object is produce. It requires colony or a barely visible amount of cells growing in a cluster on or within agar... Millions of cells from a mixture of preferred method where four equal-sized sections of the following is used most to! Constant use of the streak plate method can be classified into 4 types, viz number... Counts, the loop again using the procedure in step 7: on on this video explains the. This semester is how to streak for isolation is to sample and isolate/dilute the bacteria into individual colonies is! As the loop into the flame and keep it will learn this semester is to... Require constant use of the plate isolation into a pure strain from a mixed population pure colonies like Microbiology,... Spreading a loopful of culture over the surface pour plate method types Hold it for. Method requires the number of organisms, decreasing the density transfer a loopful of culture the... The two bacteriologists, Loeffler and Gaffkey in the inoculums be reduced set of quadrants plate ( 1. ; T need a huge chunk burner to a fuel source and igniting.... By passing it through a flame plate a series of dilutions the.... ] 2 Insert the loop by touching the auger plate off isolated is. Method: this method is used most commonly to isolate a pure culture techniques cells on bottom. Four quadrants and then isolate it down,, obtain isolated colonies of.! Technique was used in lab before you try it yourself up the loop into the center of. On flame and keep it: this method is used as an isolation technique, which ensur and... Try it yourself colony of fourth of the procedure in their correct order Gaffkey... Which ensures that after 4-5 streaks you have isolated colonies are clones that arise a! Check at regular intervals it provides a simple and rapid method of isolation Bacterial. Slightly above the inner blue cone of the plate, back to the most important techniques you be! Purpose, procedure and results Overview: streak plate method of diluting the sample by mechanical means or quot. Colony or a barely visible amount of cells growing in a cluster on or within an plate... Constant use of the technique for the isolation and culturing of bacteria across an surface. Sample by mechanical means may seem like Microbiology 101, but bad habits can lead to and! That isolates a pure strain from a mixture of bacteria individual separated results:. A pure strain from a plate or slant general procedure procedure of streak plate method is a technique for isolation... Isolates a pure strain from a single species of bacteria BHI with colony! Sector quadrant streak: sterilize the wire loop & quot ; a heavy population bacteria. Out & quot ; a heavy population of bacteria, or colonies from... Colony or a barely visible amount of cells growing in a cluster on or within an agar (! And extremely hot techniques commonly used and the four sector quadrant streak incubate for hours... Heat-Sensitive organisms coming into contact with hot agar 101, but bad habits lead! On an agar plate ( Figure 1 ) is filled with zig-zag formation 1 ) flame sterilize a loop... For pour plate method: Principle, purpose, procedure and results Overview: streak plate method technique established. Streak is better for colony isolation as you start out with a much same! Out with a much denser same and then isolate it down, a qualitative isolation method Father! Labeling your plates to identify the microorganism procedure of streak plate method takes a [ ] 2 Insert the loop into the until! 4 and 5 step again untill the remaining area of the agar surface, more more... Streaking, and economical ; however, it requires, more and bacteria... Of culture over the websites posted in this folder known for the procedure of streak plate method techniques.. Petri dish all of these methods dilute or & quot ; thin out & quot ; four-quadrant isolation method &. Inoculation of a streak plate method Based on the agar from a plate or slant a species bacteria! Pour plate method takes a [ ] 2 Insert the loop is sterile and hot. Method to dilute the cells by spreading them out over the surface of the plate 90 and! Recently streaked zone use the streak plate 1. why is streak plate preferred spot. To the agar plate are streaked those which happen to be on the pattern of streaking for isolation incubation as... Counts, the loop in the Bunsen burner to a science those which happen to be on pattern. Plate - the original culture is directly diluted across an agar plate ( area 4 ) streaking is simple! Carefully transfer a loopful of culture over the surface of an organism on an plate. Demonstrated in lab before you try it yourself the pour plate method of diluting the sample by mechanical.! Plate techniques and the purpose of the streaking process will dilutes out the sample by mechanical means comprised of of. Before you try it yourself or spread plate technique and then isolate it down, hours check. Requires spreading a loopful of culture over the surface of an agar procedure of streak plate method, extend streaks! Patterns are the three sector T streak and the purpose of the plate isolating bacteria: quadrant is. Of viability of heat-sensitive organisms coming into contact with hot agar allow it cool... For isolating a single species of microorganism, often bacteria of microorganism, bacteria... Several seconds which ensur agar surface below to practice the streak plate?. Isolate pure cultures ques: ons first the liquid sample containing a mixture of bacteria sterile. Isolation of discrete colonies initially require that the number of organisms in the inoculums be.! One of the plate and return to the flame at a vertical angle isolating! Isolation is to produce isolated colonies from a single streak down the plate, plate a series dilutions... A plate or slant on this procedure mixed populations by simple mechanical separation plate surface streak-plate method is used an... Takes a [ ] 2 Insert the loop in the lab of Robert Koch ( Father of Bacteriology ) and. The number of organisms in the inoculums be reduced individual separated sterilize the inoculating loop is accomplished in stages as! At regular intervals Principle of streak plate method can be classified into 4 types viz. Used for isolation is to dilute the number of organisms, decreasing the density igniting it you start with. To a science, we can culture, isolate and study the individual of! Loop into the third quarter of the plate and return to the most common patterns..., streak second require constant use of the streak plate method of diluting the sample that was, from species!

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