precautions of streak plate method

a method of isolating a culture by spreading microorganisms over the surface of a solid culture medium. 1 Comment. During streak ing, the cell density decreases, eventually leading to individual cells being deposited separately on the agar surface. If the plates are stored at 4 C, remove them several hours or even the day before. STREAK PLATE CULTURE TECHNIQUE FOR THE ISOLATION OF MICROORGANISM / BACTERIA IN PURE CULTURE. April 5, 2018. : Quadrant Streaking, T-Streaking, Continuous Streaking, and Radiant Streaking. 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. Accurately measure the quantity while preparing the serial dilutions of the specimen. Flame the loop again and allow it to cool. Now Streak the area 3 in the same manner as area 2, hitting the last area several times. PRECAUTIONS TO BE TAKEN.. The protocol should be followed under all aseptic conditions preferably in Laminar Air Flow (Safety cabinet) to avoid any contamination. Flame your loop once more. April 7, 2018. It is also referred to as the "four-quadrant . The streak plate method is a rapid and simple technique of mechanically diluting a relatively large concentration of microorganisms to a small, scattered population of cells. 1.Heat an inoculate loop to red-hot and then allow to cool for at least 5 seconds. Use the simulated agar surface below to practice the streak pattern using a pen or pencil. Precautions during Spread Plate Technique Follow proper safety protocols. Draw a letter T on the bottom side of an agar plate. One of the most important techniques you will learn this semester is how to streak for isolation. SAHIL BATRA. 11 Pour Plate Method Best Practices Keep the molten agar in the water bath for no more than three to four hours. 4. Accurately measure the quantity while preparing the serial dilutions of the specimen. The water or broth used in serial dilution must be sterile. Continuous Streaking 4. Streak the Plate: The lid of the agar plate has to be opened just sufficiently enough to streak the plate with the inoculation loop. 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. The spread plate method is a technique to plate a liquid sample containing bacteria so that the bacteria are easy to count and isolate. 1)streak q1 then flame loop. Reflame and Cool the loop and turn the Media plate to 90. 5. 2. Flame the loop again and allow it to cool. Cells that have been sufficiently isolated will grow into . The goal is to obtain isolated colonies on a large part of the agar surface, so that desired species can then be brought into pure culture. : Quadrant Streaking, T-Streaking, Continuous Streaking, Radiant Streaking, Semi-quantitative Streaking, Zigzag Streaking. Fire up the loop again and let it cool. isolate pure cultures. Quadrant Streaking It is the most commonly used and the most preferred method where four equal-sized sections of the agar plate are streaked. Make sure the agar plates are free of droplets of condensed moisture. Flame sterilize wire loop, allow it to cool completely, take loopful of sample and place in area 1 and streak back and forth in a zigzag motion to spread culture. Precautions Some preventive measures should be considered while performing the experiment: First, you should sterilize all the equipment. This is accomplished in stages, as will be demonstrated in lab before you try it yourself. Streak Plate Method of Isolation. Flame the glass spreader (hockey stick) over a Bunsen burner. A variety of techniques has been developed for the isolation of microorganism, mainly the bacteria, from. Quadrant Streaking 2. The goal of streak plate method is to dilute the cells by spreading them out over the surface of the agar. Stretch the streaks into the third quarter of the plate and return to the last streaked location. Zigzag Streaking Requirements of Streak Plate Method Procedure or Protocol of Streak Plate Method Quadrant Streaking Procedure Dip the L-shaped glass spreader into alcohol. Microbiology - 004 - Spread Plate Method. Make sure you streak lightly so as not to gouge the agar. The plates must be completely dry without condensation on the lid and pre-warmed to room temperature prior to streak-plating. Make sure that all the tools and glass wares are sterile. In the streak plate method of isolation, a bacterial sample (always assumed to be a mixed culture) is streaked over the surface of a plated agar medium. Proper streaking of plates is . plate-type heat exchanger (1) or an external wall (5) of the apparatus. Use phosphate buffer pH 7.2 if necessary to dilute the suspension. Treat every unknown or clinical specimen as hazardous and follow safety accordingly. 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. Minimize the amount of agar and the length of time the agar is exposed to the environment during the streak process. Three Sector Streak (t streak): Sterilize the wire loop. Going back to the area that you just streaked (area 2), extend the streaks into the third quarter of the plate (area 3). The resulting . 1. Precautions Of Pour Plate Method The protocol should be followed under all aseptic conditions preferably in Laminar Air Flow (Safety cabinet) to avoid any contamination. Expand the streaks toward the center-third of the plate, back to the most recently streaked zone. Radiant Streaking 5. 1. Microbiologists use the streak plate method every day to isolate colonies. Allow the media plates to uniformly solidify. After streaking each quadrant, do not forget to flame the loop. The agar media must be appropriately solidified, or it must be free of condensed moisture. 2 Comments. The inoculum source can be a broth or solid culture, an environmental swab, a clinical specimen, or sedimented urine. The techniques commonly used for isolation of discrete colonies initially require that the number of organisms in the inoculums be reduced. See to it that the plate's surface is free of droplets of condensed moisture. 3) spread from q2 to q3. Based on the pattern of streaking, the streak plate method can be classified into 6 types, viz. Follow these best practices to get your streaking technique down to a science. It would be best to label the culture plate before streaking to avoid the unwanted mess. Pipette out 0.1 ml from the appropriate desired dilution series onto the center of the surface of an agar plate. To ensure a countable plate, plate a series of dilutions. Label first, streak second. The measured temperature is compared with a temperature target value stored in a control device (30) of a plant, where the temperature target value is measured from rigidity . Significance of Streak Plate Method Immediately streak the inoculating loop gently over a quarter of the plate using a back-and-forth motion (see area 1 in the figure above). Streaking may seem like Microbiology 101, but bad habits can lead to errors and contamination. Don't re-melt the agar. Decrease the risk of contamination by pouring plates in a laminar-airflow cabinet. The method involves measuring temperature (T1-T3) in local areas (23-25) within an apparatus i.e. optimal result of streak plate method. For accurate counts, the optimum count should be within 30-300 colonies/plate. It is essentially a dilution technique that involves spreading a loopful of culture over the surface of an agar plate. Number three sections of the plate as 1,2,3. As you might guess, the purpose of streaking for isolation is to produce isolated colonies of an organism on an agar plate. 1. Reflame and cool the loop and turn the media plate 90 anti-clockwise. Do not. Keep the agar dry. Flame the loop. Follow these best practices to get your streaking technique down to a science. Allow the media plated to uniformly solidify. Make a dilution series from a sample. Procedure of Spread Plate Technique. Types of Streak Plate Method Based on the pattern of streaking, the streak plate method can be classified into 4 types, viz. Flame the loop again and allow it to cool. T Streak Steps: Take a sterile nutrient agar plate. T-Streaking 3. An inoculated plate is always incubated in an inverted position to prevent condensation from falling onto the surface of the plate and interfering with discrete colony formation. Spread them out in small, staggered stacks of no more than 2-3 plates and allow them to dry. 1. 3. Remove the cap of the bacterial culture tube and flame the neck of the tube, then remove a loopful of the culture using the sterilized inoculating loop. Accurately measure the quantity while preparing the serial dilutions of the specimen. 2. 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. Clearly labeling your plates to identify the microorganism only takes a few seconds and will help you avoid a big headache later. Going back to the area that you just streaked (area 3), extend the streaks into the center fourth of the plate (area 4). Touch the loop to the corner of the culture streak in Area 3 and streak the inoculum across the agar. Once again, flame your loop. Four sections of the agar plate of the same size are streaked. How do the colonies on the surface of the pour plate? SAHIL BATRA. 4)spread from q3 to. Spread the sample evenly over the surface of agar . SPREAD PLATE CULTURE TECHNIQUE FOR THE ISOLATION OF MICROORGANISM / BACTERIA IN PURE CULTURE. PRECAUTIONS TO BE TAKEN.. The protocol should be followed under all aseptic conditions preferably in Laminar Air Flow (Safety cabinet) to avoid any contamination. A successful spread plate will have a countable number of isolated bacterial colonies evenly distributed on the plate. Returning to the edge of area 1 that you just streaked, extend the streaks into the second quarter of the plate (area 2). Semi-quantitative Streaking 6. The streak plate method is a rapid qualitative isolation method. Use the sterile pipettes each time to avoid any contamination and errors in the result. Objectives of Streak Plate Method Principle of Streak Plate Method Types of Streak Plate Method 1. What precautions should be taken when the pour plates? Quadrant streak B method. 2. A variety of techniques has been developed for the isolation of microorganism, mainly the bacteria, from. 2)spread q1 to q2 and reflame. Purpose of streak plate method. 2. Quadrant Streaking It is the most common and preferred method. List the basic steps of the streak plate method in the correct order from start to finish. 3. Practices Keep the molten agar in the water or broth used in serial dilution must sterile! 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