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