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The pour plate technique is the typical technique used to prepare plate count agars. Here, the inoculum is added to the molten agar before pouring the plate. The molten agar is cooled to about 45 degrees Celsius and is poured using a sterile method into a petri dish containing a specific diluted sample.
Colony-forming units are used to quantify results in many microbiological plating and counting methods, including: The pour plate method wherein the sample is suspended in a Petri dish using molten agar cooled to approximately 40–45 °C (just above the point of solidification to minimize heat-induced cell death).
The plates are left upright on the bench to dry before inversion and incubation at 37 °C for 18 – 24 hours (or appropriate incubation conditions considering the organism and agar used). Each sector is observed for growth, high concentrations will give a confluent growth over the area of the drop, or a large number of small/merged colonies.
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The inoculation loop is then dragged across the surface of the agar back and forth in a zigzag motion until approximately 30% of the plate has been covered. The loop then is re-sterilized and the plate is turned 90 degrees. Starting in the previously streaked section, the loop is dragged through it two to three times continuing the zigzag pattern.
Following the allotted time, the plate is removed and checked for bacterial growth. If the broth became cloudy or a layer of cells formed at the bottom, then bacterial growth has occurred. The results of the broth microdilution method are reported in Minimum Inhibitory Concentration (MIC), or the lowest concentration of antibiotics that stopped ...
The plate is removed from the container before the solvent reaches the top of the plate; otherwise, the results will be misleading. The solvent front, the highest mark the solvent has travelled along the plate, is marked. Visualization: The solvent evaporates from the plate. Visualization methods include UV light, staining, and many more.
The sample is then cooled and inspected for pour point as per the usual pour point method. The method usually gives higher pour point because the thermal history has not been cancelled by a prolonged thermal treatment. The lower pour point is measured by first pouring the sample into a stainless steel pressure vessel. The vessel is then screwed ...