standard plate count method microbiologymike gundy mullet 2019
(Use vacuum to hold microscope cover slip against face of stylus tip; if cover slip plane is parallel at about l mm from surface of platform, tip is properly oriented). Place agar plate on platform, place stylus tip on agar surface, and start motor. After incubation, center spiral plate over grid by adjusting holding arms on viewer. In this case, count each circumferentially adjacent segment in all 8 wedges, counting at least 50 colonies, e.g., if the first 2 segments of a wedge contain 19 colonies and the third segment contains the 20th and 76th (or more), count colonies in all circumferentially adjacent first and second segments in all 8 wedges. Complete by counting remaining colonies in segment where 20th colony occurs.
Aseptically transfer 100 μL from tube #6 to tube #7, close the lid and vortex.A turbid broth tube contains millions of bacteria. If we transferred directly from that tube, we would get a plate that had a lawn of bacteria. From that number, we can calculate the original cell density in the broth.3. They should not have water droplets on surface of agar or differences greater than 2 mm in agar depth, and they should not be stored at 0-4.4°C for longer than l month. Clean stylus tip by rinsing for 1 s with sodium hypochlorite solution followed by sterile dilution water for 1 s before sample introduction. Repeat this procedure for tubes #5-7, incubate at 37°C.Putting this in correct scientific notation this would be:4. Aseptically add 990 μL of sterile water to tubes #1 and #2. 15. In this counting procedure, numbers such as 3b, 4c (Fig. If spreader covers entire plate, discard plate.
Do not count spiral plates with irregular distribution of colonies caused by dispensing errors. We want to hear from you.How do we know how many bacteria are in a liquid?10. Place solidified agar plates in polyethylene bags, close with ties or heat-sealer, and store inverted at 0-4.4°C.
That is, the final plates in the series should have between 30 and 300 colonies. Two segments of each wedge were counted on opposite sides of plate with 31 and 30 colonies, respectively.
If spreader covers half of plate area, count only those colonies that are well distributed in spreader-free areas.Compute SPLC unless restricted by detection of inhibitory substances in sample, excessive spreader growth, or laboratory accidents. Mix the broth culture, then aseptically add 10 μL to tube #1 and vortex.Fill in the chart below before proceeding (you do not need to turn this in).13.
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