Mechanized Counting of Microbial Colonies on Rehydratable Film Media .

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Automated Counting of Microbial Colonies on Rehydratable Film Media. Mehrdad Saadat Biology 4400 Spring Semester 1999. Automated Counting of Microbial Colonies on Rehydratable Film Media. I. Background II. Methods and Results III. Conclusions. Background. Use of Petri dishes
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Mechanized Counting of Microbial Colonies on Rehydratable Film Media Mehrdad Saadat Biology 4400 Spring Semester 1999

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Automated Counting of Microbial Colonies on Rehydratable Film Media I. Background II. Methods and Results III. Conclusions

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Background Use of Petri dishes Every day, scholars use petri dishes loaded with media to culture microorganisms One use for refined is list A straightforward equation can be connected to an arrangement of cells developed on media to list the quantity of cells in the first example being tried

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Background BioCount  Developed by Apogee Systems

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Background BioCount  This is a scanner based framework utilized for distinguishing and identifying bacterial provinces in petri dishes The framework utilizes raster imaging to recognize ebbs and flows on a given media The project then acknowledges an arrangement of ebbs and flows as a settlement

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Background Previous Results The BioCount framework has ended up being dependable in numbering states on mFC agar Paula Tennant-Clegg appeared in her exploration that the BioCount framework could precisely tally coliform settlements on mFC agar when appropriately refined

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Background Rehydratable Film Media Recently the 3M organization has built up the Petrifilm ® This is a much simpler and speedier method for plating for a few reasons: no compelling reason to invest hours making a particular media no mishaps in "cutting into" the agar while streak plating

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Background Can the BioCount framework be adjusted for use with Petrifilms? Assuming this is the case, it gives a plausibility to More fast examination Greater objectivity: Elimination of inconstancy connected with manual checks Automated documentation

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Methods and Results Objectives of this Project Determine the Biocount ® parameters for ideal tallies of aggregate oxygen consuming Petrifilms Compare manual and mechanized state numbers Identify conceivable reasons for false positive and false negative mistakes in the computerized checks

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Methods and Results Procedure First: Prepare a crisp society of high-impact microorganisms Bacillus subtilis, Enterococcus fecalis, Staphylococcus aureus Once a turbid soup was acquired, it could be weakened down to give a craved convergence of bacterial cells (10 - 14 )

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Methods and Results Procedure (cont.) One ml of the weakening was then plated on the Petrifilm aseptically utilizing a pipette Proper strategy was taken after to cover the film and brood it at the temperature given by 3M™ (around 33º C) After 24 hours the movies were expelled from the hatchery At this time settlements were physically tallied

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Methods and Results Procedure (cont.) The movies were then filtered by the desktop scanner, four movies at Once plated and numbered by hand, the provinces were examined utilizing a run of the mill desktop scanner by HP at 150 dpi

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Methods and Results Hypothesis: There is not a huge contrast in the tally of bacterial settlements on a Petrifilm between a manual tally and that of one done by a robotized framework

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Methods and Results Parameters Using the parameters device, I endeavored to upgrade the parameters so that the system would distinguish a practically correct number of provinces on a given film — if this somehow managed to work I would then test the same parameters on another film.

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Methods and Results Parameters (cont.) Optimal parameters ought to finish two tests: They should take into account discovery of few, if any false positives nor any false negatives. They should precisely check the present settlements

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Methods and Results Parameters (cont.) Minimum Peak Size: 6 Radius Step: 1 Vector Radius Maximum: 3 Gray Threshhold: 135 Radius Power: 1.01 RGB shading settings: 240, 169, 25 Intensity settings: 2.15, 1.25, .08

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Methods and Results We definitely realized that the system was set to use a dark scale keeping in mind the end goal to distinguish states We additionally realize that on the off chance that we indicated a given province with the PC\'s mouse, and utilized the right catch of the mouse to tap on the settlement, we would set the dim scale parameters to that settlement\'s hues

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A territory of the film might be augmented to give more unmistakable settlements When expanded the provinces were not all the same shading and there was an angle of hues inside a given state. This seemed, by all accounts, to be a noteworthy wellspring of mistake in advancing the framework

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Methods and Results Manual Automated Film # Count Count Difference - - - - 1 745 707 38 2 286 323 17 3 525 509 16 4 498 475 23 5 442 504 62 6 387 433 46 7 240 332 92 8 413 500 87

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Methods and Results Statistical Analysis: t-test t = 1.582 At a 95% certainty level, the deviation between the computerized and manual check is not critical Therefore, the information bolster the speculation

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Conclusions Overall, there is great assention amongst manual and robotized province depends on aggregate high-impact Petrifilms If we were to pick a dull settlement as our premise for a dim scale, the project would identify anything underneath this level of obscurity If we were to pick a light state the system would recognize, light states, and dim provinces, yet it would likewise distinguish numerous false positives and foundation obstruction.

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Conclusions Areas for future work Microbiology research: Find approaches to advance province development with more noteworthy consistency of shading Computer research: Enhance the settlement discovery technique to handle shading slopes inside states

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