The Most Suitable Number of Colonies on Plates for Counting

Tomasiewicz, Diane M.; Hotchkiss, D. K.; Reinbold, G. W.; Read, R. B.; Hartman, Paul A.

doi:10.4315/0362-028x-43.4.282

Cited by 76 publications

(41 citation statements)

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“…Determining CFU from Petri dishes with high throughput is an exhaustive task (7,12,13), and may be largely relieved using analog CFU inference, which infers CFU based on the time it takes a transformed culture to reach the predetermined OD of 0.1 when compared with a reference standard curve (Figure 3). For example, a culture starting from 10 3 transformed cells (CFU = 10 3 ) will reach an OD of 0.1 exactly one doubling time slower than a culture starting from 2 10 3 cells (CFU = 2 10 3 ) and so on.…”

Section: Resultsmentioning

confidence: 99%

Computer-Aided High-Throughput Cloning of Bacteria in Liquid Medium

Yehezkel

Nagar²,

Mackrants³

et al. 2011

BioTechniques

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Bacterial cloning was first introduced over a century ago and has since become one of the most useful procedures in biological research, perhaps paralleled in its ubiquity only by PCR and DNA sequencing. However, unlike PCR and sequencing, cloning has generally remained a manual, labor-intensive, low-throughput procedure. Here we address this issue by developing an automated, computer-aided bacterial cloning method using liquid medium that is based on the principles of (i) limiting dilution of bacteria, (ii) inference of colony forming units (CFUs) based on optical density (OD) readings, and (iii) verification of monoclonality using a mixture of differently colored fluorescently labeled bacteria for transformation. We demonstrate the high-throughput utility of this method by employing it as a cloning platform for a DNA synthesis process.

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Section: Resultsmentioning

confidence: 99%

Computer-Aided High-Throughput Cloning of Bacteria in Liquid Medium

Yehezkel

Nagar²,

Mackrants³

et al. 2011

BioTechniques

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“…Each sample was sonicated for 5 seconds three times using a needle ultrasonicator. The solution was then serial diluted and placed on TSB plates and the colony forming units (CFU) counted after 8 and 24 hours (Tomasiewicz, 1980). Data was represented as CFU per area (colony number on plate × dilution factor × 1.5 mL total volume / (0.01 mL sample volume × gross surface area of the glass cover plate 0.785 cm 2 ).…”

Section: Methodsmentioning

confidence: 99%

Chitosan-based nanofibrous membranes for antibacterial filter applications

Cooper

Oldinski

et al. 2013

Carbohydrate Polymers

175

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Nanofibrous membranes have drawn considerable interest for filtration applications due to their ability to withstand high fluid flux while removing micro- and nano-sized particulates from solution. The desire to introduce an antibacterial function into water filter applications presents a challenge to widespread application of fibrous membranes because the addition of chemicals or biocides may produce harmful byproducts downstream. Here, we report the development of chitosan-polycaprolactone (PCL) nanofibrous membranes to utilize the natural antibacterial property of chitosan for antibacterial water filtration. Chitosan-PCL fibers with diameters of 200–400 nm and chitosan contents of 25, 50 and 75 wt% were prepared by electrospinning. In a series of bacterial challenge tests, chitosan-PCL fibrous membranes significantly reduced Staphylococcus aureus adhesion compared to PCL fibrous membranes. In water permeability and particulate size removal tests, fibrous membranes with 25% chitosan supported the greatest water flux (~7000 L/hr/m2) with 100% removal of 300-nm particulates, while maintaining the membrane integrity. This study demonstrates the potential of chitosan-PCL nanofibrous membranes as pre-filters for water filtration systems that demonstrate combinatorial filtration and intrinsic antibacterial advantages.

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“…For manual counting, in general, the ranges in common acceptance for countable numbers of colonies on a 100 mm Petri dish are between 30 and 300 (Breed and Dotterrer. 1916;Tomasiewicz and Peeler 1980). According to this, we select 10 images with colony numbers in this range and then divide them into three groups, including Group I: 30-120 (2 images), Group II: 121-210 colonies (2 images), and Group III: 211-300 colonies (6 images).…”

Section: Colony Detectionmentioning

confidence: 99%

An automated bacterial colony counting and classification system

Chen

Zhang

2009

Inf Syst Front

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Bacterial colony enumeration is an essential tool for many widely used biomedical assays. However, bacterial colony enumerating is a low throughput, time consuming and labor intensive process since there may exist hundreds or thousands of colonies on a Petri dish, and the counting process is usually manually performed by well-trained technicians. In this paper, we introduce a fully automatic yet cost-effective bacterial colony counter which can not only count but also classify colonies. Our proposed method can recognize chromatic and achromatic images and thus can deal with both color and clear medium. In addition, the proposed method is software-centered and can accept general digital camera images as its input. The counting process includes detecting dish/plate regions, identifying colonies, separating aggregated colonies, and reporting colony counts. In order to differentiate colonies of different species, the proposed counter adopts one-class Support Vector Machine (SVM) with Radial Basis Function (RBF) as the classifier. Our proposed counter demonstrates a promising performance in terms of both precision and recall, and is robust and efficient in terms of labor-and time-savings.

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The Most Suitable Number of Colonies on Plates for Counting

Cited by 76 publications

References 0 publications

Computer-Aided High-Throughput Cloning of Bacteria in Liquid Medium

Computer-Aided High-Throughput Cloning of Bacteria in Liquid Medium

Chitosan-based nanofibrous membranes for antibacterial filter applications

An automated bacterial colony counting and classification system

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