![]() ; Lin, Jake ![]() in BioTechniques (2013) Microorganisms often form multicellular structures,such as biofilms and structured colonies, which can influence the organism’s virulence, drug resistance, and adherence to medical devices. Phenotypic ... [more ▼] Microorganisms often form multicellular structures,such as biofilms and structured colonies, which can influence the organism’s virulence, drug resistance, and adherence to medical devices. Phenotypic classification of these structures has traditionally relied on qualitative scoring systems that limit detailed phenotypic comparisons between strains. Automated imaging and quantitative analysis have the potential to improve the speed and accuracy of experiments designed to study the genetic and molecular networks underlying different morphological traits. We have developeda platform that uses automated image analysis and pattern recognition to quantify phenotypic signatures of yeast colonies. The strategy enables quantitative analysis of individual colonies, measured at a single time point or over a series of time-lapse images, as well as the classification of distinct colony shapes based on image-derived features. Phenotypic changes in colonymorphology can be expressed achanges in feature space trajectories over time, thereby enabling the visualization and quantitative analysis of morphological development. To facilitate data exploration, results are plotted dynamically through an interactive web application that integrates the raw and processed images across all time points, allowing exploration of the image-based features and principal components associated with morphological development. The web application YIMAA is available at http://yimaa.cs.tut.fi. [less ▲] Detailed reference viewed: 153 (3 UL)![]() ![]() ; Friederich, Evelyne ![]() in BioTechniques (1999), 26(3), 484-6488490 The concentration of proteins in cells is an important parameter that determines how a protein will interact with other proteins or pharmacological agents. Recent developments in Western blotting ... [more ▼] The concentration of proteins in cells is an important parameter that determines how a protein will interact with other proteins or pharmacological agents. Recent developments in Western blotting techniques have now made this a method of choice to measure protein concentration in complex solutions such as total cell extracts. We show that detection of Cy5-coupled secondary antibodies by PhosphorImager analysis produces signals that approach linearity with respect to protein concentration over a 20-fold range. We used this technique to estimate cellular levels of zyxin, which is an important protein component of the actin cytoskeleton in mammalian cells. By producing specific protein standards based on sequences that are available from public databases, it is now possible to estimate the concentration of almost any protein by this technique. [less ▲] Detailed reference viewed: 100 (0 UL) |
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