26 research outputs found

    Optimization of fixation methods for observation of bacterial cell morphology and surface ultrastructures by atomic force microscopy

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    Fixation ability of five common fixation solutions, including 2.5% glutaraldehyde, 10% formalin, 4% paraformaldehyde, methanol/acetone (1:1), and ethanol/acetic acid (3:1) were evaluated by using atomic force microscopy in the present study. Three model bacteria, i.e., Escherichia coli, Pseudomonas putida, and Bacillus subtilis were applied to observe the above fixation methods for the morphology preservation of bacterial cells and surface ultrastructures. All the fixation methods could effectively preserve cell morphology. However, for preserving bacterial surface ultrastructures, the methods applying aldehyde fixations performed much better than those using alcohols, since the alcohols could detach the surface filaments (i.e., flagella and pili) significantly. Based on the quantitative and qualitative assessments, the 2.5% glutaraldehyde was proposed as a promising fixation solution both for observing morphology of both bacterial cell and surface ultrastructures, while the methonal/acetone mixture was the worst fixation solution which may obtain unreliable results

    At least four distinct blue cationic phthalocyanine dyes sold as “alcian blue” raises the question: what is alcian blue?

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    We investigated physicochemical characteristics of dye lots sold as “alcian blue” using the Biological Stain Commission (BSC) precipitation test, differential scanning calorimetry, high performance liquid chromatography, thin layer chromatography and UV/visible spectroscopy. Four blue phthalocyanine dyes were detected in 11 commercial dye lots. These four included the original ingrain blue 1 CI 74000 dye and the dye sold with the name “alcian blue pyridine variant”; we discuss also the possible identity of the additional two dyes. A proposed extension to the BSC analytic scheme is presented that could distinguish three categories of commercial alcian blue dyes from each other and from the original alcian blue 8G
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