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Older (.OPJ), but not newer (.OPJU), Origin project files can be read by the open-source LabPlot or SciDAVis software. The files can also be read by QtiPlot but only with a paid "Pro" version. Finally the liborigin[1] library can also read .OPJ files such as by using the opj2dat script, which exports the data tables contained in the file.
There is an origin file viewer to see data and charts made with origin. The actual software is Version 9.6.5. This software can convert newer OPJU files to older OPJ files for older versions of Origin. [3]
Origin was first created for use solely with microcalorimeters manufactured by MicroCal Inc. (acquired by Malvern Instruments in 2014[4]) The software was used to graph the instruments data, and perform nonlinear curve fitting and parameter calculation.
The MicroCal iTC200 is a highly sensitive, low volume isothermal titration calorimeter for the label-free in solution study of biomolecular interactions. It delivers direct measurement of all binding parameters in a single experiment and can analyze weak to high affinity binders, using as little as 10 µg sample. Semi-automated maintenance minimizes operator intervention and the system is upgradable to the fully automated MicroCal Auto-iTC200, making it ideal for laboratories where speed, sensitivity and the ability to accommodate higher workloads in the future are paramount.
A molecular definition of the mechanism conferring bacterial multidrug resistance is clinically crucial and today methods for quantitative determination of the uptake of antimicrobial agents with single cell resolution are missing. Using the naturally occurring fluorescence of antibacterial agents after deep ultraviolet (DUV) excitation, we developed a method to non-invasively monitor the quinolones uptake in single bacteria. Our approach is based on a DUV fluorescence microscope coupled to a synchrotron beamline providing tuneable excitation from 200 to 600 nm. A full spectrum was acquired at each pixel of the image, to study the DUV excited fluorescence emitted from quinolones within single bacteria. Measuring spectra allowed us to separate the antibiotic fluorescence from the autofluorescence contribution. By performing spectroscopic analysis, the quantification of the antibiotic signal was possible. To our knowledge, this is the first time that the intracellular accumulation of a clinical antibitiotic could be determined and discussed in relation with the level of drug susceptibility for a multiresistant strain. This method is especially important to follow the behavior of quinolone molecules at individual cell level, to quantify the intracellular concentration of the antibiotic and develop new strategies to combat the dissemination of MDR-bacteria. In addition, this original approach also indicates the heterogeneity of bacterial population when the same strain is under environmental stress like antibiotic attack.
Copyright: © 2012 Kaščáková et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Here we report the quantitative single-cell fluorescence microscopic study of Enterobacter aerogenes resistant strains using a new non-invasive method preserving the antibiotic structure which have allowed us to monitor the antibiotic uptake depending on the efflux pump activity. Considering the natural fluorescence of clinically used quinolone, we use synchrotron radiation DUV imaging and the synchrotron radiation DUV microspectroscopy as new methods to investigate the drug accumulation inside individual bacteria. Two fluorescence microscopes were used: for DUV imaging we used a DUV compatible full-field microscope, whereas the DUV microspectroscopy was achieved by using the microscope, which allows collecting the fluorescence spectra. Fleroxacin (Fle) was chosen as target quinolone to test the concept in vitro. Since the fluorimetric method of Fle uptake on digested bacterial masses has been previously validated [22], [23], we used this method as our internal standard. The intrabacterial concentration has been studied in two bacterial strains of Enterobacter aerogenes: a resistant isolate EA289 that overproduced the broad spectrum AcrAB-TolC efflux pump, and the tolC deficient derivative strain EA298 [24]. The activity of efflux pump on antibiotic uptake has been assessed using co-incubation with glucose (Glu) as well as carbonyl cyanide m-chlorophenyl hydrazone (CCCP). CCCP is a powerful uncoupler of the proton motive force (PMF) that collapses the membrane energy, consequently, used at low concentrations, it inhibits the drug transport through the inner membrane. Belonging to the group of efflux pump blockers/modulators, it is used for a long time to study the antibiotic expel by Gram-negative efflux pumps (for recent reviews see [25], [26]). These results have important implications for the understanding of intracellular accumulation of quinolones in single multidrug resistant clinical bacteria and to develop original ways to combat resistance mechanisms associated with membrane permeability.
We used synchrotron-radiation DUV microspectroscopy as a method to accurately quantify the antibiotic fluorescence in single bacteria. Measuring the spectra from single bacteria allowed monitoring of antibiotic fluorescence together with autofluorescence contribution. By doing spectroscopic analysis, the autofluorescence was successfully subtracted and consequently, the quantification of the signal from different conditions of co-incubations was possible.
In addition, we clearly demonstrated that the fluorescence drug signal from individual bacteria varied, within in a uniformly treated population. Considering that measurements are done on single bacteria, our technique can be further developed and applied to probing a bacterial population, and generate statistical information about individual bacterial uptake since isogenic microbial populations contain substantial cell-to-cell differences in physiological parameters such as growth rate, resistance to stress and regulatory circuit output (for a review see [29]). Regarding this point, it is important to mention that Long et al [38] recently reported that the quantification by single-cell fluorescence microscopy of bacterial signalling responses to AI-1 and AI-2 autoinducers involved in quorum sensing regulation, indicates a coherent response across the population but some cell-to-cell variations. This is especially important taking into account the bacterial adaptability to external stresses and antibiotic attacks and reflects the recent observation about the bacterial charity [28]. In addition, this result correlates with the large and rapid change involving the expression of transporters in bacterial membrane faced with external antibacterial agents (including antibiotics, biocides, etc). This efficient alteration of transporters expression rapidly modifies the membrane permeability and the associated-drug uptake [39], [40]. This original approach indicates the heterogeneity of bacterial population regarding the antibiotic accumulation and may provide appropriate clues to understand the role of ciprofloxacin in the bacterial adaptation and persister formation as recently reported by Dörr et al [41].
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