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For transport experiments, however, where the changes in the local macromolecule concentration with time are measured, a reference buffer intensity is not necessarily required.
A single buffer profile (IBuf) was scaled by the same constant to serve as a buffer intensity reference.
A probe was called positive (detected above background) when its signal intensity was above the buffer intensity on at least 50% of slides within a given sample type.
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Under the experimental conditions employed (100 μM meropenem and a suspension of NDM-1 E. coli cells with an optical density at 600 nm (OD600) of 2.5 in sodium phosphate buffer), the intensity of the H signals from the methyl groups is about fivefold higher than from cells alone.
The scattering images obtained were averaged and buffer scattering intensities subtracted using PRIMUS software.
For a thicker LT buffer, the PL intensity is weaker.
One hundred µl of the resulting cell extracts were mixed with 1 ml of the same buffer, and fluorescence intensity was determined using an Applied Biosystems Citofluor 4000 Fluorescence Multi-well plate reader (excitation 490 nm, emission 519 nm).
After centrifuging and rinsing in FACS buffer, FITC fluorescence intensity of cells was measured using the FACSAria.
Normalization for beamline intensity, buffer subtraction and merging of data were carried out using proc.py software developed by the beamline staff (Allaire and Yang, 2011).
All buffer blanks produced intensities equivalent to the noise floor (an intensity of 1000 in the 95th percentile on the green detection channel).
Reduced sensitivity typically acts by a buffer that lowers the intensity of natural selection on the direct character; the lower intensity of selection is what leads to a decay in performance of the direct character.
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