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Broad and overlapped FTIR absorption spectra existing between 3600 and 3000 cm−1 were resolved and improved by their deconvolution from a background scattering using a Gaussian function curve-fitting analysis with an R 2 > 0.99.
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The cells were analyzed on forward-angle light scatter and 90° light scatter using a lymphocyte gate.
The scattering curves collected from the protein samples were corrected for background scattering using intensity data collected from the reference buffer.
For determining optical properties in the presence of absorption, the absorption effects are separated out using a background scattering model with a power-law scattering dependence on wavelength [ 9, 11, 13], so that Eq. 1 can be applied.
In order to quantify the SAXS data, the background scattering was subtracted using a linear baseline, and the diffraction peaks were fitted with a Lorentzian function.
Anand et al. reported that the fluorescence of Intralipid with lipid concentration bellow 0.25% v/ v is significant between 390 and 420 nm when they attempted to use Intralipid as the background scattering to measure the time-resolved fluorescent spectrum of tyrosine dye (emission peak at 300 nm) [ 15].
Background was determined using a regression-based background mapping method.
The background scattering model used in this first step of individual analysis of each SFR spectrum is based on a 4th order polynomial, i.e. μ ′ s = a 1 (λ λ 0 ) − 1 + a 2 (λ λ 0 ) − 2 + a 3 (λ λ 0 ) − 3 + a 4 (λ λ 0 ) − 4 (10)This model allows sufficient degrees of freedom to correct for the physically incorrect assumption that [ p 1 p 2 p 3 C PF] are independent of γ.
By specification of a background scattering model within the sampled tissue (e.g. Mie and or Rayleigh scattering) it is possible to determine μ′ s and γ across the a range of wavelengths.
Buffer alone was used to correct for Raman and background scattering.
Draw the background using a pen.
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