A comparison between the commonly used absorption spectrophotometry and a more recent approach known as structured laser illumination planar imaging (SLIPI) is presented for the characterization of scattering and absorbing liquids. Water solutions of milk and coffee are, respectively, investigated for 10 different levels of turbidity. For the milk solutions, scattering is the dominant process, while the coffee solutions have a high level of absorption. Measurements of the extinction coefficient are performed at both λ=450 nm and λ=638 nm and the ratio of their values has been extracted. We show that the turbidity limit of valid transmission measurements is reached at an optical depth of OD∼2.4, corresponding here to an extinction coefficient of μe=0.60 mm-1 when using a modern absorption spectrometer having a spatial Fourier filter prior to detection. Above this value, errors are induced due to the contribution of scattered and multiply scattered photons reaching the detector. On the contrary, the SLIPI measurements were found to be very reliable, even for an extinction coefficient three times as high, where μe=1.80 mm-1. This improvement is due to the capability of the technique in efficiently suppressing the contribution from multiple light scattering.
Refractive index retrieval is possible using the transport intensity equation (TIE), which presents advantages over interferometric techniques. The TIE method is valid only for paraxial ray assumptions. However, diffraction can nullify these TIE model assumptions. Therefore, the refractive index is problematic for reconstruction in three-dimensions (3D) using a set of defocused images, as diffraction effects become prominent. We propose a method to recover the 3D refractive index by combining TIE and deconvolution. A brightfield (BF) microscope was then constructed to apply the proposed technique. A microsphere was used as a sample with well-known properties. The deconvolution of the BF-images of the sample using the microscope’s 3D point spread function led to significantly reduced diffraction effects. TIE was then applied for each set of three images. Applying TIE without taking into account diffraction failed to reconstruct the 3D refractive index. Taking diffraction into account, the refractive index of the sample was clearly recovered, and the sectioning effect of the microsphere was highlighted, leading to a determination of its size. This work is of great significance in improving the 3D reconstruction of the refractive index using the TIE method.
In this work, we have used multispectral imaging technology to classify cassava leaves infected by African mosaic virus by the use of their unique spectral finger print. The spectra are extracted from transmission, reflection and diffusion of their multispectral images; they have been then analyzed with statistical multivariate analysis techniques. Principal component analysis (PCA) has been used followed by K-means and Ascending Hierarchical Classification (AHC) to endorse the classification. The contribution of this work is the use of multispectral imagery which binds both spatial and spectral information to differentiate and sort infected leaves. The results show that the multimodal and imaging spectroscopy may allow blind identification and characterization of infected leaves.
Rice is staple in the African habitats menu. Bacterial wilt (BLB) and leaf streak (BLS) are some of the phytopathological diseases which restrain rice production around the world. In this paper, multi-spectral and multimodal imaging techniques have been developed to characterize the rice leaves with symptoms of bacterial wilt (BLB) and leaf streak (BLS), and to provide information on their effects, in order to reduce their spread. First, we recorded microscopic and spectroscopic images of the samples using multimodal and multispectral microscope, with spectral region ranging from UV to NIR, for each mode. Then, we extracted the spectral footprints of the cells constituents, in transmission, reflection and scattering from the spectral images. Applying multivariate statistical analysis methods to this optical spectra allowed us to characterize the effect of bacterial rice leaves caused by Xanthomonas oryzae strains. The results of the proposed technique can be useful for easy identification of this type of infection, and can serve as routine approach in biochemical and agronomic laboratories.
Most commercially available ground coffees are processed from Robusta or Arabica coffee beans. In this work, we report on the potential of Structured Laser Illumination Planar Imaging (SLIPI) technique for the classification of five types of Robusta and Arabica commercial ground coffee samples (Familial, Belier, Brazil, Colombia and Malaga). This classification is made, here, from the measurement of the extinction coefficient µ e and of the optical depth OD by means of SLIPI. The proposed technique offers the advantage of eliminating the light intensity from photons which have been multiply scattered in the coffee solution, leading to an accurate and reliable measurement of µ e . Data analysis uses the chemometric techniques of Principal Component Anaysis (PCA) for variable selection and Hierarchical Cluster Analysis (HCA) for classification. The chemometric model demonstrates the potential of this approach for practical assessment of coffee grades by correctly classifying the coffee samples according to their species.
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