Whole-cell Systemic Evolution of Ligands by Exponential enrichment (SELEX) is the process by which aptamers specific to target cells are developed. Aptamers selected by whole-cell SELEX have high affinity and specificity for bacterial surface molecules and live bacterial targets. To identify DNA aptamers specific to Staphylococcus aureus, we applied our rapid whole-cell SELEX method to a single-stranded ssDNA library. To improve the specificity and selectivity of the aptamers, we designed, selected, and developed two categories of aptamers that were selected by two kinds of whole-cell SELEX, by mixing and combining FACS analysis and a counter-SELEX process. Using this approach, we have developed a biosensor system that employs a high affinity aptamer for detection of target bacteria. FAM-labeled aptamer sequences with high binding to S. aureus, as determined by fluorescence spectroscopic analysis, were identified, and aptamer A14, selected by the basic whole-cell SELEX using a once-off FACS analysis, and which had a high binding affinity and specificity, was chosen. The binding assay was evaluated using FACS analysis. Our study demonstrated the development of a set of whole-cell SELEX derived aptamers specific to S. aureus; this approach can be used in the identification of other bacteria.
The objective of this study was to develop a nondestructive method to evaluate chemical components such as moisture content (MC), pH, and soluble solid content (SSC) in intact tomatoes by using hyperspectral imaging in the range of 1000–1550 nm. The mean spectra of the 95 matured tomato samples were extracted from the hyperspectral images, and multivariate calibration models were built by using partial least squares (PLS) regression with different preprocessing spectra. The results showed that the regression model developed by PLS regression based on Savitzky–Golay (S–G) first-derivative preprocessed spectra resulted in better performance for MC, pH, and the smoothing preprocessed spectra-based model resulted in better performance for SSC in intact tomatoes compared to models developed by other preprocessing methods, with correlation coefficients (rpred) of 0.81, 0.69, and 0.74 with root mean square error of prediction (RMSEP) of 0.63%, 0.06, and 0.33% Brix respectively. The full wavelengths were used to create chemical images by applying regression coefficients resulting from the best PLS regression model. These results obtained from this study clearly revealed that hyperspectral imaging, together with suitable analysis model, is a promising technology for the nondestructive prediction of chemical components in intact tomatoes.
Spectroscopy has proven to be an efficient tool for measuring the properties of meat. In this article, hyperspectral imaging (HSI) techniques are used to determine the moisture content in cooked chicken breast over the VIS/NIR (400–1,000 nm) spectral range. Moisture measurements were performed using an oven drying method. A partial least squares regression (PLSR) model was developed to extract a relationship between the HSI spectra and the moisture content. In the full wavelength range, the PLSR model possessed a maximum of 0.90 and an SEP of 0.74%. For the NIR range, the PLSR model yielded an of 0.94 and an SEP of 0.71%. The majority of the absorption peaks occurred around 760 and 970 nm, representing the water content in the samples. Finally, PLSR images were constructed to visualize the dehydration and water distribution within different sample regions. The high correlation coefficient and low prediction error from the PLSR analysis validates that HSI is an effective tool for visualizing the chemical properties of meat.
In this study, we developed a viability evaluation method for pepper (Capsicum annuum L.) seeds based on hyperspectral reflectance imaging. The reflectance spectra of pepper seeds in the 400–700 nm range are collected from hyperspectral reflectance images obtained using blue, green, and red LED illumination. A partial least squares–discriminant analysis (PLS-DA) model is developed to classify viable and non-viable seeds. Four spectral ranges generated with four types of LEDs (blue, green, red, and RGB), which were pretreated using various methods, are investigated to develop the classification models. The optimal PLS-DA model based on the standard normal variate for RGB LED illumination (400–700 nm) yields discrimination accuracies of 96.7% and 99.4% for viable seeds and nonviable seeds, respectively. The use of images based on the PLS-DA model with the first-order derivative of a 31.5-nm gap for red LED illumination (600–700 nm) yields 100% discrimination accuracy for both viable and nonviable seeds. The results indicate that a hyperspectral imaging technique based on LED light can be potentially applied to high-quality pepper seed sorting.
The increasingly common application of the near-infrared (NIR) hyperspectral imaging technique to the analysis of food powders has led to the need for optical characterization of samples. This study was aimed at exploring the feasibility of quantifying penetration depth of NIR hyperspectral imaging light for milk powder. Hyperspectral NIR reflectance images were collected for eight different milk powder products that included five brands of non-fat milk powder and three brands of whole milk powder. For each milk powder, five different powder depths ranging from 1 mm–5 mm were prepared on the top of a base layer of melamine, to test spectral-based detection of the melamine through the milk. A relationship was established between the NIR reflectance spectra (937.5–1653.7 nm) and the penetration depth was investigated by means of the partial least squares-discriminant analysis (PLS-DA) technique to classify pixels as being milk-only or a mixture of milk and melamine. With increasing milk depth, classification model accuracy was gradually decreased. The results from the 1-mm, 2-mm and 3-mm models showed that the average classification accuracy of the validation set for milk-melamine samples was reduced from 99.86% down to 94.93% as the milk depth increased from 1 mm–3 mm. As the milk depth increased to 4 mm and 5 mm, model performance deteriorated further to accuracies as low as 81.83% and 58.26%, respectively. The results suggest that a 2-mm sample depth is recommended for the screening/evaluation of milk powders using an online NIR hyperspectral imaging system similar to that used in this study.
The inspection of rice grain that may be infected by seedborne disease is important for ensuring uniform plant stands in production fields as well as preventing proliferation of some seedborne diseases. The goal of this study was to use a hyperspectral imaging (HSI) technique to find optimal wavelengths and develop a model for detecting discolored, diseased rice seed infected by bacterial panicle blight (Burkholderia glumae), a seedborne pathogen. For this purpose, the HSI data spanning the visible/near-infrared wavelength region between 400 and 1000 nm were collected for 500 sound and discolored rice seeds. For selecting optimal wavelengths to use for detecting diseased seed, a sequential forward selection (SFS) method combined with various spectral pretreatments was employed. To evaluate performance based on optimal wavelengths, support vector machine (SVM) and linear and quadratic discriminant analysis (LDA and QDA) models were developed for detection of discolored seeds. As a result, the violet and red regions of the visible spectrum were selected as key wavelengths reflecting the characteristics of the discolored rice seeds. When using only two or only three selected wavelengths, all of the classification methods achieved high classification accuracies over 90% for both the calibration and validation sample sets. The results of the study showed that only two to three wavelengths are needed to differentiate between discolored, diseased and sound rice, instead of using the entire HSI wavelength regions. This demonstrates the feasibility of developing a low cost multispectral imaging technology based on these selected wavelengths for non-destructive and high-throughput screening of diseased rice seed.