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.
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.
Biofilms formed on the surface of agro-food processing facilities can cause food poisoning by providing an environment in which bacteria can be cultured. Therefore, hygiene management through initial detection is important. This study aimed to assess the feasibility of detecting Escherichia coli (E. coli) and Salmonella typhimurium (S. typhimurium) on the surface of food processing facilities by using fluorescence hyperspectral imaging. E. coli and S. typhimurium were cultured on high-density polyethylene and stainless steel coupons, which are the main materials used in food processing facilities. We obtained fluorescence hyperspectral images for the range of 420–730 nm by emitting UV light from a 365 nm UV light source. The images were used to perform discriminant analyses (linear discriminant analysis, k-nearest neighbor analysis, and partial-least squares discriminant analysis) to identify and classify coupons on which bacteria could be cultured. The discriminant performances of specificity and sensitivity for E. coli (1–4 log CFU·cm−2) and S. typhimurium (1–6 log CFU·cm−2) were over 90% for most machine learning models used, and the highest performances were generally obtained from the k-nearest neighbor (k-NN) model. The application of the learning model to the hyperspectral image confirmed that the biofilm detection was well performed. This result indicates the possibility of rapidly inspecting biofilms using fluorescence hyperspectral images.
A Tungsten-Halogen (TH) lamp is the most popular light source in NIR spectroscopy and hyperspectral imaging, which requires a warm-up to reach very high temperatures of up to 250 °C and take a long time for radiation stabilization. Consequently, it has a large enough volume to enable heat dissipation to prevent the thermal runaway of the electric circuit and turn out its power efficiency very low. These are major barriers for miniaturizing spectral systems and hyperspectral imaging devices. However, TH lamps can be replaced by pc-NIR LEDs in order to avoid high temperature and large volume. We compared the spectral emission of the available commercial pc-NIR LEDs under the same condition. As a replacement for the TH lamp, the VIS + NIR LED module was developed to combine a warm-white LED and pc-NIR LEDs. In order to feature out the availability of the VIS + NIR LED module against the TH lamp, they were used as the light source for evaluating the Soluble Solid Content (SSC) of an apple through VIS-NIR spectroscopy. The results show a remarkable feasibility in the performance of the partial least square (PLS) model using the VIS + NIR LED module; during PLS calibration, the correlation coefficient (R) values are 0.664 and 0.701, and the Mean Square Error (MSE) values are 0.681 and 0.602 for the TH lamp and VIS + NIR LED module, respectively. In VIS-NIR spectroscopy, this study indicates that the TH lamp could be replaceable with a warm-white LED and pc-NIR LEDs.
This research aims to design and fabricate a system to measure the capsaicinoid content of red pepper powder in a non-destructive and rapid method using visible and near infrared spectroscopy (VNIR). The developed system scans a well-leveled powder surface continuously to minimize the influence of the placenta distribution, thus acquiring stable and representative reflectance spectra. The system incorporates flat belts driven by a sample input hopper and stepping motor, a powder surface leveler, charge-coupled device (CCD) image sensor-embedded VNIR spectrometer, fiber optic probe, and tungsten halogen lamp, and an automated reference measuring unit with a reference panel to measure the standard spectrum. The operation program includes device interface, standard reflectivity measurement, and a graphical user interface to measure the capsaicinoid content. A partial least square regression (PLSR) model was developed to predict the capsaicinoid content; 44 red pepper powder samples whose measured capsaicinoid content ranged 13.45–159.48 mg/100 g by per high-performance liquid chromatography (HPLC) and 1242 VNIR absorbance spectra acquired by the pungency measurement system were used. The determination coefficient of validation (RV2) and standard error of prediction (SEP) for the model with the first-order derivative pretreatment method for Korean red pepper powder were 0.8484 and ±13.6388 mg/100 g, respectively.