Continuous Live-Cell Culture Imaging and Single-Cell Tracking by Computational Lensfree LED Microscopy.

Continuous cell culture monitoring as a way of investigating growth, proliferation, and kinetics of biological experiments is in high demand. However, commercially available solutions are typically expensive and large in size. Digital inline-holographic microscopes (DIHM) can provide a cost-effective alternative to conventional microscopes, bridging the gap towards live-cell culture imaging. In this work, a DIHM is built from inexpensive components and applied to different cell cultures. The images are reconstructed by computational methods and the data are analyzed with particle detection and tracking methods. Counting of cells as well as movement tracking of living cells is demonstrated, showing the feasibility of using a field-portable DIHM for basic cell culture investigation and bringing about the potential to deeply understand cell motility.

Rapid analysis of meat floss origin using a supervised machine learning-based electronic nose towards food authentication.

Authentication of meat floss origin has been highly critical for its consumers due to existing potential risks of having allergic diseases or religion perspective related to pork-containing foods. Herein, we developed and assessed a compact portable electronic nose (e-nose) comprising gas sensor array and supervised machine learning with a window time slicing method to sniff and to classify different meat floss products. We evaluated four different supervised learning methods for data classification (i.e., linear discriminant analysis (LDA), quadratic discriminant analysis (QDA), k-nearest neighbors (k-NN), and random forest (RF)). Among them, an LDA model equipped with five-window-extracted feature yielded the highest accuracy values of >99% for both validation and testing data in discriminating beef, chicken, and pork flosses. The obtained e-nose results were correlated and confirmed with the spectral data from Fourier-transform infrared (FTIR) spectroscopy and gas chromatography-mass spectrometry (GC-MS) measurements. We found that beef and chicken had similar compound groups (i.e., hydrocarbons and alcohol). Meanwhile, aldehyde compounds (e.g., dodecanal and 9-octadecanal) were found to be dominant in pork products. Based on its performance evaluation, the developed e-nose system shows promising results in food authenticity testing, which paves the way for ubiquitously detecting deception and food fraud attempts.

Intracellular gold nanoparticles influence light scattering and facilitate amplified spontaneous emission generation.

Generation of amplified stimulated emission inside mammalian cells has paved the way for a novel bioimaging and cell sensing approach. Single cells carrying gain media (e.g., fluorescent molecules) are placed inside an optical cavity, allowing the production of intracellular laser emission upon sufficient optical pumping. Here, we investigate the possibility to trigger another amplified emission phenomenon (i.e., amplified spontaneous emission or ASE) inside two different cell types, namely macrophage and epithelial cells from different species and tissues, in the presence of a poorly reflecting cavity. Furthermore, the resulting ASE properties can be enhanced by introducing plasmonic nanoparticles. The presence of gold nanoparticles (AuNPs) in rhodamine 6G-labeled A549 epithelial cells results in higher intensity and lowered ASE threshold in comparison to cells without nanoparticles, due to the effect of plasmonic field enhancement. An increase in intracellular concentration of AuNPs in rhodamine 6G-labeled macrophages is, however, responsible for the twofold increase in the ASE threshold and a reduction in the ASE intensity, dominantly due to a suppressed in and out-coupling of light at high nanoparticle concentrations.

Hybrid learning method based on feature clustering and scoring for enhanced COVID-19 breath analysis by an electronic nose.

Breath pattern analysis based on an electronic nose (e-nose), which is a noninvasive, fast, and low-cost method, has been continuously used for detecting human diseases, including the coronavirus disease 2019 (COVID-19). Nevertheless, having big data with several available features is not always beneficial because only a few of them will be relevant and useful to distinguish different breath samples (i.e., positive and negative COVID-19 samples). In this study, we develop a hybrid machine learning-based algorithm combining hierarchical agglomerative clustering analysis and permutation feature importance method to improve the data analysis of a portable e-nose for COVID-19 detection (GeNose C19). Utilizing this learning approach, we can obtain an effective and optimum feature combination, enabling the reduction by half of the number of employed sensors without downgrading the classification model performance. Based on the cross-validation test results on the training data, the hybrid algorithm can result in accuracy, sensitivity, and specificity values of (86 ± 3)%, (88 ± 6)%, and (84 ± 6)%, respectively. Meanwhile, for the testing data, a value of 87% is obtained for all the three metrics. These results exhibit the feasibility of using this hybrid filter-wrapper feature-selection method to pave the way for optimizing the GeNose C19 performance.

Nonmechanical parfocal and autofocus features based on wave propagation distribution in lensfree holographic microscopy.

Performing long-term cell observations is a non-trivial task for conventional optical microscopy, since it is usually not compatible with environments of an incubator and its temperature and humidity requirements. Lensless holographic microscopy, being entirely based on semiconductor chips without lenses and without any moving parts, has proven to be a very interesting alternative to conventional microscopy. Here, we report on the integration of a computational parfocal feature, which operates based on wave propagation distribution analysis, to perform a fast autofocusing process. This unique non-mechanical focusing approach was implemented to keep the imaged object staying in-focus during continuous long-term and real-time recordings. A light-emitting diode (LED) combined with pinhole setup was used to realize a point light source, leading to a resolution down to 2.76 µm. Our approach delivers not only in-focus sharp images of dynamic cells, but also three-dimensional (3D) information on their (x, y, z)-positions. System reliability tests were conducted inside a sealed incubator to monitor cultures of three different biological living cells (i.e., MIN6, neuroblastoma (SH-SY5Y), and Prorocentrum minimum). Altogether, this autofocusing framework enables new opportunities for highly integrated microscopic imaging and dynamic tracking of moving objects in harsh environments with large sample areas.

Intelligent Mobile Electronic Nose System Comprising a Hybrid Polymer-Functionalized Quartz Crystal Microbalance Sensor Array.

We devised a low-cost mobile electronic nose (e-nose) system using a quartz crystal microbalance (QCM) sensor array functionalized with various polymer-based thin active films (i.e., polyacrylonitrile, poly(vinylidene fluoride), poly(vinyl pyrrolidone), and poly(vinyl acetate)). It works based on the gravimetric detection principle, where the additional mass of the adsorbed molecules on the polymer surface can induce QCM resonance frequency shifts. To collect and process the obtained sensing data sets, a multichannel data acquisition (DAQ) circuitry was developed and calibrated using a function generator resulting in a device frequency resolution of 0.5 Hz. Four prepared QCM sensors demonstrated various sensitivity levels with high reproducibility and consistency under exposure to seven different volatile organic compounds (VOCs). Moreover, two types of machine learning algorithms (i.e., linear discriminant analysis and support vector machine models) were employed to differentiate and classify those tested analytes, in which classification accuracies of up to 98 and 99% could be obtained, respectively. This high-performance e-nose system is expected to be used as a versatile sensing platform for performing reliable qualitative and quantitative analyses in complex gaseous mixtures containing numerous VOCs for early disease diagnosis and environmental quality monitoring.