Water is the most common and essential active substance that supports biological reactions in living systems. For example, the conformation of macromolecules, such as proteins, is supported by the hydration water surrounding them for its proper functioning. Due to the difficulty in selectively probing the picosecond dynamics of water, the in-vitro intracellular hydration state has been left unsolved. We demonstrate terahertz (THz) spectroscopy that is sensitive to small changes in water dynamics to unravel the intracellular hydration state in the intact cells. In this study, the ordinary THz spectroscopy and living cell measurement methods are introduced, and new bio-applications based on the findings are discussed.
Rice bran layer residue evaluation commonly investigated using invasive methods by using dye. The development of a non-invasive method to evaluate the bran layer residue will enable the implementation of the methods in the rice sorting machine. Characterization of the optical properties of the rice kernel is necessary to find the effective machine vision method and setup. In this research, a preliminary experiment to develop a suitable imaging method to detect bran layer residue has been done. The fluorescence EEM of different layers of rice kernel has been measured and compared. Fluorescence imaging using a single excitation wavelength at 280 nm and UV camera has been developed to detect the bran layer residue. The results show this method has the potential to be implemented for bran residue detection and milling degree prediction.
Terahertz is a relatively new, and yet to be explored that how it interacts with many biological objects like avian eggshell. Eggshell protects internal contents of egg from contaminants and ultraviolet radiation. It also acts as a major mineral source for embryonic development. The refractive index indicating eggshell compactness could be a potential quality criterion of shell for both table and hatching egg. But a little is known about refractive index of eggshell. However, the available information is only based on destructive method which is not suitable to use in many cases due to loss of eggshell integrity. Therefore, we aim to develop a non-destructive protocol of eggshell refractive index measurement using terahertz timedomain spectroscopy (THz-TDS) which can directly derive refractive index in the terahertz region from the temporal pulses. In order to get rid of intense terahertz absorption by egg albumen, we used THz-TDS to measure reflectance of intact eggshell in the 0.2-1.2 THz. THz wave`s frequency domain interferences signal contains the information of both thickness and refractive index of the eggshell. The results revealed that this proposed method can be used in estimation of refractive index of intact eggshell with co-efficient of determination (R2) of 0.49. This non-destructive protocol could not only be used in industrial grading of eggs but also in many applied research fields such as applied optics, ornithology, evolutionary biology in the future.
Spectra of complex dielectric constant Terahertz (THz) and sub-THz range, corresponding to picosecond dynamics of the hydrogen bond network, gives us information of water molecule dynamics: rotation mode and vibration of bulk water. HeLa cell cultured on a silicon prism was measured and then analyzed by two reflection interface model and Debye- Lorentz function. Finally fraction of hydration and dynamics of bulk water molecules were estimated.
Near field sub-THz array sensor in CMOS circuit for cell detection was developed as a tool to collect quantitative cellular property. More than 1400 number of LC oscillators designed at 60 GHz are arranged in 2 dimensions periodically, and resonance frequency of each oscillator shifts by dielectric target. In simulation and experiment, frequency shifts of the CMOS sensor agreed well with dielectric constant change of water and solution on the surface. Growth of bacterial colonies was monitored successfully. Moreover, significant difference of frequency shifts was demonstrated among NHDF and HeLa, normal and cancer human cell respectively, and liquid medium.
A non-destructive method of estimating the freshness of fish is required for appropriate price setting and food safety. In particular, for determining the possibility of eating raw fish (sashimi), freshness estimation is critical. We studied such an estimation method by capturing images of fish eyes and performing image processing using the temporal changes of the luminance of pupil and iris. To detect subtle non-visible changes of these features, we used UV (375 nm) light illumination in addition to visible white light illumination. Polarization and two-channel LED techniques were used to remove strong specular reflection from the cornea of the eye and from clear-plastic wrap used to cover the fish to maintain humidity. Pupil and iris regions were automatically detected separately by image processing after the specular reflection removal process, and two types of eye contrast were defined as the ratio of mean and median pixel values of each region. Experiments using 16 Japanese dace (Tribolodon hakonensis) at 23℃ and 85% humidity for 24 hours were performed. The eye contrast of raw fish increase non-linearly in the initial period and then decreased; however, that of frozen-thawed fish decreased linearly throughout 24 hours, regardless of the lighting. Interestingly, the eye contrast using UV light showed a higher correlation with time than that using white light only in the case of raw fish within the early 6- hour period postmortem. These results show the possibility of estimating fish freshness in the initial stage when fish are eaten raw using white and UV lightings.
Defective unshu oranges (Citrus reticulate Blanco var. unshu) were sorted based on fluorescent imaging technique in a
commercial packinghouse but fresh appearing unshu were rejected due to fluorescence appearing on their peel. We
studied the various visible patterns based on colour, fluorescence and microscopic images, where even areas of the peel
that are not obviously damaged can have fluorescence, to provide a categorization of fluorescence reasons. The
categorization corresponded to: 1) hole and flow; 2) influenced by damaged or rotten fruits that have released peel oil
onto it; 3) immature or poor peel quality; 4) whitish fluorescence due to agro-chemicals and 5) variation of the growing
season. The identification of such patterns of fluorescence might be useful for citrus grading industry to take some
initiatives to make the entire automated system more efficient.
Research was conducted to acquire knowledge of the ultraviolet and visible spectrums from 300 -800 nm of some
common varieties of Japanese citrus, to investigate the best wave-lengths for fluorescence excitation and the resulting
fluorescence wave-lengths and to provide a scientific background for the best quality fluorescent imaging technique for
detecting surface defects of citrus. A Hitachi U-4000 PC-based microprocessor controlled spectrophotometer was used to
measure the absorption spectrum and a Hitachi F-4500 spectrophotometer was used for the fluorescence and excitation
spectrums. We analyzed the spectrums and the selected varieties of citrus were categorized into four groups of known
fluorescence level, namely strong, medium, weak and no fluorescence.The level of fluorescence of each variety was also
examined by using machine vision system. We found that around 340-380 nm LEDs or UV lamps are appropriate as
lighting devices for acquiring the best quality fluorescent image of the citrus varieties to examine their fluorescence
intensity. Therefore an image acquisition device was constructed with three different lighting panels with UV LED at
peak 365 nm, Blacklight blue lamps (BLB) peak at 350 nm and UV-B lamps at peak 306 nm. The results from
fluorescent images also revealed that the findings of the measured spectrums worked properly and can be used for
practical applications such as for detecting rotten, injured or damaged parts of a wide variety of citrus.
The objective of our research was to use ATR-THz spectroscopy together with chemometric for quantitative study in
food analysis. Glucose, fructose and sucrose are main component of sugar both in fresh and processed fruits. The use of
spectroscopic-based method for sugar determination is well reported especially using visible, near infrared (NIR) and
middle infrared (MIR) spectroscopy. However, the use of terahertz spectroscopy for sugar determination in fruits has
not yet been reported. In this work, a quantitative study for sugars determination using attenuated total reflectance
terahertz (ATR-THz) spectroscopy was conducted. Each samples of glucose, fructose and sucrose solution with
different concentrations were prepared respectively and their absorbance spectra between wavenumber 20 and 450 cm-1 (between 0.6 THz and 13.5 THz) were acquired using a terahertz-based Fourier Transform spectrometer (FARIS-1S,
JASCO Co., Japan). This spectrometer was equipped with a high pressure of mercury lamp as light source and a
pyroelectric sensor made from deuterated L-alanine triglycine sulfate (DLTGS) as detector. Each spectrum was
acquired using 16 cm-1 of resolution and 200 scans for averaging. The spectra of water and sugar solutions were
compared and discussed. The results showed that increasing sugar concentration caused decreasing absorbance. The
correlation between sugar concentration and its spectra was investigated using multivariate analysis. Calibration models
for glucose, fructose and sucrose determination were developed using partial least squares (PLS) regression. The
calibration model was evaluated using some parameters such as coefficient of determination (R2), standard error of
calibration (SEC), standard error of prediction (SEP), bias between actual and predicted sugar concentration value and
ratio prediction to deviation (RPD) parameter. The cross validation method was used to validate each calibration model.
It is showed that the use of ATR-THz spectroscopy combined with appropriate chemometric can be a potential for a
rapid determination of sugar concentrations.
A label-free bioaffinity sensor working in terahertz (THz) region with a nitrocellulose membrane filter was demonstrated,
which is based on the resonant transmission phenomenon and the dip in the spectra of the metal mesh device. By using
this sensor, we succeeded in the highly sensitive detection of small amounts of protein avidin-biotin complex. A distinct
change of transmittance caused by shift of the transmission dip was observed for 8 ng/mm2 (74 fmol) of horseradish
peroxidase (HRP) labeled avidin. The sensing method has broad utility for many reactions on the membrane filter as a
simple and rapid sensor.
Recent progress in the field of terahertz (THz) imaging is overviewed. First, various THz-wave sources developed and recently improved in our group are described. Second, imaging of samples can be achieved in different modes, of which we discuss here the transmission mode and the reflection-scattering mode. An emphasis in placed on the latter, which can be used to detect and determine for example the distribution of powders inside THz-transparent containers and packages. One-frequency or wide-spectrum imaging can be extended to chemical imaging, a technique by which images acquired at different THz frequencies can be combined to allow the identification of the chemical composition of the target at each spatial position. Other THz imaging applications are also discussed.
We have studied the generation of terahertz (THz) waves by optical parametric processes based on laser light scattering from the polariton mode of nonlinear crystals. Using parametric oscillation of LiNbO3 or MgO-doped LiNbO3 crystal pumped by a nano-second Q-switched Nd:YAG laser, we have realized a widely tunable coherent THz-wave sources with a simple configuration. We report the detailed characteristics of the oscillation and the radiation including tunability, spatial and temporal coherency, uni directivity, and efficiency. A Fourier transform limited THz-wave spectrum narrowing was achieved by introducing the injection seeding method. Further, we have developed a spectroscopic THz imaging system using a TPO, which allows detection and identification of drugs concealed in envelopes, by introducing the component spatial pattern analysis. Several images of the envelope are recorded at different THz frequencies and then processed. The final result is an image that reveals what substances are present in the envelope, in what quantity, and how they are distributed across the envelope area. The example presented here shows the identification of three drugs, two of which illegal, while one is an over-the-counter drug.
We have developed a novel basic technology for terahertz (THz) imaging, which allows detection and identification of chemicals by
introducing the component spatial pattern analysis. The spatial distributions of the chemicals were obtained from terahertz
multispectral transillumination images, using absorption spectra previously measured with a widely tunable THz-wave parametric
oscillator. Further we have applied this technique to the detection and identification of illicit drugs concealed in envelopes. The
samples we used were methamphetamine and MDMA, two of the most widely consumed illegal drugs in Japan, and aspirin as a reference.
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