Nonlinear spectroscopy, such as sum-frequency generation spectroscopy and coherent Raman spectroscopy, are powerful tools for analyzing transient molecular structural changes. Achieving fast and high-resolution spectroscopy with these methods requires bandwidth compression techniques to convert broadband femtosecond pulses into synchronized narrowband picosecond pulses. Here we present a novel single-pass narrowband SHG method based on a novel pulse-shaping scheme using dispersion-engineered optical filters. In our first verification experiment, we found that the SHG bandwidth after passing the filter was compressed to 1/6, and the wavelength conversion efficiency was improved by 18 times compared to the case without the filter. This result demonstrates that precision-engineered optical filters can be used as a pulse-shaping tool. This alignment-free, single-pass bandwidth compression method may be an important tool for promoting the use of nonlinear spectroscopy in a wider range of fields.
Plant disease outbreaks pose serious threats to global food security. A rigid methodology that accounts for rapid identification of the earliest point of infection caused by plant viruses is necessary. Raman spectroscopy that generates spectral signatures of cellular-level dynamics resonates the virus induced alteration in plants through moderations in spectral features. Here, we present a model study to identify the earliest point of infection. Measured spectra from healthy and virus infected Arabidopsis thaliana plants are applied to principal component analysis. We found a separation as early as 8 days post inoculation between healthy and virus infected plants.
Stimulated Raman scattering (SRS) microscopy offers label-free, high-speed measurements of small molecules in living cells and tissues, holding promise for potential biomedical applications such as drug screening and therapeutic drug monitoring. Nevertheless, with conventional SRS microscopy, fluctuations in the background Raman signal from cells and tissues limit quantitative measurements at drug concentrations below a few mM. Here we present novel SRS microscopy using temporal filtering combined with a phase-locking technique to break the detection limit due to the background Raman signals. We demonstrate that the new phase-locked system enables drug measurements at sub millimolar concentrations with linear calibration curves.
Terahertz (THz) light that resonates with the vibrational frequency of hydrated water molecules, can alter functional expression of proteins. Previous studies have shown that high-intensity THz light promotes actin fiber formation, but such mechanism is not comprehensively investigated. Here, we aim to clarify this mechanism by irradiating actin solution with THz pulsed light of lower average power and higher electric field intensity to suppress thermal effects. We compare the degree of fiber formation with and without irradiation. The result shows the contribution of electric field intensity in the accelerated actin fiber formation and clarifies the existence of non-thermal effects.
We analyze the tissue penetration of a steroid drug commonly used for the treatment of dermal and eye inflammations using phase-modulated stimulated Raman scattering (PM-SRS) microscopy. Depth-resolved imaging and penetration profile analysis reveal a clear difference in the drug penetration between the skin epithelial and the corneal epithelial tissue models due to the different barrier properties. The results suggest that PM-SRS imaging can be used to visualize the penetration pathway of topical drugs based on the spatial information of the cell and tissue structures, which should contribute to the design of drug functions.
Stimulated Raman scattering (SRS) offers high sensitivity to monitor low-concentration drugs in real time. However, the "needle" of a drug signal at millimolar concentrations is hidden in the "haystack" of tissue background Raman signals, which practically limits the specificity of the drug measurements. Here, we compare two promising solutions, namely frequency-modulated SRS (FM-SRS) and time-resolved phase-modulated SRS (PM-SRS). We show that the temporal response of FM-SRS is a simple sinusoidal function and its specificity must be compromised. However, the time-resolved PM-SRS with shaped probe pulses provides temporal high-pass filtering with steep cutoff characteristics, enabling higher specificity in the drug measurement.
In Raman-based diagnostic applications, principal component analysis (PCA) has often been used to distinguish different cell types or abnormalities. The performance of PCA greatly depends on the baseline adjustment of the measured spectra. Hence, the effect of erroneous baseline fitting on PCA requires to be addressed. Thus, we investigate the impact of baseline error for Raman spectra on PCA through the application of polynomial function with different orders in the fingerprint region (~600-1800cm-1). We found that the third order polynomial baseline fitting generated the fitted spectra closest to the mean spectrum and provided more precise PCA results.
Mastitis is a common disease in dairy cows and is considered to be one of the most intractable diseases in the world. We analyzed Raman spectra of milk samples from cows that have been treated for mastitis and have recovered, and those from cows that have no history of mastitis. The result shows that signal components derived from carotenoids and unsaturated fatty acids can be used as markers to predict mastitis history. Our new method based on simple and direct milk measurements will be a powerful tool to determine the prevalence and severity of mastitis in future field diagnosis applications.
Therapeutic drug monitoring (TDM) is required for an optimal treatment plan to control the dosage of high-risk drugs by monitoring their blood concentration. Currently, a combination of liquid chromatography and tandem mass spectrometry is used for TDM. However, this method requires expertise and skills in sample pretreatment and is not available in all hospitals. Raman spectroscopy allows us to quantify substances in biological samples with only simple pretreatment. Two methods have been reported for detecting low-concentration substances using Raman spectroscopy: surface-enhanced Raman spectroscopy and drop coating deposition Raman spectroscopy. However, it is difficult to quantify the concentration by these methods because the Raman spectra are measured in a dry state. Here, we present a new method to quantify low-concentration pharmaceutical analytes using droplet evaporation Raman spectroscopy. Methotrexate (MTX), one of the immunosuppressive drugs, is reported to cause adverse effects above 10 μM of its blood concentration 24 hours after administration. To quantify low-concentration MTX, we drop the solution onto a superhydrophobic substrate, and simultaneously measure the Raman spectra and the volume of the droplets before completely dried. In particular, we control the evaporation rate of the droplets in a humidified environment, allowing Raman measurements with sufficient exposure time. The initial concentration of the solution is determined from the measured drug concentration during evaporation and the concentration ratio obtained from the volume measurement. Using the new method, we can quantify the concentration of MTX at 50 μM, which is in the order of magnitude required for clinical use.
We propose a new method for therapeutic drug monitoring using Raman spectroscopy to quantify the concentration of analytes during droplet evaporation. The proposed method is demonstrated for quantitative measurement of methotrexate down to 0.5 mM
Quantitative measurements of a small amount of chemicals in label-free tissue imaging by conventional SRS microscopy remain challenging because of background signals. We present a time-resolved, phase-modulated (PM) SRS microscopy method with increased signal contrast. In addition to removing background signals generated via amplitude modulation, PM-SRS can reduce intrinsic tissue background signals by temporally separating the excitation and detection processes. Furthermore, polarization artifacts in tissues can also be removed by rapidly modulating the relative delay between the pump and probe pulses. This new technology enables robust quantitative measurements in tissue and extends the potential of SRS imaging in biomedical applications.
The development of a technology that allows for analyzing microscopic spatial distribution and dynamics of small gaseous molecules such as inhalational anesthetics and odors would advance our understanding of its biological activities in living cells. However, direct observation of such small molecules by optical microscopy is still challenging. We propose a new pump–probe stimulated Raman scattering (SRS) microscopy method for studying the localization, transport and metabolism of gaseous molecules in a living organism in a label-free manner. A technical challenge is how to detect the Raman signal of a small amount of drug molecules that is typically overwhelmed by unwanted nonlinear background, including nonresonant background and coherent Raman scattering of surrounding cells and tissues. In particular, the latter Raman-induced background is essentially inevitable in most standard coherent anti-Stokes Raman scattering (CARS) and SRS systems. We show that these background issues can be overcome by introducing a new pump–probe, time-resolved SRS detection approach coupled with a pair of spectrally-focused, asymmetrically shaped probe pulses (T. Ito et al. APL Photonics (2018)). In the pump–probe scheme, a long-lived vibration of the targeted molecules can be efficiently probed after short-lived vibrations of other background molecules such as water and fatty acids become silent. This unique lifetime-selective signal detection provides a significantly enhanced vibrational signal contrast. As a proof-of-concept experiment, we demonstrate that the passive transport of inhalational anesthetic molecules from aqueous solution to adipose cells can be monitored by time-lapse SRS imaging.
KEYWORDS: Holography, Holograms, Multiplexing, Diffraction, Optical amplifiers, Energy efficiency, Signal processing, Two wave mixing, Phase conjugation, Four wave mixing
Copying speed is an important characteristic for optical read-only memory (ROM) replication systems. The copying
speed of holographic ROM replication is, however, limited by small energy efficiency of the optical system due to the
small diffraction efficiency of multiplexed holograms. In this paper we propose new holographic ROM replication
systems with a photorefractive amplifier, and analyze the speed gain performance. We improve energy efficiency
significantly and speed up replication by amplifying weak diffraction signal beams using photorefractive wave mixing.
Our new theory and numerical calculations revealed that achievable speed gain can be evaluated from only a single
dimensionless parameter that is the product of the three as follows: (i) the pump beam intensity ratio in the amplifier, (ii)
the ratio of the photopolymer and photorefractive sensitivities, and (iii) the dynamic range per hologram of the copy
medium. In current holographic recording systems, a practical copying speed gain of more than 10 is achievable with
currently available photorefractive materials.
We propose a new optical intersatellite communications system with a phase conjugate mirror (PCM) in formation flying (FF). In conventional optical intersatellite communications, high-accurate target acquisition and tracking are required for both the transmitter and the receiver. In our system with a PCM, when a control beam from the receiver is captured by a PCM in the transmitter, the signal beam from the transmitter introduced back to the receiver as its phase-conjugate replica. Thus, it is not necessary for the transmitter to target the receiver. Another advantage of using a PCM is that we can utilize spatial filtering. Background noise by sunlight with the laser wavelength can also be efficiently suppressed by a spatial phase modulation/demodulation and filtering processes using phase compensation by the PCM, which leads to the improvement of the signal-to-noise ratio (SNR) and hence provides high data transmission rates in the system. In order to efficiently filter out the background noise, a large beam propagation angle is required in spatial filtering. We spatially modulate the background noise by the diffuser and reduce the beam diameter by the expansion/downscale optical system as a method to enlarge the beam propagation angle. In this paper, we show that our system can separate the noise from the signal by using the expansion/downscale optical system even under spatial phase modulation. In the analysis, the SNR is 32.6[dB] at scale=8.0×104, when a spatial phase modulation by the diffuser is θ=1.5×10-5[rad].
Volume holographic recording is a promising solution for next- generation optical disc storage that has a high capacity more than 1 TB. This huge capacity is achieved by superimposing many holograms, each of which has millions of bits, at the same recording spot. We proposed a new technique, Spatial Spread Spectrum (SSS) multiplex recording. Unlike conventional multiplex holography based on Bragg effect of thick holograms, our technique utilizes spatial phase modulation and demodulation of the signal beam itself with a random diffuser to address the multiplexed page data. SSS multiplexing is additionally combined with other multiplexing methods, and provides further improvement of the total capacity of holographic storage. In this paper we experimentally verify the basic recording and readout feasibility, and investigate the shift selectivity and the aligning margin of the SSS holographic recording that are an important factor to determine the tolerance against vibration. It is shown that a clear 2-dimensional image is successfully reconstructed from the hologram even in the case the central part of the diffused signal beam is blocked in recording, and that a sharp shift selectivity about 5 microns was obtained by a diffuser with a diffusion angle of 15 degree, and the aligning margin for a sufficient SNR was approximately 1 micron.
The purpose of this study is to apply a free-space optical interconnection to a reconfigurable board-to-board connection where the wiring patterns connecting boards are optically formed without electrical-optical conversion. We regard a photorefractive bi-directional connection module (PBCM) based on a mutually pumped phase conjugate mirror as a key device to construct such a connection network and employ PBCMs at input/output interfaces of each board. Although optical behaviors of PBCM are influenced by the exposure conditions, we especially focus on the diameter of beams illuminating photorefractive media placed inside PBCM so as to find some geometrical restrictions in a design of networking system. Through numerical analyses, we show a sample configuration of PBCM for the board-to-board interconnection and present a conceptual design of input/output interface.
We propose a fault-tolerant holographic memory (FTHM) composed of a pair of photorefractive crystals. This memory offers not only non-destructive readout but also data restoring function by only pure optical operations without any electrical controls. In writing process, the same holographic data are simultaneously recorded as index gratings to the crystals laid out in series. In reading process, a reading beam is diffracted by the index gratings in each crystal. Here, some of the diffraction beams are detected as an output beam, and the others are used as a feedback beam. The hologram in each crystal is continuously refreshed by the feedback beam from the other crystal since the feedback beam has the same information as the original holographic data. When the data refreshing effect by the feedback beams sufficiently exceeds the erasure effect by the exposure of the reading beam, the stored data are always maintained. Furthermore, even if a certain fault such as vibration and stray beam incidence happens, the lost data in one crystal are all-optically restored as long as the corresponding holographic data remain in the other crystal. The experiment with a two-dimensional image is carried out for the purpose of checking the data restoring function in FTHM. The two-dimensional image divided in quarters is recorded as into a pair of 45°-cut BaTiO3 crystals, and the original holographic data is successfully restored by the refreshing effect in the case that a quarter of the image in the one crystal is partially lost.
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