Widely tunable fiber optical parametric oscillator synchronized with a Ti:sapphire laser for stimulated Raman scattering microscopy.

Stimulated Raman scattering (SRS) microscopy is a powerful vibrational imaging technique with high chemical specificity. However, the insufficient tuning range or speed of light sources limits the spectral range of SRS imaging and, hence, the ability to identify molecular species. Here, we present a widely tunable fiber optical parametric oscillator with a tuning range of 1470 cm-1, which can be synchronized with a Ti:sapphire laser. By using the synchronized light sources, we develop an SRS imaging system that covers the fingerprint and C-H stretching regions, without balanced detection. We validate its broadband imaging capability by visualizing a mixed polymer sample in multiple vibrational modes. We also demonstrate SRS imaging of HeLa cells, showing the applicability of our SRS microscope to biological samples.

Super-resolution vibrational imaging based on photoswitchable Raman probe.

Super-resolution vibrational microscopy is promising to increase the degree of multiplexing of nanometer-scale biological imaging because of the narrower spectral linewidth of molecular vibration compared to fluorescence. However, current techniques of super-resolution vibrational microscopy suffer from various limitations including the need for cell fixation, high power loading, or complicated detection schemes. Here, we present reversible saturable optical Raman transitions (RESORT) microscopy, which overcomes these limitations by using photoswitchable stimulated Raman scattering (SRS). We first describe a bright photoswitchable Raman probe (DAE620) and validate its signal activation and depletion characteristics when exposed to low-power (microwatt level) continuous-wave laser light. By harnessing the SRS signal depletion of DAE620 through a donut-shaped beam, we demonstrate super-resolution vibrational imaging of mammalian cells with excellent chemical specificity and spatial resolution beyond the optical diffraction limit. Our results indicate RESORT microscopy to be an effective tool with high potential for multiplexed super-resolution imaging of live cells.

Complete characterization of picosecond optical pulses by the offset frequency intensity modulation.

We present a method for characterizing the intensity waveform, spectrum, frequency chirp, and spectral phase of picosecond pulses at a moderate repetition rate of ∼100 MHz. The proposed method exploits the intensity modulation at ∼10 GHz, which is slightly offset from the integer multiple of the repetition rate of the pulses. The modulated pulses are split into two, and one is measured by an optical spectrum analyzer, whose output is detected by a lock-in amplifier, while the other is directly detected by a photodiode and its output is used as a reference signal of the lock-in amplifier. In the experiment, we demonstrate the measurement of picosecond Ti:sapphire laser pulses to investigate frequency chirp induced by self-phase modulation. We anticipate that the proposed method will be useful for the characterization of various types of picosecond pulses.

Stimulated Raman scattering spectroscopy with quantum-enhanced balanced detection.

Quantum-enhanced stimulated Raman scattering (QE-SRS) is a promising technique for highly sensitive molecular vibrational imaging and spectroscopy surpassing the shot noise limit. However, the previous demonstrations of QE-SRS utilized rather weak optical power which hinders from competing with the sensitivity of state-of-the-art SRS microscopy and spectroscopy using relatively high-power optical pulses. Here, we demonstrate SRS spectroscopy with quantum-enhanced balanced detection (QE-BD) scheme, which works even when using high-power optical pulses. We used 4-ps pulses to generate pulsed squeezed vacuum at a wavelength of 844 nm with a squeezing level of -3.28 ± 0.12 dB generated from a periodically-poled stoichiometric LiTaO3 waveguide. The squeezed vacuum was introduced to an SRS spectrometer employing a high-speed spectral scanner to acquire QE-SRS spectrum in the wavenumber range of 2000-2280 cm-1 within 50 ms. Using SRS pump pulses with an average power of 11.3 mW, we successfully obtained QE-SRS spectrum whose SNR was better than classical SRS with balanced-detection by 2.27 dB.

Phase locking of squeezed vacuum generated by a single-pass optical parametric amplifier.

In high-precision optical measurements, squeezed vacuum states are a promising resource for reducing the shot noise. To utilize a squeezed vacuum, it is important to lock the phase of the local oscillator (LO) to the squeezed light. The coherent control sideband (CCSB) scheme has been established for the precise phase locking, while the previous CCSB scheme was designed for the squeezed vacuum generated with an optical parametric oscillator (OPO). Thus the previous CCSB scheme is not applicable to squeezing by a single-pass optical parametric amplifier (OPA), which is attractive for generating broadband squeezed vacuum states. In this study, we propose a variant of CCSB scheme, which is applicable to the squeezing by single-pass OPA. In this scheme, we inject pump light and frequency-shifted signal light into an OPA crystal in the same way as the previous CCSB scheme. The parametric process in the OPA crystal generates a squeezed vacuum, amplifies the signal light, generates an idler light, and causes the pump depletion reflecting the interference of the amplified signal light and the idler light. Through the lock-in detection of the pump depletion, we can phase-lock the injected signal light to the pump light. Then, after the heterodyne detection of the signal and the idler light, we get the error signal of LO and realize the precise phase locking of LO to the squeezed quadrature. We show the feasibility of the proposed scheme by deriving the signal-to-noise ratio (SNR) of the modulated pump signal. We experimentally demonstrate the proposed scheme on pulsed squeezing by a single-pass OPA.

Probing Methionine Uptake in Live Cells by Deuterium Labeling and Stimulated Raman Scattering.

The small biomolecule methionine (Met) is a fundamental amino acid required for a vast range of biological processes such as protein synthesis, cancer metabolism, and epigenetics. However, it is still difficult to visualize the subcellular distribution of small biomolecules including Met in a minimally invasive manner. Here, we demonstrate stimulated Raman scattering (SRS) imaging of cellular uptake of deuterated methionine (d8-Met) in live HeLa cells by way of comparison to the previously used alkyne-labeled Met analogue─homopropargylglycine (Hpg). We show that the solutions of d8-Met and Hpg have similar SRS signal intensities. Furthermore, by careful image analysis with background subtraction, we succeed in the SRS imaging of cellular uptake of d8-Met with a much greater signal intensity than Hpg, possibly reflecting the increased and minimally invasive uptake kinetics of d8-Met compared with Hpg. We anticipate that d8-Met and other deuterated biomolecules will be useful for investigating metabolic processes with subcellular resolution.

Quantum-enhanced stimulated Raman scattering microscopy in a high-power regime.

Quantum-enhanced stimulated Raman scattering (QESRS) microscopy is expected to realize molecular vibrational imaging with sub-shot-noise sensitivity, so that weak signals buried in the laser shot noise can be uncovered. Nevertheless, the sensitivity of previous QESRS did not exceed that of state-of-the-art stimulated Raman scattering (SOA-SRS) microscopes mainly because of the low optical power (3 mW) of amplitude squeezed light [Nature594, 201 (2021)10.1038/s41586-021-03528-w]. Here, we present QESRS based on quantum-enhanced balanced detection (QE-BD). This method allows us to operate QESRS in a high-power regime (>30 mW) that is comparable to SOA-SRS microscopes, at the expense of 3 dB sensitivity drawback due to balanced detection. We demonstrate QESRS imaging with 2.89 dB noise reduction compared with classical balanced detection scheme. The present demonstration confirms that QESRS with QE-BD can work in the high-power regime, and paves the way for breaking the sensitivity of SOA-SRS microscopes.

Catheter-related obstruction of the right brachiocephalic vein following hemodialysis in a patient with lupus nephritis.

Vascular access is necessary for hemodialysis, and in some cases where it is difficult to establish an arteriovenous fistula or arteriovenous graft, a permanent hemodialysis catheter may be used. However, serious catheter-related complications, such as central vein stenosis or thrombosis, can occur. We herein present a case of complete brachiocephalic vein obstruction in a patient with lupus nephritis receiving hemodialysis using a tunneled hemodialysis catheter. A 64-year-old patient underwent maintenance hemodialysis while taking an anticoagulant, with a tunneled hemodialysis catheter in the right internal jugular vein, because of arteriovenous fistula failure when hemodialysis was introduced. However, the catheter was removed because of a catheter-related bloodstream infection. Following the administration of antibiotics, an arteriovenous graft was implanted between the brachial artery and axillary vein in the right arm. Surprisingly, arteriovenous graft failure and complete obstruction of the right brachiocephalic vein were observed 3 days after arteriovenous graft creation. In conclusion, we report the case of tunneled hemodialysis catheter-related complete obstruction of the right brachiocephalic vein in a lupus nephritis patient undergoing hemodialysis. Clinicians should be aware of this potential complication when tunneled hemodialysis catheters are used and consider the next vascular access type before a tunneled hemodialysis catheter has been indwelled for the long term.

Terahertz time-domain polarimetry (THz-TDP) based on the spinning E-O sampling technique: determination of precision and calibration.

We have developed a terahertz time-domain polarimetry (THz-TDP) system by applying frequency modulation to electro-optic sampling detection in a nonlinear crystal. We characterized the precision of this system in determining the polarization angles to be 1.3° for fixed time delay, and 0.5° for complete time-domain waveform. Furthermore, we calculated the Jones matrix of the optical components used for beam propagation to calibrate the induced systematic error. The advantages of employing this calibration approach are demonstrated on a sapphire crystal investigated at different sample test positions in transmission configuration, and using high resistivity Si, AlN and quartz in reflection geometry. The new THz-TDP technique has the advantage of not using any external polarizers, and therefore is not constrained by their optical performance limitations, such as restricted bandwidths and frequency-dependent extinction ratio. Finally, the THz-TDP technique can be easily implemented on existing time-domain spectroscopy (TDS) systems.

Polarization-sensitive dual-comb spectroscopy with an electro-optic modulator for determination of anisotropic optical responses of materials.

We propose and demonstrate a polarization-sensitive dual-comb spectroscopy (DCS) technique that employs an electro-optic modulator for determining the anisotropic optical responses of materials. This straightforward extension of the typical DCS setup directly provides amplitudes and phases in two mutually orthogonal directions of the electric field of light. Using this method, we determined the optic axis direction and the anisotropy in the complex refractive index of a sample whose optical parameter is well defined. We estimate a birefringence of the sample to be 5.49(55)×10-5 at a comb tooth in the 780 nm region.

Intradialytic ultrafiltration volume and vascular access outcomes: a Japan Dialysis Outcomes and Practice Patterns Study subanalysis.

INTRODUCTION: The relationship between intradialytic ultrafiltration volume and vascular access (VA) patency remains unclear. Using data from the Japan Dialysis Outcomes and Practice Patterns Study, we analyzed whether large-volume ultrafiltration was associated with VA failure in hemodialysis patients. METHODS: We included 2736 patients for whom it was possible to evaluate VA patency and bodyweight change during dialysis. Patients were divided into three groups according to the tertile of intradialytic ultrafiltration by bodyweight: low, -9.5%-3.8%; middle, 3.8%-5.1%; and high, 5.1%-13.7%. Primary VA patency was defined as the time to first VA intervention, and secondary patency as the time to creation of a new VA. Hazard ratios for VA failure were compared across groups by using Cox regression models adjusted for age, sex, body mass index, diabetes, hemoglobin and phosphorus levels, Kt/V, and erythropoiesis-stimulating agent and antiplatelet use. RESULTS: For the low, middle, and high groups, the incidences of primary and secondary VA patency were 4.7, 5.6, and 6.7 events/100 person-years and 1.3, 1.6, and 1.7 events/100 person-years, respectively. Adjusted hazard ratios for primary VA patency in the middle and high groups versus the low group were 1.16 (95% confidence interval [CI], 0.88-1.52) and 1.41 (95% CI, 1.07-1.87), respectively; those for secondary VA patency were 1.29 (95% CI, 0.78-2.13) and 1.45 (95% CI, 0.86-2.45), respectively. DISCUSSION: Large-volume ultrafiltration during dialysis tended to increase VA failure in hemodialysis patients. We thus recommend smaller ultrafiltration volumes during hemodialysis to secure VA safely.

Vascular access care and treatment practices associated with outcomes of arteriovenous fistula: international comparisons from the Dialysis Outcomes and Practice Patterns Study.

BACKGROUND: Vascular access (VA) guidelines recommend the native arteriovenous fistula (AVF) as VA of first choice for chronic hemodialysis patients. AVF management is important in hemodialysis patient care. AVF survival is associated with various physical factors, but the effects of dialysis treatment factors upon AVF survival are still not clear. METHODS: Study patients were treated at 498 dialysis facilities participating in the Dialysis Outcomes and Practice Patterns Study (DOPPS) 2 or 3 (2002-2007). Analyses included 1,183 incident hemodialysis patients (on dialysis ≤7 days and using an AVF at study entry) and 949 prevalent patients (on dialysis >7 days at DOPPS entry and using a new AVF created during study observation). AVF survival was modeled from the study entry date for incident patients and date of first AVF use for prevalent patients. Predictors of primary and final AVF survival were compared across Japan, North America and Europe/Australia/New Zealand (EUR/ANZ) with adjustments for patient characteristics. RESULTS: No meaningful relationship was seen between AVF survival and various physician and staff practices. However, patients with prior catheter use displayed higher rates of primary and final AVF failure. Final AVF failure rates were higher in facilities with higher median blood flow rates (BFR). They were also greater in North America and EUR/ANZ than in Japan, but this difference was substantially attenuated after accounting for regional differences in facility median BFR. CONCLUSION: AVF longevity differed according to the DOPPS region, and was related to prior patient catheter use and facility BFR practice. Further longitudinal studies may help demonstrate meaningful associations between VA-handling skill and patency.

A real-time terahertz time-domain polarization analyzer with 80-MHz repetition-rate femtosecond laser pulses.

We have developed a real-time terahertz time-domain polarization analyzer by using 80-MHz repetition-rate femtosecond laser pulses. Our technique is based on the spinning electro-optic sensor method, which we recently proposed and demonstrated by using a regenerative amplifier laser system; here we improve the detection scheme in order to be able to use it with a femtosecond laser oscillator with laser pulses of a much higher repetition rate. This improvement brings great advantages for realizing broadband, compact and stable real-time terahertz time-domain polarization measurement systems for scientific and industrial applications.

Measurement of 3-hydroxyisovaleric acid in urine of biotin-deficient infants and mice by HPLC.

We developed an assay for measuring urinary 3-hydroxyisovaleric acid (3-HIA) using HPLC after derivatization with 2-nitrophenylhydrazine hydrochloride (2-NPH . HCl). The derivatized 3-HIA was extracted into n-hexane and separated isocratically on a C8 reversed-phase column for fatty acids (YMC-Pack FA). We used this method to measure 3-HIA in urine extracts from mice fed a biotin-deficient diet for >4 wk and in an infant who was fed a special Japanese formula and was suspected of being biotin deficient. Urinary 3-HIA could be assayed within the range of 0.42-8.5 mmol/L with high accuracy by this method, as an indicator of biotin deficiency. Therefore, the HPLC method for 3-HIA described here may be a useful tool clinically as well as in the research laboratory.