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Quantitative measurements of water content within a single cell are notoriously difficult. In this work, we introduce a single-shot optical method for tracking the intracellular water content, by mass and volume, of a single cell at video rate. We utilize quantitative phase imaging and a priori knowledge of a spherical cellular geometry, leveraging a two-component mixture model to compute the intracellular water content. We apply this technique to study CHO-K1 cells responding to a pulsed electric field, which induces membrane permeabilization and rapid water influx or efflux depending upon the osmotic environment. The effects of mercury and gadolinium on water uptake in Jurkat cells following electropermeabilization are also examined.
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Brillouin microscopy has recently emerged as a powerful tool for mechanical property measurements in biomedical sensing and imaging applications. Impulsive stimulated Brillouin scattering (ISBS) microscopy has been proposed for faster and more accurate measurements, which do not rely on stable narrow-band lasers and thermally-drifting etalon-based spectrometers. However, the spectral resolution of ISBS-based signal has not been significantly explored. In this report, the ISBS spectral profile has been investigated as a function of the pump beam's spatial geometry, and novel methodologies have been developed for accurate spectral assessment. The ISBS linewidth was found to consistently decrease with increasing pump-beam diameter. These findings provide the means for improved spectral resolution measurements and pave the way to broader applications of ISBS microscopy.
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The purpose of this explorative study is to determine if critically ill patients experience cardiac atrophy that can be quantified as a loss of left ventricular mass (LVM) and thus detected by echocardiography. DESIGN: Retrospective single-center cohort study. SETTING: Patients admitted to a tertiary medical center in Boston, MA. PATIENTS: Adult critically ill patients with ICU length of stay greater than or equal to 5 days. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We conducted a retrospective cohort study of 68 patients, of which 42 were included in the final analysis (mean age 60.9 ± 19.2 yr; 47.6% male). The median length of ICU stay was 11.3 days (interquartile range, 6.8-20.1 d). A decrease in mean LVM over the course of admission for critical illness was observed (median 189.11 g [162.82-240.20 g] vs 176.69 g [142.37-226.26 g]; p = 0.01). After adjusting for sex, age, fluid balance, ICU type, dietary orders, time between echocardiograms, and vasopressor use, this decrease in LVM remained consistent (mean difference, -21.30 g; 95% CI, -41.85 to -0.74; p = 0.04). Relative wall thickness (RWT) did not change during admission. CONCLUSIONS: These data reveal that a loss of LVM is present in patients over their ICU stay without a corresponding change in RWT, consistent with cardiac atrophy. Future prospective studies are needed to confirm these findings and identify possible sequelae of this finding.
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The electronic or molecular mechanisms that initiate photobiomodulation (PBM) in cells are not yet fully understood. The porcine complex III (C-III) of the electron transport chain was characterized with transient absorption spectroscopy (TAS). We then applied our recently developed continuous wave laser coupled TAS procedure (CW-TAS) to investigate the effect of red light irradiances on the heme dynamics of C-III in its c1 reduced state. The time constants were found to be 3.3 ± 0.3 ps for vibrational cooling of the oxidized state and 4.9 ± 0.4 ps for rebinding of the photodissociated axial ligand of the c1 reduced state. The analysis of the CW-TAS procedure yielded no significant changes in the C-III heme dynamics. We rule out the possibility of 635 nm CW light at 4.7 mW/cm2 inducing a PBM effect on the heme dynamic of C-III, specifically with the photodissociation of its axial ligand.
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Photobiomodulation (PBM) describes the use of low irradiance light in the red to near-infrared wavelength range to stimulate biological effects in tissue, and many biological and spectroscopic techniques are used to study PBM. However, these techniques focus on the products or downstream effects rather than the electronic transitions that initiate the PBM processes. This study presents a novel approach to studying low irradiance light exposures on individual proteins and/or protein complexes by combining a continuous wave (CW) laser diode with femtosecond transient absorption spectroscopy (TAS), coined here as CW-TAS, and tests the system on reduced cytochrome c (Cyt c) for proof of principle. TAS was conducted using a 532-nm excitation pump beam and a 350-600 nm supercontinuum probe. CW laser diodes with wavelengths of 450 nm, 635 nm, and 808 nm were interchangeably fiber coupled into the HELIOS Fire. Samples of Cyt c were tested by TAS using a pump power of 15 µW, both with and without CW exposure. CW exposures were carried out with irradiances of 1.60 and 3.20 mW/cm2, except for 808 nm, which was only tested at 1.60 mW/cm2. Both kinetic and global analyses were performed on the TAS data and the time constants for sets with and without CW exposures were compared. The TAS data for Cyt c with the full dosage of CW exposures did not alter the TAS data distinguishably from the control data. No new electronic transient signals were observed beyond the background when testing Cyt c with the CW exposures. Kinetic analysis confirmed that existing transients did not deviate beyond uncertainty. Global time constants for Cyt c were calculated to be 0.25 ± 0.03 ps and 5.1 ± 0.3 ps for the control study, and the time constants for the CW exposed Cyt c were not significantly different. This study concludes that CW irradiation, at doses delivered, does not alter the transient absorption data of Cyt c. The CW-TAS method provides a new tool for studying PBM effects in other proteins and protein complexes that might respond to the CW wavelengths, such as Complex IV, in future studies.
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Láseres de Semiconductores , Espectrofotometría/métodos , Citocromos c/química , Cinética , Luz , Oxidación-ReducciónRESUMEN
Intense femtosecond pulse filamentation in open-air has been utilized for long distance optical communication and remote sensing, but it results in nonlinear-effect driven eye hazards which are not addressed by current eye safety standards. A systematic study of filamentation in atmospheric air was performed using a tunable 100 fs near-infrared laser (1100 nm-2400 nm). While undergoing filamentation, each source wavelength was spectrally broadened resulting in supercontinuum and third harmonic generation in the visible and near-IR spectrum. We record the spectra at the center and fringes of the supercontinuum as it is imaged onto a planar surface. In a full beam collection regime, we report the energy of the sub-1000 nm light generation for source wavelengths from 1100 nm to 1600 nm and compare the energy density to the maximum permissible exposure values under the ANSI Z136.1 laser safety standard.
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Polycrystalline zinc selenide (ZnSe) has been the subject of many nonlinear optics studies for wavelengths under 4.0 µm including sum/difference frequency generation, harmonic generation, and filamentation. In this report, the conversion efficiency of high harmonic generation (HHG) in ZnSe is quantified for mid-infrared wavelengths ranging from 2.7 µm to 8.0 µm. By increasing the fundamental wavelength, we demonstrate that HHG in thick ZnSe targets is limited by the band gap. The high conversion efficiency of mid-infrared to near-infrared light in ZnSe raises concerns of a nonlinear retinal hazard. We contrast the HHG behavior of ZnSe against the observed harmonic generation of calcium fluoride, BK7, and fused silica over the same wavelengths.
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Understanding the nonlinear properties of water is essential for laser surgery applications, as well as understanding supercontinuum generation in water. Unfortunately, the nonlinear properties of water for wavelengths longer than 1064 nm are poorly understood. We extend the application of the Z-scan technique in water to determine its nonlinear refractive index (n2) and nonlinear absorption (ß) for wavelengths in the 1150-1400 nm range, where linear absorption is also significant. We observe the wavelength-dependent variation of the nonlinear properties of water around the water absorption band.
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We perform a detailed theoretical and experimental investigation of supercontinuum generation in large-mode-area photonic crystal fibers pumped by a high-energy, high-repetition rate picosecond Nd:YVO4 laser, with the goal of using it as the Stokes beam in coherent anti-Stokes Raman scattering setup. We analyze the influence of fiber structure and length on the supercontinuum power, spectral shape, and group delay dispersion. We identify the experimental conditions for stable supercontinuum generation, with microjoule-level pulse energy and the spectrum extending beyond 1600 nm, which allows excitation of Raman frequencies up to 3000 cm-1 and beyond. We demonstrate reliable and efficient operation of a coherent anti-Stokes Raman spectroscopy and microscopy setup using this supercontinuum source.
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STUDY DESIGN: Descriptive. OBJECTIVES: To evaluate the physical demands of an international Rugby Union-level game using a global positioning system (GPS). BACKGROUND: Elite Rugby Union teams currently employ the latest technology to monitor and evaluate physical demands of training and games on their players. METHODS: GPS data from 2 players, a back and a forward, were collected during an international Rugby Union game. Locomotion speed, total body load, and body load sustained in tackles and scrums were analyzed. RESULTS: Players completed an average distance of 6715 m and spent the major portion of the game standing or walking, interspersed with medium- and high-intensity running activities. The back performed a higher number of high-intensity sprints and reached a greater maximal speed. Body load data revealed that high levels of gravitational force are sustained in tackling and scrum tasks. CONCLUSION: The current study provides a detailed GPS analysis of the physical demands of international Rugby Union players. These data, when combined with game video footage, may assist sports medicine professionals in understanding the demands of the game and mechanism of injury, as well as improving injury rehabilitation.