Single-Molecule-Sensitive FRET in Freely-Diffusing Attoliter Droplets

Fluorescence resonance energy transfer (FRET) from individual, dye-labeled RNA molecules confined in freely-diffusing attoliter-volume aqueous droplets is carefully compared to FRET from unconfined RNA in solution. The use of freely-diffusing droplets is a remarkably simple and high-throughput technique that facilitates a substantial increase in signal-to-noise for single-molecular-pair FRET measurements. We show that there can be dramatic differences between FRET in solution and in droplets, which we attribute primarily to an altered pH in the confining environment. We also demonstrate that a sufficient concentration of a non-ionic surfactant mitigates this effect and restores FRET to its neutral-pH solution value. At low surfactant levels, even accounting for pH, we observe differences between the distribution of FRET values in solution and in droplets which remain unexplained. Our results will facilitate the use of nanoemulsion droplets as attoliter volume reactors for use in biophysical and biochemical assays, and also in applications such as protein crystallization or nanoparticle synthesis, where careful attention to the pH of the confined phase is required.Comment: Twenty-three pages, 3 figures, one table, plus supporting information with additional figures and table

Hydrosomes: femtoliter containers for fluorescence spectroscopy studies

We report on improvements and innovations in the use of hydrosomes to encapsulate and study single molecules. Hydrosomes are optically-trappable aqueous nanodroplets. The droplets are suspended in a fluorocarbon medium that is immiscible with water and has an index of refraction lower than water, so hydrosomes are stable and optically trapped by a focused laser beam (optical tweezers). Using optical tweezers, we hold the hydrosomes within a confocal observation volume and interrogate the encapsulated molecule by fluorescence excitation. This method allows for long observation times of a molecule without the need for surface immobilization or liposome encapsulation. We have developed a new way for creating hydrosomes on demand by inertially launching them into the fluorocarbon matrix using a piezo-activated micropipette. Time-resolved fluorescence anisotropy studies are carried out to characterize the effects of the hydrosome interface boundary on biological molecules and to determine whether molecules encapsulated within hydrosomes diffuse freely throughout the available volume. We measured the fluorescence anisotropy decay of 20mer DNA duplexes, and enhanced green fluorescent protein (GFP). We conclude that the molecules rotate freely inside the nanodroplets and do not stick or aggregate at the boundary

Hydrosomes: femtoliter containers for fluorescence spectroscopy studies

We report on improvements and innovations in the use of hydrosomes to encapsulate and study single molecules. Hydrosomes are optically-trappable aqueous nanodroplets. The droplets are suspended in a fluorocarbon medium that is immiscible with water and has an index of refraction lower than water, so hydrosomes are stable and optically trapped by a focused laser beam (optical tweezers). Using optical tweezers, we hold the hydrosomes within a confocal observation volume and interrogate the encapsulated molecule by fluorescence excitation. This method allows for long observation times of a molecule without the need for surface immobilization or liposome encapsulation. We have developed a new way for creating hydrosomes on demand by inertially launching them into the fluorocarbon matrix using a piezo-activated micropipette. Time-resolved fluorescence anisotropy studies are carried out to characterize the effects of the hydrosome interface boundary on biological molecules and to determine whether molecules encapsulated within hydrosomes diffuse freely throughout the available volume. We measured the fluorescence anisotropy decay of 20mer DNA duplexes, and enhanced green fluorescent protein (GFP). We conclude that the molecules rotate freely inside the nanodroplets and do not stick or aggregate at the boundary

Perbedaan Asupan Lemak, Lingkar Pinggang dan Persentase Lemak Tubuh pada Wanita Dislipidemia dan Non Dislipidemia

Differences of fat intake, waist circumference and percentage of bodt fat in dyslipidemia and non dyslipidemia adult women: Heart disease is the leading cause of death in several countries in the world . One of the major risk factors for heart disease is dyslipidemia . Dyslipidemia is a disorder of lipid metabolism characterized by an increase or decrease in plasma lipid fractions . Dyslipidemia has a strong relationship with the occurrence of central obesity . The purpose of this study was to analyze differences in the intake of fat , waist circumference and body fat percentage in dyslipidemia and non dyslipidemia adult women. This research is analytic study with cross sectional approach . The population in this study were adult women who examined their lipid profile in December 2013 in the Clinical Laboratory Cito Indraprasta Semarang . The total sample was 32 people . Independent test analysis of the differences using t-test for variables waist circumference and Mann Whitney test for variable fat intake and body fat percentage to 95 % and a significance level of 5% error. The results showed 17 adult women ( 53.1 % ) and 15 female adult dyslipidemia ( 46.9 % ) non- dyslipidemia. Average intake of fat, waist circumference and percentage body fat in adult women dyslipidemia higher than non dyslipidemia in adult women. Analysis of statistical tests showed difference in fat intake , waist circumference and body fat percentage in women adult dyslipidemia and non dyslipidemia (p value, respectively p = 0.002, p = 0.0001 and p = 0.0001

Theory of imaging a photonic crystal with transmission near-field optical microscopy

While near-field scanning optical microscopy (NSOM) can provide optical images with resolution much better than the diffraction limit, analysis and interpretation of these images is often difficult. We present a theory of imaging with transmission NSOM that includes the effects of tip field, tip/sample coupling, light propagation through the sample and light collection. We apply this theory to analyze experimental NSOM images of a nanochannel glass (NCG) array obtained in transmission mode. The NCG is a triangular array of dielectric rods in a dielectric glass matrix with a two-dimensional photonic band structure. We determine the modes for the NCG photonic crystal and simulate the observed data. The calculations show large contrast at low numerical aperture (NA) of the collection optics and detailed structure at high NA consistent with the observed images. We present calculations as a function of NA to identify how the NCG photonic modes contribute to and determine the spatial structure in these images. Calculations are presented as a function of tip/sample position, sample index contrast and geometry, and aperture size to identify the factors that determine image formation with transmission NSOM in this experiment.Comment: 28 pages of ReVTex, 14 ps figures, submitted to Phys. Rev.

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection