Enhanced lateral resolution in continuous wave stimulated emission depletion microscopy using tightly focused annular radially polarized excitation beam.

The lateral resolution of continuous wave (CW) stimulated emission depletion (STED) microscopy is enhanced about 12% by applying annular-shaped amplitude modulation to the radially polarized excitation beam. A focused annularly filtered radially polarized excitation beam provides a more condensed point spread function (PSF), which contributes to enhance effective STED resolution of CW STED microscopy. Theoretical analysis shows that the FWHM of the effective PSF on the detection plane is smaller than for conventional CW STED. Simulation shows the donut-shaped PSF of the depletion beam and confocal optics suppress undesired PSF sidelobes. Imaging experiments agree with the simulated resolution improvement.

Dual-wavelength Fourier ptychography using a single LED.

We propose dual-wavelength Fourier ptychography for topographic measurement. To extend the axial measurement range, a single light-emitting diode (LED) and two appropriate bandpass filters are employed. This provides a speckle-free phase image, and reduces the possibility of a systematic error, which yields a high-quality topographic image. The proposed system can measure the surface topography in the range of nano- to micro-structures. The performance of the system is experimentally verified.

Investigation on improvement of lateral resolution of continuous wave STED microscopy by standing wave illumination.

In this paper, we report the enhancement of resolution of continuous wave (CW) stimulated emission depletion (STED) microscopy by a novel method of structured illumination of an excitation beam. Illumination by multiple excitation beams through the specific pupil apertures with high in-plane wave vectors leads to interference of diffracted light flux near the focal plane, resulting in the contraction of the point spread function (PSF) of the excitation. Light spot reduction by the suggested standing wave (SW) illumination method contributes to make up much lower depletion efficiency of the CW STED microscopy than that of the pulsed STED method. First, theoretical analysis showed that the full width at half maximum (FWHM) of the effective PSF on the detection plane is expected to be smaller than 25% of that of conventional CW STED. Second, through the simulation, it was elucidated that both the donut-shaped PSF of the depletion beam and the confocal optics suppress undesired contribution of sidelobes of the PSF by the SW illumination to the effective PSF of the STED system. Finally, through the imaging experiment on 40-nm fluorescent beads with the developed SW-CW STED microscopy system, we obtained the result which follows the overall tendency from the simulation in the aspects of resolution improvement and reduction of sidelobes. Based on the obtained result, we expect that the proposed method can become one of the strategies to enhance the resolution of the CW STED microscopy.

Retrograde balloon dilation as a therapeutic option for post-gynecologic surgery ureteral stricture followed by ureteroureterostomy: a comparative study regarding stricture length.

Background: To evaluate the success rate of balloon dilation and the factors possibly influencing the outcomes of balloon dilation for the ureteric strictured portion of ureteroureterostomy (UUS) site in patients with post-gynecologic surgeries. Methods: A single institution data base was screened for the patients who received balloon dilation for a treatment of ureteral stricture diagnosed after gynecologic surgery. Overall 114 patients underwent primary intra-operative UUS due to ureteral injury during gynecologic surgery. Among them, 102 patients received balloon dilation, and their medical records were retrospectively reviewed. Success of balloon dilation was defined as the condition that requires no further clinical interventions after 6 months from balloon dilation. Results: The ureter injury rate of women treated with open radical abdominal hysterectomy was highest (32 cases, 31.4%). 60 patients (60.8%) showed successful outcomes regarding dilation. All patients underwent technically successful dilation with a full expansion of balloon during the procedure, but 40 patients (39.2%) were clinically unsuccessful as they showed a recurrence of ureteral stricture on the previous balloon dilation site after the first dilation procedure. Univariate logistic regression analyses showed that stricture length >2 cm was a significant predictor of successful dilation (odds ratio, 0.751; 95% confidence interval, 0.634-0.901; p-value, 0.030), but it failed to achieve independent predictor status in multivariate analysis. Conclusion: Balloon dilation can an effective alternative treatment option for strictured portion of the primary UUS in post-gynecologic surgery patients when its length is <2 cm.

Investigation on achieving super-resolution by solid immersion lens based STED microscopy.

The feasibility of stimulated emission depletion (STED) microscopy using a solid immersion lens was investigated. First, the theoretical feasibility of the considered system is discussed based on a vectorial field algorithm that uses a stratified medium composed of a SIL air-gap and test sample. Using the simulation, we verified that evanescent waves with much higher spatial frequencies corresponding to the high numerical aperture in the air-gap can be utilized to achieve a higher resolution than a confocal fluorescent image without a depletion beam. The simulated expectation was supported by actual imaging on two types of samples: fluorescent beads with a 20 nm diameter and an actin sample with a filamentous structure. The lateral resolution of the system was determined to be 34 nm via the imaging results on the nano-beads. The system was quite promising for achieving nano-scale surface imaging of biological samples; an even higher resolution was achieved by adjusting the wavelength and the intensity of the depletion beam.

Ethylene vinyl acetate copolymer/multi-walled carbon nanotube nanocomposites.

In this study relatively large amount of ethylene-vinyl acetate copolymer (EVA) (130 g) was melt-mixed with multi walled carbon nanotube (MWCNT) and the effect of mixing time and rotor speed on electrical properties were investigated. Also, to investigate the relationship between the degree of dispersion of MWCNT and conductivity, the dispersion of MWCNT was examined using SEM and TEM. At shorter mixing time (15 min), increased rotor speed produces improved conductivity, whereas at longer mixing time (25 min and 35 min) increased rotor speed leads to enhanced breakage of the MWCNT and decrease of conductivity even though the dispersion of MWCNT is improved. Therefore to obtain the best conductivity, optimal shear rate and mixing time should be chosen.

Imipramine inhibits A-type delayed rectifier and ATP-sensitive K+ currents independent of G-protein and protein kinase C in murine proximal colonic myocytes.

The effects of imipramine on A-type delayed rectifier K+ currents and ATP-sensitive K+ (KATP) currents were studied in isolated murine proximal colonic myocytes using the whole-cell patch-clamp technique. Depolarizing test pulses between -80 mV and +30 mV with 10 mV increments from the holding potential of -80 mV activated voltage-dependent outward K+ currents that peaked within 50 ms followed by slow decreasing sustained currents. Early peak currents were inhibited by the application of 4-aminopyridine, whereas sustained currents were inhibited by the application of TEA. The peak amplitude of A-type delayed rectifier K+ currents was reduced by external application of imipramine. The half-inactivation potential and the half-recovery time of A-type delayed rectifier K+ currents were not changed by imipramine. With 0.1 mM ATP and 140 mM K+ in the pipette and 90 mM K+ in the bath solution and a holding potential of -80 mV, pinacidil activated inward currents; this effect was blocked by glibenclamide. Imipramine also inhibited KATP currents. The inhibitory effects of imipramine in A-type delayed rectifier K+ currents and KATP currents were not changed by guanosine 5-O-(2-thiodiphosphate) (GDPbetaS) and chelerythrine, a protein kinase C inhibitor. These results suggest that imipramine inhibits A-type delayed rectifier K+ currents and KATP currents in a manner independent of G-protein and protein kinase C.

Action of imipramine on activated ATP-sensitive K(+) channels in interstitial cells of Cajal from murine small intestine.

Tricyclic antidepressants have been widely used for the treatment of depression and as a therapeutic agent for the altered gastrointestinal (GI) motility of irritable bowel syndrome (IBS). The aim of this study was to clarify whether antidepressants directly modulate pacemaker currents in cultured interstitial cells of Cajal (ICC). We used the whole-cell patch-clamp techniques at 30 degrees C in cultured ICC from the mouse small intestine. Treatment of pinacidil, an ATP-sensitive K(+) channel opener, in the ICC using the current clamping mode, produced hyperpolarization of the membrane potential and decreased the amplitude of the pacemaker potentials. With the voltage clamp mode, we observed a decrease in the frequency and amplitude of pacemaker currents and increases in the resting outward currents. These effects of pinacidil on pacemaker potentials and currents were completely suppressed by glibenclamide, an ATP-sensitive K(+) channel blocker. Also, with the current clamp mode, imipramine blocked the affect of pinacidil on the pacemaker potentials. Observations of the voltage clamp mode with imipramine, desipramine and amitryptyline suppressed the action of pinacidil in the ICC. Next, we examined whether protein kinase C (PKC) and the G protein are involved in the action of imipramine on pinacidil induced pacemaker current inhibition. We used chelerythrine, a potent PKC inhibitor and GDPbetaS, a nonhydrolyzable guanosine 5-diphosphate (GDP) analogue that permanently inactivates GTP-binding proteins. We found that pretreatment with chelerythrine and intracellular application of GDPbetaS had no influence on the blocking action of imipramine on inhibited pacemaker currents by pinacidil. We conclude that imipramine inhibited the activated ATP-sensitive K(+) channels in ICC. This action does not appear to be mediated through the G protein and protein kinase C. Furthermore, this study may suggest another possible mechanism for tricyclic antidepressants related modulation of GI motility.

Influence of naloxone on catecholamine release evoked by nicotinic receptor stimulation in the isolated rat adrenal gland.

The present study was designed to investigate the effect of naloxone, a well known opioid antagonist, on the secretion of catecholamines (CA) evoked by cholinergic stimulation and membrane-depolarization in the isolated perfused rat adrenal glands, and to establish its mechanism of action. Naloxone (10(-6) approximately 10(-5) M), perfused into an adrenal vein for 60 min, produced dose- and time-dependent inhibition of CA secretory responses evoked by ACh (5.32 x 10(-3) M), high K+ (5.6 x 10(-2) M), DMPP (10(-4) M) and McN-A-343 (10(-4) M). Naloxone itself also failed to affect the basal CA output. In adrenal glands loaded with naloxone (3 x 10(-6) M), the CA secretory responses evoked by Bay-K-8644, an activator of L-type Ca2+ channels, and cyclopiazonic acid, an inhibitor of cytoplasmic Ca(2+)-ATPase, were also inhibited. In the presence of met-enkephalin (5 x 10(-6) M), a well known opioid agonist, the CA secretory responses evoked by ACh, high K+, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were also significantly inhibited. Taken together, these results suggest that naloxone greatly inhibits the CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as that by membrane depolarization. It seems that these inhibitory effects of naloxone does not involve opioid receptors, but might be mediated by blocking both the calcium influx into the rat adrenal medullary chromaffin cells and the uptake of Ca2+ into the cytoplasmic calcium store, which are at least partly relevant to the direct interaction with the nicotinic receptor itself.