Note: Time-gated 3D single quantum dot tracking with simultaneous spinning disk imaging.

We describe recent upgrades to a 3D tracking microscope to include simultaneous Nipkow spinning disk imaging and time-gated single-particle tracking (SPT). Simultaneous 3D molecular tracking and spinning disk imaging enable the visualization of cellular structures and proteins around a given fluorescently labeled target molecule. The addition of photon time-gating to the SPT hardware improves signal to noise by discriminating against Raman scattering and short-lived fluorescence. In contrast to camera-based SPT, single-photon arrival times are recorded, enabling time-resolved spectroscopy (e.g., measurement of fluorescence lifetimes and photon correlations) to be performed during single molecule/particle tracking experiments.

Effects of nifedipine on esophageal tone and perception of esophageal distension.

The effects of nifedipine (20 mg orally) on esophageal body resting tone and perception of esophageal distension were compared with those of placebo in 10 healthy volunteers, using a double-blind crossover design. A 3-cm silicon balloon positioned 10 cm above the lower esophageal sphincter was inflated with 2-20 ml of air, in 2-ml increments. The subjects scored their chest sensations, while pressure in the balloon was continuously recorded. In each experiment three series of measurements were made, at baseline and 15 and 30 min after administration of nifedipine or placebo. Bench tests were run to determine pressure-to-volume relationships for each of the balloons used in the study. Thirty minutes after nifedipine ingestion, a significant decrease in systolic blood pressure (from 112.6 +/- 2.3 to 99.0 +/- 2.0 mm Hg) was observed. Amplitude and duration of esophageal peristaltic contractions were significantly decreased by nifedipine (from 128.1 +/- 16.7 to 98.7 +/- 10.6 mm Hg and from 3.9 +/- 0.3 to 3.3 +/- 0.2 sec, respectively). With balloon volumes of 8 ml and higher, balloon pressures were higher with the balloon in the esophagus than on the bench, the difference being determined by the compliance of the esophagus. This difference rose from 18.5 +/- 6.8 to 40.2 +/- 7.2 mm Hg (8-20 ml) with placebo and from 23.1 +/- 3.7 to 35.9 +/- 5.8 mm Hg with nifedipine. No significant differences between nifedipine and placebo were found. The esophageal sensation scores increased linearly with increasing balloon volume from 6 ml onwards. Nifedipine had no significant effect on the perception of esophageal balloon distension.

Cerebral potentials evoked by oesophageal distension in patients with non-cardiac chest pain.

Cerebral evoked potential recording was used to study the oesophagus-brain axis in 10 controls and 10 patients with non-cardiac chest pain with a manometric diagnosis of either nutcracker oesophagus or diffuse spasm and a positive edrophonium test. A series of 50 inflations (10/minute; inflation rate of 170 ml/second) of an intraoesophageal balloon (5 cm proximal to the lower oesophageal sphincter) was performed in each subject. Three different inflation volumes were used and were individually determined to cause no sensation, slight sensation, and definite sensation, respectively (volume ranges: 2-8 ml, 5-18 ml, and 8-22 ml). All signals were coded and their quality was scored on a scale from 0 (no recognisable pattern) to 5 (well defined potential of good quality) by four 'blinded' observers. The evoked potential quality scores and amplitude of the major peaks increased significantly (p less than 0.01) with increasing sensation, both in patients and in controls. In the patients, quality score and amplitude of all four peaks of the evoked potentials were lower (p less than 0.05) and latencies of two of the four peaks were longer (p less than 0.02) than in the controls. The volumes of air required to produce the various sensations were lower in the patients (p less than 0.01). When divided by the balloon volume, amplitude and quality of the evoked potential were no longer significantly different between the groups. These results suggest that the increased perception of oesophageal distension in patients with non-cardiac chest pain is caused by altered central processing rather than (functionally) abnormal receptors in the oesophageal wall.

Cerebral potentials evoked by esophageal distension in human.

Recent studies have shown that cerebral evoked potentials (EPs) can be recorded after balloon distension of the human esophagus. The aim of this study was to evaluate the characteristics of these viscerosensory EPs and to investigate the relations between these EPs and perception of esophageal distension. Nineteen healthy volunteers (22-60 yr old) were studied. A balloon positioned 5 cm above the lower esophageal sphincter was inflated 10 times each minute. EPs recorded from four midline scalp electrodes were averaged for 50 and 100 inflation cycles. A clearly defined triphasic (negative-positive-negative) EP was recorded in all subjects and from all four recording sites when a volume leading to definite sensation and rapid balloon inflation (170 ml/s) was used. The latencies of the peaks were 231 +/- 7 (N1), 303 +/- 7 (P1), and 379 +/- 8 ms (N2). No significant correlation between stimulus perception and latency was found. The amplitude and quality of the EPs (scored by 3 blinded observers) increased significantly (P less than 0.01, ANOVA) with increasing sensation. Slow balloon inflation (30 ml/s) was significantly less effective in evoking EPs than rapid inflation (P less than 0.01). The EPs evoked by 100 inflations were not significantly clearer than those evoked by 50 inflations. It is concluded that the ability to record cerebral potentials evoked by esophageal balloon distension is related to the rate of balloon inflation and to the level of awareness of the stimulus.