Enhanced fluorescence from semiconductor quantum dot-labelled cells excited at 280 nm

Semiconductor quantum dots (QDs) have significant advantages over more traditional fluorophores used in fluorescence microscopy including reduced photobleaching, long-term photostability and high quantum yields, but due to limitations in light sources and optics, are often excited far from their optimum excitation wavelengths in the deep-UV. Here, we present a quantitative comparison of the excitation of semiconductor QDs at a wavelength of 280 nm, compared to the longer wavelength of 365 nm, within a cellular environment. We report increased fluorescence intensity and enhanced image quality when using 280 nm excitation compared to 365 nm excitation for cell imaging across multiple datasets, with a highest average fluorescence intensity increase of 3.59-fold. We also find no significant increase in photobleaching of QDs associated with 280 nm excitation.

Effectiveness of photon-initiated photoacoustic streaming in root canal models with different diameters or tapers

Background This study aimed to compare the use of photon-initiated photoacoustic streaming (PIPS) and conventional needle irrigation (CNI) in conjunction with different concentrations of sodium hypochlorite (NaOCl) to remove Enterococcus faecalis (E. faecalis) suspended bacteria and biofilms from root canal systems with different diameters or tapers. Methods Artificial root canal samples (n = 480) were randomly divided into three groups (n = 160/group). The canals were prepared to fit file sizes #10/.02, #25/.02, or #25/.06. The size #10/.02 group was incubated for seven days. The size #25/.02 or #25/.06 group was incubated for 2 days. A stable biological model of E. faecalis infection was established. The root canals were washed with distilled water or with 1%, 2%, or 5.25% NaOCl combined with CNI or PIPS. Bacterial suspensions and biofilms were assessed using an ATP assay kit and fluorescence microscopy. Image-Pro Plus was used to analyse the average fluorescence intensity to determine the most suitable root canal irrigation solution. Results In the CNI and PIPS groups, the ATP value of the 5.25% NaOCl subgroup was the lowest, followed by that of the 2% and 1% NaOCl subgroups. The ATP value of the distilled water subgroup was the highest (P < 0.05). When the root canal taper was 0.02, the ATP value of the #10/.02 + PIPS group was significantly lower than that of the #25/.02 + CNI group (P < 0.05). The average fluorescence intensity of the #10/.02 + PIPS group was lower than that of the #25/.02 + CNI group (P < 0.05). When the apical diameter was #25, the ATP value of the 0.02 taper in the PIPS group was lower than that of the 0.06 taper in the CNI group (P < 0.05), and the average fluorescence intensity of the 0.02 taper + PIPS group was lower than that of the 0.06 taper + CNI group (P < 0.05). PIPS combined with 2% and 5.25% NaOCl effectively improved the long-term antibacterial effect after irrigation and re-culture for 6 h. Conclusions Compared with CNI, PIPS has greater ability to remove bacteria in root canals with a small preparation diameter and a small taper. PIPS with 2% and 5.25% NaOCl exhibited superior antibacterial and bacteriostatic effects.

Analysis of Antiapoptosis Effect of Netrin-1 on Ischemic Stroke and Its Molecular Mechanism under Deleted in Colon Cancer/Extracellular Signal-Regulated Kinase Signaling Pathway

To analyze the regulatory effect of Netrin-1 in ischemic stroke and its influence on Deleted in Colon Cancer (DCC)/Extracellular Signal-regulated Kinase (ERK) signaling pathway, 20 male rats were selected to construct the rat model of middle cerebral artery occlusion (MCAO), 10 normal rats were selected as healthy controls (Normal Saline (NS)), and they were divided into the MCAO+Netrin-1 group, MCAO group, and NS group according to different treatment schemes. The positive expression of Netrin-1 was detected by immunostaining, magnetic resonance imaging (MRI) was adopted to detect the percentage of rat cerebral infarct volume in the cerebral hemispheres, and Modified Neurological Severity Score (mNSS) was adopted to evaluate postoperative neurological function in rats. Besides, a tunnel staining experiment was applied to detect the apoptosis rate of rat neurons, the sticker removal test was applied to evaluate the postoperative sensory function of rats, and fluorescence staining was adopted to detect the expression of DCC and ERK in rats. The results showed that the percentage of cerebral infarction volume in the cerebral hemispheres of the MCAO+Netrin-1 group was higher than that of the MCAO and NS groups ( P < 0.05 ); in the MCAO+Netrin-1 group, the MCAO mNSS scoring and the time spent in the sticker removal test were lower than the MCAO group ( P < 0.05 ); the apoptosis rate of rats in the MCAO+Netrin-1 group was lower than that in the MCAO group ( P < 0.05 ); the average fluorescence intensity of DCC and p-ERK in the MCAO+Netrin-1 group was higher than that in the MCAO group ( P < 0.05 ); the average fluorescence intensity of p-ERK in the MCAO+Netrin-1 group was higher than that in the MCAO group ( P < 0.05 ). In short, Netrin-1 can effectively reduce the brain tissue damage in rats with ischemic stroke, improve the nerve function and sensory function of rats, and inhibit neuronal cell apoptosis. Netrin-1 can promote DCC expression and ERK phosphorylation, and the EPK signaling pathway may be involved in the antiapoptotic effect of Netrin-1.

Visualisation of Cornstarches Granule with Different Amylose/Amylopectin Contents: A Combined CLSM and SEM Approach

The morphologies and microstructures of cornstarches with different amylose/amylopectin ratios (waxy: 0/100; maize: 23/77; Gelose 50: 50/50; Gelose 80: 80/20) were studied by a combined SEM and confocal laser scanning microscope (CLSM). CLSM allowed the visualization of cross-sections of starch granules without the need for sectioning techniques that lead to destruction of the microstructure of sample. The influence of the amylose/amylopectin ratio on the internal structures and morphologies was able to be revealed by this choice of techniques. It was found that the granules of the high-amylopectin starches were more regular in shape than those of the high-amylose starches, but the surfaces of the high-amylose starches were smoother than those of the high-amylopectin starches. The high-amylose starches (G50 and G80) were brighter than that of low-amylopectin starches (waxy and maize) under confocal laser light, and the average fluorescence intensity sequence of the granules was G80>G50>maize>waxy. Waxy and maize starches show clearly cavities and channels while G50 and G80 present bright core.

The effect of 2,5-hexanedione on permeability of blood-nerve barrier in rats

To explore the effect of 2,5-hexanedione on permeability of blood-nerve barrier, adult Wistar rats were administered with 400 mg.kg—1.d— 1 2,5-hexanedione to establish animal model of 2,5-hexnedione neuropathy. Evans blue was injected through left femoral vein of the rats after the model had been established. The distribution of fluorescence in sciatic-tibial nerve was observed and assessed. For the transverse sections of sciatic-tibial nerves, the average fluorescence intensity of proximal section was stronger (p < .01) than those of intermediate and distal sections and the average fluorescence intensity of intermediate section was stronger (p < .01) than that of distal section in the intoxicated group. In the control, the weak fluorescence was shown, and average fluorescence intensity of distal section was stronger (p < .05) than that of proximal section. The average fluorescence intensity of proximal, intermediate and distal sections in the intoxicated group was stronger (p < .01) than those of the corresponding sections in the control. For the longitudinal sections of sciatic-tibial nerves, fluorescence was observed in both proximal and distal sections in the intoxicated group. The fluorescence intensity of distal section in the control was weak and almost no fluorescence was shown in the proximal section. The permeability of blood-nerve barrier could be increased by 2,5-hexanedione.

Optimization of the Self-Quenching Response of Nitrobenzoxadiazole Dipalmitoylphosphatidylethanolamine in Phospholipid Membranes for Biosensor Development

Incorporation of the lipid-conjugated fluorescent probe nitrobenzoxadiazole dipalmitoylphosphatidylethanolamine (NBD-PE) into bilayer lipid membranes (BLMs) provides a matrix wherein changes in the structure of the membrane can be transduced into changes in fluorescence intensity or lifetime. In the work reported here, a comparison was made between an empirical model recently developed by our group to account for alterations in the fluorescence lifetime and average fluorescence intensity of NBD-PE as a result of self-quenching and an earlier alternative model which describes self-quenching of membrane-bound chlorophyll a. Our model showed the more satisfactory correlation with self-quenching data obtained from lipid membranes containing 1 to 50 mol % of NBD-PE. This model was used to determine the optimum initial surface concentration of NBD-PE to be incorporated into phospholipid membranes for biosensor development. Optimization was based on the magnitude of the change in fluorescence intensity as a function of changes in the local concentration of the probe. The presence of acidic headgroups in the membrane results in negligible improvement in sensitivity, while a heterogeneous membrane structure greatly enhances the signal magnitude. Experimental results did not provide accurate optimum concentrations, although two NBD-PE surface concentrations were found to yield close agreement with theoretically predicted optimum surface concentrations of 0.027 ± 0.001 and 0.073 ± 0.001 molecules NBD-PE nm−2.