Gold nanoparticles are efficiently labelled with a luminescent ruthenium complex, producing 13 and 100 nm diameter, monodisperse red-emissive imaging probes with luminescence lifetimes prolonged over the molecular unit. Single, 100 nm particles are observed in whole cell luminescence imaging which reveals their biomolecular association with chromatin in the nucleus of cancer cells.
The development of long-lived luminescent nanoparticles for lifetime imaging is of wide interest as luminescence lifetime is environmentally sensitive detection independent of probe concentration. We report novel iridium-coated gold nanoparticles as probes for multiphoton lifetime imaging with characteristic long luminescent lifetimes based on iridium luminescence in the range of hundreds of nanoseconds and a short signal on the scale of picoseconds based on gold allowing multichannel detection. The tailor-made IrC complex forms stable, water-soluble gold nanoparticles (AuNPs) of 13, 25, and 100 nm, bearing 1400, 3200, and 22 000 IrC complexes per AuNP, respectively. The sensitivity of the iridium signal on the environment of the cell is evidenced with an observed variation of lifetimes. Clusters of iridium nanoparticles show lifetimes from 450 to 590 ns while lifetimes of 660 and 740 ns are an average of different points in the cytoplasm and nucleus. Independent luminescence lifetime studies of the nanoparticles in different media and under aggregation conditions postulate that the unusual long lifetimes observed can be attributed to interaction with proteins rather than nanoparticle aggregation. Total internal reflection fluorescence microscopy (TIRF), confocal microscopy studies and 3D luminescence lifetime stacks confirm the presence of bright, nonaggregated nanoparticles inside the cell. Inductively coupled plasma mass spectrometry (ICPMS) analysis further supports the presence of the nanoparticles in cells. The iridium-coated nanoparticles provide new nanoprobes for lifetime detection with dual channel monitoring. The combination of the sensitivity of the iridium signal to the cell environment together with the nanoscaffold to guide delivery offer opportunities for iridium nanoparticles for targeting and tracking in in vivo models.
Cutting of bone with ultrasonics is influenced by the load applied and the setting used. Care must be used to prevent the tip from sliding over the bone at low loadings.
Aim:Imaging of blood flow in narrow channels and close to vessel walls is important in cardiovascular research for understanding pathogenesis. Our aim was to provide novel nanoprobes with visible emission and long lifetimes as trackers of flow. Materials & methods: Gold nanoparticles coated with an iridium complex were prepared. Luminescence imaging was used to monitor their flows in different hematocrit blood and in murine tissues. Results: The velocities are independent of hematocrit level and the nanoparticles entering blood circulation can be clearly detected in vessels in lungs, mesentery and the skeletal muscle. Conclusion: The work introduces for the first time iridium-based yellow-green luminescence with nanoparticle size of 100 nm for visualizing and monitoring flows with much higher resolution than conventional alternatives.
Optical microscopy
techniques are ideal for live cell imaging for
real-time nanoparticle tracking of nanoparticle localization. However,
the quantification of nanoparticle uptake is usually evaluated by
analytical methods that require cell isolation. Luminescent labeling
of gold nanoparticles with transition metal probes yields particles
with attractive photophysical properties, enabling cellular tracking
using confocal and time-resolved microscopies. In the current study,
gold nanoparticles coated with a red-luminescent ruthenium transition
metal complex are used to quantify and track particle uptake and localization.
Analysis of the red-luminescence signal from particles is used as
a metric of cellular uptake, which correlates to total cellular gold
and ruthenium content, independently measured and correlated by inductively
coupled plasma mass spectrometry. Tracking of the luminescence signal
provides evidence of direct diffusion of the nanoparticles across
the cytoplasmic membrane with particles observed in the cytoplasm
and mitochondria as nonclustered “free” nanoparticles.
Electron microscopy and inhibition studies identified macropinocytosis
of clusters of particles into endosomes as the major mechanism of
uptake. Nanoparticles were tracked inside GFP-tagged cells by following
the red-luminescence signal of the ruthenium complex. Tracking of
the particles demonstrates their initial location in early endosomes
and, later, in lysosomes and autophagosomes. Colocalization was quantified
by calculating the Pearson’s correlation coefficient between
red and green luminescence signals and confirmed by electron microscopy.
Accumulation of particles in autophagosomes correlated with biochemical
evidence of active autophagy, but there was no evidence of detachment
of the luminescent label or breakup of the gold core. Instead, accumulation
of particles in autophagosomes caused organelle swelling, breakdown
of the surrounding membranes, and endosomal release of the nanoparticles
into the cytoplasm. The phenomenon of endosomal release has important
consequences for the toxicity, cellular targeting, and therapeutic
future applications of gold nanoparticles.
The use of silica sub-micron particles on hard dentine tissues is dependent on the modification of the surface coating of the particles. This may influence how particles are incorporated in potential delivery vehicles applied to the dentine surface with the employment of a fluorosurfactant showing promise.
Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) in poultry is most often transmitted by the fecal− oral route, which can be attributed to high population density. Upon encountering the innate immune response in a host, the pathogen triggers a stress response and virulence factors to help it survive in the host. The aim of this study was to evaluate the effect of hypromellose acetate/succinate (HPMCAS)-coated alginate microparticles containing the Ctx(Ile 21 )-Ha antimicrobial peptide (AMP) on both intestinal colonization and systemic infection of laying hens challenged with S. Enteritidis. The applied AMP microsystem reduced the bacterial load of S. Enteritidis in the liver, with a statistical significance between groups A (control, no Ctx(Ile 21 )-Ha peptide) and B (2.5 mg of Ctx(Ile 21 )-Ha/kg) at 2 days postinfection (dpi), potentially indicating the effectiveness of Ctx(Ile 21 )-Ha in the first stage of infection by S. Enteritidis. In addition, the results showed a significant decrease in the S. Enteritidis counts in the spleen and cecal content at 5 dpi; remarkably, no S. Enteritidis counts were observed in livers at 5, 7, and 14 dpi, regardless of the Ctx(Ile 21 )-Ha dosage (p-value <0.0001). Using the Chi-square test, the effect of AMP microparticles on S. Enteritidis fecal excretion was also evaluated, and a significantly lower bacterial excretion was observed over 21 days in groups B and C, in comparison with the untreated control (p-value <0.05). In summary, the use of HPMCAS-Ctx(Ile 21 )-Ha peptide microcapsules in laying hens drastically reduced the systemic infection of S. Enteritidis, mainly in the liver, indicating a potential for application as a feed additive against this pathogen.
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