图像恢复问题中强度保持技术

提出了荧光造影图像序列的图像恢复方案。将新的强度约束合并到Miller正则化公式中,以获得保持高强度像素的能力。恢复的图像除了受光滑约束的影响之外,也会受到强度约束的影响。恢复过程中,采用从预滤波过程中获得的强度模板实现强度约束,该模板代表一幅由所期望强度值组成的图像。实验表明,该方案在高强度保持和图像恢复两方面都获得了较好的结果。     

Preparation and embedding–releasing behavior of polyelectrolyte microcapsule

以高压静电法制备的壳聚糖微球为模板, 与海藻酸钠层层自组装制备多层聚电解质膜, 再以乙酸溶出壳聚糖微球制备出微胶囊. 调节乙酸的浓度及溶出时间以控制核内壳聚糖的残留量, 从而诱导负电性荧光素钠的沉积. 微胶囊内部荧光素钠的浓度随着包埋时间的增加而呈线性地提高, 当荧光素钠溶液的浓度为2 mg/l时其最终包埋的浓度达到10.2 mg/l. 这种聚电解质微囊对于小分子负电荷荧光素钠的释放行为主要受其囊壁和内部模板结构的影响, 盐离子浓度对荧光素钠释放行为的影响较小.     

Mesoporous Aerogel Microparticles Injected into the Abdominal Cavity of Mice Accumulate in Parathymic Lymph Nodes

Mesoporous aerogel microparticles are promising drug delivery systems. However, their in vivo biodistribution pathways and health effects are unknown. Suspensions of fluorescein-labeled silica–gelatin hybrid aerogel microparticles were injected into the peritoneum (abdominal cavity) of healthy mice in concentrations of 52 and 104 mg kg⁻¹ in a 3-week-long acute toxicity experiment. No physiological dysfunctions were detected, and all mice were healthy. An autopsy revealed that the aerogel microparticles were not present at the site of injection in the abdominal cavity at the end of the experiment. The histological study of the liver, spleen, kidneys, thymus and lymphatic tissues showed no signs of toxicity. The localization of the aerogel microparticles in the organs was studied by fluorescence microscopy. Aerogel microparticles were not detected in any of the abdominal organs, but they were clearly visible in the cortical part of the parathymic lymph nodes, where they accumulated. The accumulation of aerogel microparticles in parathymic lymph nodes in combination with their absence in the reticuloendothelial system organs, such as the liver or spleen, suggests that the microparticles entered the lymphatic circulation. This biodistribution pathway could be exploited to design passive targeting drug delivery systems for flooding metastatic pathways of abdominal cancers that spread via the lymphatic circulation.     

Antiproliferative and Cytotoxic Activities of Fluorescein—A Diagnostic Angiography Dye

Fluorescein is a fluorescent dye used as a diagnostic tool in various fields of medicine. Although fluorescein itself possesses low toxicity, after photoactivation, it releases potentially toxic molecules, such as singlet oxygen (¹O₂) and, as we demonstrate in this work, also carbon monoxide (CO). As both of these molecules can affect physiological processes, the main aim of this study was to explore the potential biological impacts of fluorescein photochemistry. In our in vitro study in a human hepatoblastoma HepG2 cell line, we explored the possible effects on cell viability, cellular energy metabolism, and the cell cycle. We observed markedly lowered cell viability (≈30%, 75–2400 μM) upon irradiation of intracellular fluorescein and proved that this decrease in viability was dependent on the cellular oxygen concentration. We also detected a significantly decreased concentration of Krebs cycle metabolites (lactate and citrate < 30%; 2-hydroxyglutarate and 2-oxoglutarate < 10%) as well as cell cycle arrest (decrease in the G2 phase of 18%). These observations suggest that this photochemical reaction could have important biological consequences and may account for some adverse reactions observed in fluorescein-treated patients. Additionally, the biological activities of both ¹O₂ and CO might have considerable therapeutic potential, particularly in the treatment of cancer.     

Fluorescein Derivatives as Fluorescent Probes for pH Monitoring along Recent Biological Applications

Potential of hydrogen (pH) is one of the most relevant parameters characterizing aqueous solutions. In biology, pH is intrinsically linked to cellular life since all metabolic pathways are implicated into ionic flows. In that way, determination of local pH offers a unique and major opportunity to increase our understanding of biological systems. Whereas the most common technique to obtain these data in analytical chemistry is to directly measure potential between two electrodes, in biological systems, this information has to be recovered in-situ without any physical interaction. Based on their non-invasive optical properties, fluorescent pH-sensitive probe are pertinent tools to develop. One of the most notorious pH-sensitive probes is fluorescein. In addition to excellent photophysical properties, this fluorophore presents a pH-sensitivity around neutral and physiologic domains. This review intends to shed new light on the recent use of fluorescein as pH-sensitive probes for biological applications, including targeted probes for specific imaging, flexible monitoring of bacterial growth, and biomedical applications.     

Responsive Fluorophore Aggregation Provides Spectral Contrast for Fluorescence Lifetime Imaging

Fluorophores experience altered emission lifetimes when incorporated into and liberated from macromolecules or molecular aggregates; this trend suggests the potential for a fluorescent, responsive probe capable of undergoing self-assembly and aggregation and consequently altering the lifetime of its fluorescent moiety to provide contrast between the active and inactive probes. We developed a cyanobenzothioazole-fluorescein conjugate ( 1 ), and spectroscopically examined the lifetime changes caused by its reduction-induced aggregation in vitro. A decrease in lifetime was observed for compound 1 in a buffered system activated by the biological reducing agent glutathione, thus suggesting a possible approach for designing responsive self-aggregating lifetime imaging probes.     

Production of N-acetyl cis-4-hydroxy-L-proline by the yeast N-acetyltransferase Mpr1

We report that several tryptophan-rich peptides exhibit an affinity for a hydrophobic ionic liquid (IL) (1-ethyl-3-methylimidazolium bis-trifluoromethanesulfonyl imide), and that green fluorescent protein (GFP) fused to a peptides, "SSSWWSWWWW" (SW1) or "SWWWWSWWWW" (SW2), containing serine (S) and tryptophan (W) at the C terminus localized at the IL/water interface. While GFPs without W-rich peptide distributed only in water phase, SW1- and SW2-GFPs were accumulated at the interface. The localization of SW1-GFP showed biphasic behavior, and most distinctive localization was observed at 7.1 μM. The localization of SW2-GFP presumably occurred at largely lower concentration (≤0.5 μM) than that of SW1-GFP, which difference was due to the higher hydrophobicity of SW2 peptide.     

Development of Multicolor Flow Cytometry Calibration Standards: Assignment of Equivalent Reference Fluorophores (ERF) Unit

A procedure is described for assigning the number of equivalent reference fluorophores (ERF) values to microspheres labeled with a fluorophore designed to produce a fluorescence response in a given fluorescence channel of a multicolor flow cytometer. A fluorimeter was calibrated by a series of solutions of the reference fluorophores. The fluorimeter was used to obtain the microsphere fluorescence intensity, and a multicolor flow cytometer was used to obtain the microsphere concentration. The microsphere fluorescence intensity and the concentration were used to obtain the value of ERF for each model microsphere calibration standard. The procedure is described in detail only for microspheres with allophycocyanin (APC) immobilized on the surface. ERF values were also determined for microsphere calibrators for three other fluorescence channels: fluorescein isothiocyanate (FITC), phycoerythrin (PE), and Pacific Blue(PB). The four model microsphere calibrators provide a one point calibration for the four channels of a flow cytometer. By using software controls and changing the photomultiplier voltages, it is possible to obtain a multipoint calibration for each fluorescence channel using each microsphere calibrator.