Fluorescent semiconductor nanocrystals (q-dots) benefit from practical features such as high fluorescence intensity, broad
excitation band and emission diameter dependency. These unique spectroscopic characterizations make q-dots excellent candidates
for new fluorescent labels in multi-chromatic analysis, such as Flow-Cytometry (FCM). In this work we shall present new possibilities
of multi-labeling and multiplex analysis of pathogenic bacteria, by Flow-Cytometry (FCM) analysis and new specific IgG—q-dots
conjugates. We have prepared specific conjugates against B. anthracis spores (q-dots585-IgGαB. anthracis and q-dots655-IgGαB.anthracis). These conjugates enabled us to achieve double staining of B. anthracis spores which improve the FCM analysis specificity versus control Bacillus spores. Moreover, multiplexed analysis of B. anthracis spores and Y. pestis bacteria was achieved by using specific antibodies labeled with different q-dots to obtain: q-dots585-IgGαB. anthracis and q-dots655-IgGαY.pestis, each characterized by its own emission peak as a marker. Specific and sensitive multiplex analysis for both pathogens has
been achieved, down to 103 bacteria per ml in the sample. 相似文献
We report the first simultaneous measurement of surface-confined and solution fluorescence correlation spectroscopy (FCS). We use an optical configuration for tightly focused excitation and separate detection of light emitted below (undercritical angle fluorescence, UAF) and above (supercritical angle fluorescence, SAF) the critical angle of total internal reflection of the coverslip/sample interface. This creates two laterally coincident detection volumes which differ in their axial extent. While detection of far-field UAF emission producesa standard confocal volume, near-field-mediated SAF produces a highly surface-confined detection volume at the coverslip/sample interface which extends only ~200 nm into the sample. A characterization of the two detection volumes by FCS of free diffusion is presented and compared with analytical models and simulations. The presented FCS technique allows to determine bulk solution concentrations and surface-near concentrations at the same time. 相似文献
The present study evaluates the photodynamic damage with 5-aminolevulinic acid (5-ALA) using HeLa as experimental model. HeLa
cell line was irradiated with red light (He-Ne laser, λ = 632.8 CW nm). The influence of different incubation times and concentrations
of 5-ALA, different irradiation doses and various combinations of photosensitizer and light doses on the cellular viability
of HeLa cells were studied. The optimal uptake of photosensitizer ALA in HeLa cells was investigated by means of PpIX fluorescence
intensity by exciting the HeLa cell suspension at 450 nm and a detection wavelength set at 690 nm. Cells viability was determined
by means of trypan blue solution. The spectrometric measurements showed that the maximal cellular uptake of 5-ALA occurred
after 4 h in vitro incubation. We found that the combination with 5-ALA and laser irradiation leads to time/concentration-dependent
increase of cells death and also energy doses-dependent enlarge the cells death. The fluorescence intensity after PDD of carcinoma
cells reduce when compared with the control group. The fluorescence emission spectral profiles after PDD of carcinoma cells
showed a dip around 425–525 nm when compared with the control group. This may be due to the damage of mitochondria component
of cells. The percentage of HeLa cells after PDD shows that the percentage of cells survival rate as function of laser dose
(power). Hence it is clear that at 200 μg/ml ALA and 20 mW laser irradiation, more than 70% of HeLa cells were dead after
15 min. 相似文献
Strong surface (metal) enhanced fluorescence (SEF or MEF) is observed from clusters and single E coli bacteria cells labeled with Carbon nanodots (CDs), which were synthesized from date pits. The enhancement factor (EF) for SEF of the cell clusters were close to 50 for both 533 and 633 nm laser excitation wavelength. Those EFs are ratios of emission peak areas from CD labeled cell clusters on gold film to the peak areas of the same batch cell clusters on glass substrate. SEF with 633 nm excitation performed better than SEF with 532 nm excitation, achieving higher fluorescence intensity and much higher contrast. The contrast as high as 66 for cell clusters on gold film is a ratio of fluorescent emission peak area measured at the CD labeled cell clusters to the fluorescent peak area measured at unlabeled cell clusters (autofluorescence) on the same substrate. The contrast with the background (S/N) or the ratio of fluorescent peak area measured at bacteria cells to area measured at bare substrate was as high as 200. This report may pave a way for the broader application of surface enhanced fluorescence and especially metal enhanced fluorescence imaging of CD labeled cells and other biological objects.
Graphical abstract Carbon dots, synthesized from dates, are used for direct staining of E coli cells. Emission fluorescent spectroscopy of those CD labelled cells on gold film and glass, demonstrated enhancement factor about 50 for emission on gold as compared to glass, Excitation at 633 nm appears far superior to excitation at 532 nm in terms of contrast (up to 67) with unlabeled cells /control due to decrease in auto fluorescence of cells. Maximum Signal to noise ratio is 200.
The spectral and optical properties of the fractionated components of dissolved organic matter (DOM) of three freshwater lakes in Karelia were studied using reversed-phase high-performance liquid chromatography (RP-HPLC) with online detection of fluorescence and absorption spectra. It is shown that the DOM fractions are qualitatively similar, but differ quantitatively in the ratio of components and consist of at least three types of fluorophores: (1) hydrophilic “humic-like” fluorophore(s) with the emission maximum in the region of 420 nm and an absorption band at 260–270 nm; (2) hydrophobic “humic-like” fluorophore(s) with the emission maximum at approximately 450 nm that has no characteristic absorption maxima in the region from 220 to 400 nm; and (3) a “protein-like” fluorophore with the emission maximum in the region of 340–350 nm, which is typical of proteins and peptides containing tryptophan. 相似文献
Upconversion fluorescence emission of Er3+-doped oxyfluorosilicate glass excited at 975 nm is experimentally investigated. The results reveal that the intense green and red emission, and weak blue emission centered at 525, 543, 655, and 410 nm, respectively. A two-photon upconversion process is assigned to the green and red emission while a three-photon process is responsible for blue upconversion. The possible upconversion mechanisms are discussed based on excited state absorption and energy transfer between excited Er3+ ions. The intense upconversion fluorescence of Er3+-doped lead oxyfluorosilicate glass may be a potentially useful material for developing upconversion optical devices. 相似文献
The aim of this research is to study the normalized fluorescence spectra (intensity variations and area under the fluorescence
signal), relative quantum yield, extinction coefficient and intracellular properties of normal and malignant human bone cells.
Using Laser-Induced Fluorescence Spectroscopy (LIFS) upon excitation of 405 nm, the comparison of emission spectra of bone
cells revealed that fluorescence intensity and the area under the spectra of malignant bone cells was less than that of normal.
In addition, the area ratio and shape factor were changed. We obtained two emission bands in spectra of normal cells centered
at about 486 and 575 nm and for malignant cells about 482 and 586 nm respectively, which are most likely attributed to NADH
and riboflavins. Using fluorescein sodium emission spectrum, the relative quantum yield of bone cells is numerically determined. 相似文献
The study reports synthesis and photophysical studies of a new zinc sensing pyrazole scaffold structurally characterized to be bis(2-hydroxybenzylidene)-1H-pyrazole-3,5-dicarbohydrazide (PHSA). Excitation of the probe at 330 nm results in an emission band at 417 nm which ratiometrically red shifts to 466 nm upon Zn2+ addition in an unprecedented way. The photo induced electron transfer (PET) coupled intramolecular charge transfer (ICT) working for a dual-channel fluorescence emission pathway is observed and studies were supported with density functional theory and NMR titration experiments. The probe exhibited dissociation constant of 1.2156 and detection limit as low as 992 nM. The cytotoxic effects of the probe on 60 tumour cell lines were tested. The intracellular zinc sensing with reversible binding potential is verified with fluorescence microscopy experiment. 相似文献
We examined the emission spectra and steady-state anisotropy of tyrosinate anion fluorescence with one-photon (250–310 nm), two-photon (570–620 nm) and three-photon (750–930 nm) excitation. Similar emission spectra of the neutral (pH 7.2) and anionic (pH 13) forms of N-acetyl-L-tyrosinamide (NATyrA) (pKa 10.6) were observed for all modes of excitation, with the maxima at 302 and 352 nm, respectively. Two-photon excitation (2PE) and three-photon excitation (3PE) spectra of the anionic form were the same as that for one-photon excitation (1PE). In contrast, 2PE spectrum from the neutral form showed ~30-nm shift to shorter wavelengths relative to 1PE spectrum (λmax 275 nm) at two-photon energy (550 nm), the latter being overlapped with 3PE spectrum, both at two-photon energy (550 nm). Two-photon cross-sections for NATyrA anion at 565–580 nm were 10 % of that for N-acetyl-L-tryptophanamide (NATrpA), and increased to 90 % at 610 nm, while for the neutral form of NATyrA decreased from 2 % of that for NATrpA at 570 nm to near zero at 585 nm. Surprisingly, the fundamental anisotropy of NATyrA anion in vitrified solution at ?60 °C was ~0.05 for 2PE at 610 nm as compared to near 0.3 for 1PE at 305 nm, and wavelength-dependence appears to be a basic feature of its anisotropy. In contrast, the 3PE anisotropy at 900 nm was about 0.5, and 3PE and 1PE anisotropy values appear to be related by the cos6θ to cos2θ photoselection factor (approx. 10/6) independently of excitation wavelength. Attention is drawn to the possible effect of tyrosinate anions in proteins on their multi-photon induced fluorescence emission and excitation spectra as well as excitation anisotropy spectra. 相似文献
Ultraviolet upconversion signals at 293, 351, and 366 nm were observed from thulium-doped fluorozirconate fiber pumped with a 458-nm Ar(+) laser. The upconversion signal's intensity was enhanced 5x when the fiber was simultaneously pumped with a 458-nm Ar(+) laser and a 585-nm dye laser. There is evidence of the formation of defect centers under simultaneous excitation by visible and near-infrared lasers (458 and 750 nm), and there is no evidence of color-center formation when both of the exciting beams are in the visible (458 and 585 nm). 相似文献