Home About us Contact
|
Home About us Contact | ||
|
|
||
Fluorescence Images (fluorescence + image)
Selected AbstractsCover Picture: J. Biophoton.JOURNAL OF BIOPHOTONICS, Issue 4 20094/200 AC Dielectrophoretic separation of 10 ,m non-fluorescent microspheres from 40 nm red fluorescent nanoparticles on a microelectrode array. The leftmost 3 × 3 microelectrodes are activated. The rightmost column of microelectrodes are used as control. (Top Left) Red Fluorescence image before experiment showing a red haze. (Top Right) Red Fluorescence image of the 40 nm nanoparticles concentrating on the electrodes at the conclusion of the experiment. (Bottom Left) Bright Field image of 10 ,m microspheres distributed in a random pattern before the experiment. (Bottom Right) Bright Field image of 10 ,m microspheres concentrating in between the microelectrodes at the conclusion of the experiment (© 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Selective detection of superoxide anion radicals generated from macrophages by using a novel fluorescent probeFEBS JOURNAL, Issue 7 2007Jing Jing Gao Quantitation of superoxide radical (O,2,·) production at the site of radical generation remains challenging. A simple method to detect nanomolar to micromolar levels of superoxide radical in aqueous solution has been developed and optimized. This method is based on the efficient trapping of O2,· using a novel fluorescent probe (2-chloro-1,3-dibenzothiazolinecyclohexene), coupled with a spectra character-signaling increase event. A high-specificity and high-sensitivity fluorescent probe was synthesized in-house and used to image O2,· in living cells. Better selectivity for O2,· over competing cellular reactive oxygen species and some biological compounds illustrates the advantages of our method. Under optimal conditions, the linear calibration range for superoxide anion radicals was 5.03 × 10,9,3.33 × 10,6 m. The detection limit was 1.68 × 10,9 m. Fluorescence images of probe-stained macrophages stimulated with 4,-phorbol 12-myristate 13-acetate were obtained successfully using a confocal laser scanning microscope. [source] Affinity-Based Protein Surface Pattern Formation by Ligand Self-Selection from Mixed Protein SolutionsADVANCED FUNCTIONAL MATERIALS, Issue 19 2009Manish Dubey Abstract Photolithographically prepared surface patterns of two affinity ligands (biotin and chloroalkane) specific for two proteins (streptavidin and HaloTag, respectively) are used to spontaneously form high-fidelity surface patterns of the two proteins from their mixed solution. High affinity protein-surface self-selection onto patterned ligands on surfaces exhibiting low non-specific adsorption rapidly yields the patterned protein surfaces. Fluorescence images after protein immobilization show high specificity of the target proteins to their respective surface patterned ligands. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging further supports the chemical specificity of streptavidin and HaloTag for their surface patterned ligands from mixed protein solutions. However, ToF-SIMS did detect some non-specific adsorption of bovine serum albumin, a masking protein present in excess in the adsorbing solutions, on the patterned surfaces. Protein amino acid composition, surface coverage, density, and orientation are important parameters that determine the relative ToF-SIMS fragmentation pattern yields. ToF-SIMS amino acid-derived ion fragment yields summed to produce surface images can reliably determine which patterned surface regions contain bound proteins, but do not readily discriminate between different co-planar protein regions. Principal component analysis (PCA) of these ToF-SIMS data, however, improves discrimination of ions specific to each protein, facilitating surface protein pattern identification and image contrast. [source] Delineating melanoma using multimodal polarized light imagingLASERS IN SURGERY AND MEDICINE, Issue 1 2009Zeina Tannous Abstract Background and Significance Melanoma accounts for 3% of all skin cancers but causes 83% of skin cancer deaths. The first step in treatment of melanoma is the removal of the lesions, usually by surgical excision. Currently most lesions are removed without intraoperative margin control. Post-operative methods inspect 1,2% of the surgical margin and are prone to sampling errors. In this study we evaluate the use of reflectance and fluorescence polarization imaging for the demarcation of melanoma in thick fresh skin excisions. Materials and Methods Pigmented lesions clinically suspicious for melanoma were elliptically excised with proper margins. Elliptical surgical excisions were vertically bisected along the short axis of the specimen into two halves in the middle of the pigmented lesions. The vertically bisected tumor face was imaged. After that, one half of the sample was briefly stained in aqueous 2 mg/ml solution of tetracycline, whereas another half was stained in 0.2 mg/ml aqueous solution of methylene blue. Then both specimens were reimaged. Reflectance images were acquired in the spectral range between 390 and 750 nm. Fluorescence images of the tetracycline-stained tissue were excited at 390 nm and registered between 450 and 700 nm. Fluorescence of the methylene blue-stained samples was excited at 630 nm and registered between 650 and 750 nm. After imaging, the tissue was processed for standard H&E histopathology. The resulting histological and optical images were compared to each other. Results and Conclusions Our findings demonstrate that both tetracycline and methylene blue are suitable for imaging dysplastic and benign nevi. Melanoma is better delineated in the samples stained in methylene blue. Accurate and rapid delineation of melanoma in standard fresh surgical excisions appears feasible. Lasers Surg. Med. 41:10,16, 2009. © 2008 Wiley-Liss, Inc. [source] Length-Dependent Uptake of DNA-Wrapped Single-Walled Carbon Nanotubes,ADVANCED MATERIALS, Issue 7 2007L. Becker A length threshold for cell uptake of DNA-wrapped single-walled carbon nanotubes (SWNTs) by human lung fibroblasts (IMR90) is identified. Competitive uptake experiments with well-defined and characterized length fractions show that SWNTs above the length threshold are excluded from the cell, whereas SWNTs labeled with Cy3-derivatized DNA below the threshold are able to access the cell interior, as shown in the fluorescence image and on the cover. [source] Analysis of fluorescence from algae fossils of the Neoproterozoic Doushantuo formation of China by confocal laser scanning microscopeMICROSCOPY RESEARCH AND TECHNIQUE, Issue 4 2006Huimei Chi Abstract Chinese algae fossils can provide unique information about the evolution of the early life. Thin sections of Neoproterozoic algae fossils, from Guizhou, China, were studied by confocal laser scanning microscopy, and algae fossils were fluorescenced at different wavelengths when excited by laser light of 488 nm, 476 nm, and 568 nm wavelength. When illuminated by 488 nm laser light, images of the algae fossils were sharper and better defined than when illuminated by 476 nm and 568 nm laser light. The algae fossils fluoresce at a wide range of emission wavelengths. The three-dimensional images of the fluorescent algae fossils were compared with the transmission images taken by light microscope. We found that the fluorescence image of the confocal laser scanning microscope in a single optical section could pass for the transmission image taken by a light microscope. We collected images at different sample depths and made a three-dimensional reconstruction of the algae fossils. And on the basis of the reconstruction of the three-dimensional fluorescent images, we conclude that the two algae fossils in our present study are red algae. Microsc. Res. Tech. 69:253,259, 2006. © 2006 Wiley-Liss, Inc. [source] A Two-Photon Fluorescent Probe for Lipid Raft Imaging: C-LaurdanCHEMBIOCHEM, Issue 5 2007Hwan Myung Kim Abstract The lipid-rafts hypothesis proposes that naturally occurring lipid aggregates exist in the plane of membrane that are involved in signal transduction, protein sorting, and membrane transport. To understand their roles in cell biology, a direct visualization of such domains in living cells is essential. For this purpose, 6-dodecanoyl-2-(dimethylamino)naphthalene (laurdan), a membrane probe that is sensitive to the polarity of the membrane, has often been used. We have synthesized and characterized 6-dodecanoyl-2-[N -methyl- N -(carboxymethyl)amino]naphthalene (C-laurdan), which has the advantages of greater sensitivity to the membrane polarity, a brighter two-photon fluorescence image, and reflecting the cell environment more accurately than laurdan. Lipid rafts can be visualized by two-photon microscopy by using C-laurdan as a probe. Our results show that the lipid rafts cover 38,% of the cell surface. [source] Electrochemical Modulation of Remote Fluorescence Imaging at an Ordered Opto-electrochemical Nanoaperture ArrayCHEMPHYSCHEM, Issue 8 2004Arnaud Chovin Abstract An array of nanometer-sized apertures capable of electrochemically modulating the fluorescence of a model analyte is presented. The device, which combines near-field optical methods and ultramicroelectrode properties in an array format, is based on an etched coherent optical fiber bundle. Indeed, the fabrication steps produced an ordered array where each optical nanoaperture is surrounded by a ring-shaped gold nanoelectrode. The chronoamperometric behavior of the array shows stable diffusion-limited quasi-steady-state response. The model analyte, tris(2,2,-bipyridine) ruthenium, emits fluorescence in the Ru(II) state, but not in the oxidized Ru(III) state. Fluorescence is excited by visible light exiting from each nanoaperture since light is confined to the tip apex by the gold coating. A fraction of the isotropically emitted luminescence is collected by the same nanoaperture, transmitted by the corresponding fiber core and eventually detected by a charge-coupled device (CCD) camera. The array format provides a fluorescence image resolved at the nanometric scale which covers a large micrometric area. Therefore the high-density array plays a bridging role between these two fundamental scales. We established that the opto-electrochemical nanoapertures are optically independent. Fluorescence of the sample collected by each nanoaperture is modulated by changing the potential of the nanoring electrodes. Reversible electrochemical switching of remote fluorescence imaging is performed through the opto-electrochemical nanoaperture array itself. Eventually this ordered structure of nanometer light sources which are electrochemically manipulated provides promising photonic or electro-optical devices for various future applications. For example, such an array has potential in the development of a combined SNOM-electrochemical nanoprobe array to image a real sample concomitantly at the nanometer and micrometer scale. [source] Correlation of fluorescence and electron microscopy of F-actin-containing sensory cells in the epidermis of Convoluta pulchra (Platyhelminthes: Acoela)ACTA ZOOLOGICA, Issue 1 2002R Pfistermüller Abstract Phalloidin-stained whole mounts of acoel turbellarians show brightly fluorescing club-shaped structures distributed over the epidermis and concentrated especially at the anterior and posterior tips of the body. By correlating electron micrographic images and fluorescence images of Convoluta pulchra, these structures can be seen to be sensory receptors with a central cilium surrounded by a collar of microvilli. The other candidate for showing fluorescence in the epidermis, namely gland necks, can be ruled out since their distribution is too dense to resemble the distribution of the fluorescent structures seen here. The collared sensory receptors were inserted between epidermal cells, and each bore a central cilium surrounded by a collar of 6,18 microvilli and an additional centrally positioned 2,7 microvilli of which 2 or 3 were associated with a modified rootlet called the swallow's nest. Confocal scanning laser microscopy resolved the core of actin filaments within the microvilli of the collar and their rootlet-like connections to the base of the sensory cell. Such receptors could also be identified by fluorescence microscopy in several other species of acoel turbellarians. [source] Synchrony of spontaneous calcium activity in mouse neocortex before synaptogenesisEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 4 2007Jean-Claude Platel Abstract Spontaneous calcium activity can be detected in embryonic mouse cortical slices as fluorescence intensity variations, in the presence of a fluorescent calcium indicator. Current methods to detect and quantify these variations depend heavily on experimenters whose judgement may interfere with measurement. In the present work, we developed new software called CalSignal for automatic detection and tracking of cellular bodies and quantification of spontaneous calcium activity on time-series of confocal fluorescence images. Analysis of 28 neocortical slices revealed that 21.0% of detected cells displayed peaks of fluorescence corresponding to spontaneous activity, with a mean frequency of one peak per 4 min. This activity was blocked in the absence of extracellular calcium but was not modified after depletion of calcium stores with thapsigargin or blockade of voltage-gated calcium channels with Ni2+. Further, statistical analysis of calcium activity revealed concomitant activation of distant cells in 24 slices, and the existence of a significant network of synchrony based on such coactivations in 17 slices out of 28. These networks enclosed 84.3% of active cells, scattered throughout the neocortical wall (mean distance between cellular bodies, 111.7 µm). Finally, it was possible to identify specific cells which were synchronously active with more neighbouring cells than others. The identity of these nodal cells remains to be investigated to fully comprehend the role of spontaneous calcium activity, before synaptogenesis, in shaping cortical neurogenesis. [source] Cover Picture: J. Biophoton.JOURNAL OF BIOPHOTONICS, Issue 10 200910/200 Transfection of actin,antibody nanodiamond immunoconjugates into HeLa cells using dendrimers (a,c), protamine sulfate (d,f) and cationic liposomes (g,i). Left column: bright-field images; middle column: fluorescence images; right column: overlay images. (Picture: M. Mkandawire et al., see also pp. 596,606, in this issue) [source] Weighting hyperspectral image data for improved multivariate curve resolution resultsJOURNAL OF CHEMOMETRICS, Issue 9 2008Howland D. T. Jones Abstract The combination of hyperspectral confocal fluorescence microscopy and multivariate curve resolution (MCR) provides an ideal system for improved quantitative imaging when multiple fluorophores are present. However, the presence of multiple noise sources limits the ability of MCR to accurately extract pure-component spectra when there is high spectral and/or spatial overlap between multiple fluorophores. Previously, MCR results were improved by weighting the spectral images for Poisson-distributed noise, but additional noise sources are often present. We have identified and quantified all the major noise sources in hyperspectral fluorescence images. Two primary noise sources were found: Poisson-distributed noise and detector-read noise. We present methods to quantify detector-read noise variance and to empirically determine the electron multiplying CCD (EMCCD) gain factor required to compute the Poisson noise variance. We have found that properly weighting spectral image data to account for both noise sources improved MCR accuracy. In this paper, we demonstrate three weighting schemes applied to a real hyperspectral corn leaf image and to simulated data based upon this same image. MCR applied to both real and simulated hyperspectral images weighted to compensate for the two major noise sources greatly improved the extracted pure emission spectra and their concentrations relative to MCR with either unweighted or Poisson-only weighted data. Thus, properly identifying and accounting for the major noise sources in hyperspectral images can serve to improve the MCR results. These methods are very general and can be applied to the multivariate analysis of spectral images whenever CCD or EMCCD detectors are used. Copyright © 2008 John Wiley & Sons, Ltd. [source] Digital photography: A primer for pathologistsJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 2 2004Roger S. Riley Abstract The computer and the digital camera provide a unique means for improving hematology education, research, and patient service. High quality photographic images of gross specimens can be rapidly and conveniently acquired with a high-resolution digital camera, and specialized digital cameras have been developed for photomicroscopy. Digital cameras utilize charge-coupled devices (CCD) or Complementary Metal Oxide Semiconductor (CMOS) image sensors to measure light energy and additional circuitry to convert the measured information into a digital signal. Since digital cameras do not utilize photographic film, images are immediately available for incorporation into web sites or digital publications, printing, transfer to other individuals by email, or other applications. Several excellent digital still cameras are now available for less than $2,500 that capture high quality images comprised of more than 6 megapixels. These images are essentially indistinguishable from conventional film images when viewed on a quality color monitor or printed on a quality color or black and white printer at sizes up to 11×14 inches. Several recent dedicated digital photomicroscopy cameras provide an ultrahigh quality image output of more than 12 megapixels and have low noise circuit designs permitting the direct capture of darkfield and fluorescence images. There are many applications of digital images of pathologic specimens. Since pathology is a visual science, the inclusion of quality digital images into lectures, teaching handouts, and electronic documents is essential. A few institutions have gone beyond the basic application of digital images to developing large electronic hematology atlases, animated, audio-enhanced learning experiences, multidisciplinary Internet conferences, and other innovative applications. Digital images of single microscopic fields (single frame images) are the most widely utilized in hematology education at this time, but single images of many adjacent microscopic fields can be stitched together to prepare "zoomable" panoramas that encompass a large part of a microscope slide and closely simulate observation through a real microscope. With further advances in computer speed and Internet streaming technology, the virtual microscope could easily replace the real microscope in pathology education. Later in this decade, interactive immersive computer experiences may completely revolutionize hematology education and make the conventional lecture and laboratory format obsolete. Patient care is enhanced by the transmission of digital images to other individuals for consultation and education, and by the inclusion of these images in patient care documents. In research laboratories, digital cameras are widely used to document experimental results and to obtain experimental data. J. Clin. Lab. Anal. 18:91,128, 2004. © 2004 Wiley-Liss, Inc. [source] Encapsulation and/or Release Behavior of Bovine Serum Albumin within and from Polylactide-Grafted Dextran MicrospheresMACROMOLECULAR BIOSCIENCE, Issue 4 2004Tatsuro Ouchi Abstract Summary: Polylactide (PLA)-grafted dextran (Dex- graft -PLA) of various contents of sugar units was synthesized by anionic polymerization of L -lactide (L -LA) using the alkoxide of partially trimethylsilylated dextran (TMSDex) and subsequently removing the trimethylsilyl (TMS) groups. The copolymer showed different solubility from L -LA homopolymer with increasing the content of sugar units. We prepared bovine serum albumin (BSA)-loaded microspheres (MS)s according to a water-in-oil-in-water emulsion-solvent evaporation/extraction method using methylene chloride/DMSO as an organic cosolvent. MSs prepared from Dex- graft -PLA [MS(Dex- graft -PLA)s] exhibited higher loading efficiency of BSA than MSs prepared from PLLA [MS(PLLA)s]. The in vitro release rate of BSA from MS(Dex- graft -PLA) was faster than that from MS(PLLA). BSA released from MS(Dex- graft -PLA) maintained the secondary structure of native BSA to a great extent, compared with BSA released from MS(PLLA). Confocal fluorescence images of the differential interference micrographs over the fluorescence images of MS(PLLA) and MS(Dex- graft -PLA). [source] Automatic analysis of aqueous specimens for phytoplankton structure recognition and population estimationMICROSCOPY RESEARCH AND TECHNIQUE, Issue 9 2006Karsten Rodenacker Abstract An automatic microscope image acquisition, evaluation, and recognition system was developed for the analysis of Utermöhl plankton chambers in terms of taxonomic algae recognition. The system called PLASA (Plankton Structure Analysis) comprises (1) fully automatic archiving (optical fixation) of aqueous specimens as digital bright field and fluorescence images, (2) phytoplankton analysis and recognition, and (3) training facilities for new taxa. It enables characterization of aqueous specimens by their populations. The system is described in detail with emphasis on image analytical aspects. Plankton chambers are scanned by sizable grids, divers objective(s), and up to four fluorescence spectral bands. Acquisition positions are focused and digitized by a TV camera and archived on disk. The image data sets are evaluated by a large set of quantitative features. Automatic classifications for a number of organisms are developed and embedded in the program. Interactive programs for the design of training sets were additionally implemented. A long-term sampling period of 23 weeks from two ponds at two different locations each was performed to generate a reliable data set for training and testing purposes. These data were used to present this system's results for phytoplankton structure characterization. PLASA represents an automatic system, comprising all steps from specimen processing to algae identification up to species level and quantification. Microsc. Res. Tech., 2006. © 2006 Wiley-Liss, Inc. [source] Photosynthesis within isobilateral Eucalyptus pauciflora leavesNEW PHYTOLOGIST, Issue 4 2006John R. Evans Summary ,,Adult Eucalyptus pauciflora leaves are vertically displayed. They have multiple palisade cell layers beneath both surfaces, interrupted by numerous oil glands. Here, we characterized light absorption, chlorophyll, photosynthetic capacity and CO2 fixation profiles through these leaves. ,,Multiple chlorophyll fluorescence images of leaves viewed in cross-section were made by applying light from different directions. 14CO2 labelling, followed by paradermal cryosectioning, was used to measure profiles of photosynthesis. ,,Photosynthetic capacity peaked 75 µm into the mesophyll beneath each surface and was lowest in the centre of the 600-µm-thick leaf. Predictions by a multilayer model using Beer's law matched the observed profiles of 14C fixation. When constrained to the horizontal, a vertically acclimated leaf gains only 79% of the daily photosynthesis achieved by a horizontally acclimated leaf. However, it outperforms the horizontally acclimated leaf when both are oriented vertically. ,,Each half of the observed profile of photosynthetic capacity closely matches the profile of light absorption through the leaf with unilateral illumination to that surface. Derivation of biochemical parameters from gas exchange measured under unilateral illumination would underestimate the real photosynthetic capacity of these leaves by 21%. [source] Protoporphyrin IX Fluorescence Kinetics and Localization after Topical Application of ALA Pentyl Ester and ALA on Hairless Mouse Skin with UVB-Induced Early Skin CancerPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 3 2000Johanna T. H. M. van den Akker ABSTRACT In order to improve the efficacy of 5-aminolevulinic acid-based (ALA) photodynamic therapy (PDT), different ALA derivatives are presently being investigated. ALA esters are more lipophilic and therefore may have better skin penetration properties than ALA, possibly resulting in enhanced protoporphyrin IX (PpIX) production. In previous studies it was shown that ALA pentyl ester (ALAPE) does considerably enhance the PpIX production in cells in vitro compared with ALA. We investigated the in vivo PpIX fluorescence kinetics after application of ALA and ALAPE to hairless mice with and without UVB-induced early skin cancer. ALA and ALAPE (20% wt/wt) were applied topically to the mouse skin and after 30 min, the solvent was wiped off and PpIX fluorescence was followed in time with in vivo fluorescence spectroscopy and imaging. At 6 and 12 h after the 30 min application, skin samples of visible lesions and adjacent altered skin (UVB-exposed mouse skin) and normal mouse skin were collected for fluorescence microscopy. From each sample, frozen sections were made and phase contrast images and fluorescence images were recorded. The in vivo fluorescence kinetics showed that ALAPE induced more PpIX in visible lesions and altered skin of the UVB-exposed mouse skin, but not in the normal mouse skin. In the microscopic fluorescence images, higher ALAPE-induced PpIX levels were measured in the stratum corneum, but not in the dysplastic layer of the epidermis. In deeper layers of the skin, PpIX levels were the same after ALA and ALAPE application. In conclusion, ALAPE does induce higher PpIX fluorescence levels in vivo in our early skin cancer model, but these higher PpIX levels are not located in the dysplastic layer of the epidermis. [source] Bispectral fluorescence imaging of aggressive basal cell carcinoma combined with histopathological mapping: a preliminary study indicating a possible adjunct to Mohs micrographic surgeryBRITISH JOURNAL OF DERMATOLOGY, Issue 2 2006B. Stenquist Summary Background, Fluorescence imaging is an attractive diagnostic technique for skin tumour demarcation with potential to move to clinical use. Bispectral fluorescence imaging combines skin autofluorescence with ,-aminolaevulinic acid-induced fluorescence. To evaluate the technique, fluorescence data must be compared with the histopathological extent of the tumour, which is the purpose of the current study. Objectives, To investigate the agreement between bispectral fluorescence images and the histopathological tumour boundary of ill-defined basal cell carcinomas (BCCs). After fluorescence imaging the tumours were removed using Mohs micrographic surgery (MMS) to obtain histopathological maps of the tumour boundaries. Methods, Twelve patients with aggressive BCC of mean diameter 16 mm (range 5,32) in the face were included in the study. The patients were subjected to bispectral fluorescence imaging within the 2 months prior to MMS. The fluorescence images and histopathological maps were aligned using image warping. Results, Five patients (42%) showed good agreement with the histopathological mapping and the remaining seven patients (58%) showed partial agreement. Bispectral investigation combining autofluorescence with protoporphyrin IX (PpIX) fluorescence generally yielded better agreement with the histopathological boundaries of the tumours compared with using only the PpIX fluorescence. Conclusions, In this preliminary study the fluorescence has been compared with the histopathological tumour boundaries. The result implies that the technique can be applied as a useful tool for indicating tumour boundary of aggressive BCCs. Further refinement is needed to be able to indicate the exact tumour border. [source] | |||||||||||||||||||