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At least 91 records · Page 5Linked to original sources
PubMed2002

Macro-microscopic fluorescence imaging of human NPC xenografts in a murine model using topical vs intravenous administration of 5-aminolevulinic acid.

The use of 5-aminolevulinic acid to induce endogenous porphyrins for the purpose of detection of epithelial cancers is being studied extensively in many centres around the world. The challenge is to prepare an efficacious formulation of 5-ALA for the purpose of cancer detection. In this study, we compared two formulations of topical 5-ALA applications with intravenous administration in NPC/CNE-2 xenografts on balb/c nude mice. One of the formulations was a gantrez muco-adhesive patch and the other was a polyvinyl-pyrolidone muco-adhesive patch. The Karl Storz fluorescence endoscopy system was used to obtain macroscopic fluorescence images. Microscopic fluorescence imaging was done by laser confocal microscopy. The macroscopic images were further analysed for fluorescence intensity distribution. It was found that between the two formulations of topical application of 5-ALA; there was very little difference in the fluorescence biodistribution. When the topical applications were compared with the intravenous administration, the tumour to normal differential in biodistribution was significantly higher with the topical application compared to the intravenous application.

Administration, Topical↗
PubMed2003

Rapid, noninvasive screening for perturbations of metabolism and plant growth using chlorophyll fluorescence imaging.

A rapid, noninvasive technique involving imaging of chlorophyll fluorescence parameters for detecting perturbations of leaf metabolism and growth in seedlings is described. Arabidopsis seedlings were grown in 96-well microtitre plates for 4 d and then treated with eight herbicides with differing modes of action to induce perturbations in a range of different metabolic processes. Imaging of chlorophyll fluorescence emissions from 96 seedlings growing on a microtitre plate enabled images of a number of fluorescence parameters to be rapidly and simultaneously produced for the plants in each well. Herbicideinduced perturbations in metabolism, even in metabolic reactions not directly associated with photosynthetic metabolism, were detected from the changes in the images of fluorescence parameters considerably before any visual effects on seedling growth were observed. Evaluations of seedling growth were made from measurements of the area of chlorophyll fluorescence emission in images of plants growing in the 96-well plates. Decreased seedling growth related directly to herbicideinduced changes in the imaged chlorophyll fluorescence parameters. The applicability of this rapid-screening technique for metabolic perturbations in monocotyledonous species was demonstrated by treating Agrostis tenuis seedlings with Imazapyr, an inhibitor of branched-chain amino acid synthesis.

Acetone↗
PubMed2007

Quantitative assessment of small intraosseous prostate cancer burden in SCID mice using fluorescence imaging.

BACKGROUND: Experimental bone metastases are typically analyzed when the skeletal tumor burden is large enough to be detected by imaging or histology. By this time, the bone microenvironment is usually destroyed, preventing useful analysis of tumor-bone interactions. METHODS: Small intraosseous tumors generated by intratibial injection of C4-2B prostate cancer cells transfected with green fluorescent protein (GFP) were assessed using in vivo and ex vivo fluorescence imaging, radiography, histology, and fluorometric analysis of bone lysates. RESULTS: Ex vivo fluorescence imaging and fluorometric analysis were capable of detecting tiny bone tumors as early as 10 days after injection. Ex vivo fluorescence imaging allowed simple quantification of small skeletal tumor burden and was useful in measuring the effect of systemic therapy. CONCLUSIONS: Ex vivo fluorescence imaging is a sensitive and easy method to quantify small skeletal tumor burden. This technique allows investigation of tumor-bone interactions while the bone microanatomy is still intact.

Animals↗
PubMed2007

Quantization of widefield fluorescence images using structured illumination and image analysis software.

It is difficult to obtain precise quantitative measurements from fluorescent images captured from widefield microscopes. We wished to ascertain if reliable quantitative measurements of both biological and nonbiological specimens were possible using a widefield microscope equipped with a structured illumination system and image analysis software. In a nonbiological specimen, images were obtained from fluorescent beads of known intensity. For a biologically relevant model we used the uptake of fluorescently labeled transferrin by HeLa cells in culture. In the bead sample, the mean intensity of beads acquired with the structured illumination system showed relative intensities near to the predicted values without any further manipulation of data. In the transferrin uptake experiments, uptake and subsequent recycling of the labeled transferrin could be accurately and reproducibly measured. We conclude that using a combination of structured illumination and image processing algorithms accurate quantization of fluorescent images can be achieved in widefield microscopy.

Algorithms↗
PubMed2005

Fluorometric and fluorescent image analysis methods for determination of lipid hydroperoxides in oil models with 3-perylene diphenylphosphine (3-PeDPP).

Fluorometric and fluorescent image analysis methods with 3-perylene diphenylphosphine (3-PeDPP) were developed to measure lipid hydroperoxides in refined tuna oil. The fluorescence intensity of 3-perylene diphenylphosphineoxide (3-PeDPPO, the corresponding peroxidation product of 3-PeDPP) has been used for assessing lipid hydroperoxides in refined tuna oil models, and the results are compared to those of a classical method using ferric-xylenol orange complex. There were good correlations between the lipid hydroperoxide contents determined by the 3-PeDPPO fluorescence and the xylenol orange methods (R(2) = 0.984). Moreover, the novel probe 3-PeDPP and the fluorescent image analysis enabled us to evaluate two-dimensionally the lipid hydroperoxide contents in tuna oil droplets. These results suggest that the fluorometric and fluorescent image analyses using 3-PeDPP would be suitable for batch and two-dimensional determinations of lipid hydroperoxides in oil and oil-containing materials.

Animals↗
PubMed2002

Initial neural net construction for the detection of cervical intraepithelial neoplasia by fluorescence imaging.

OBJECTIVE: The aim of this study was to initiate neural net construction for the detection of cervical intraepithelial neoplasia by fluorescence imaging. STUDY DESIGN: Thirty-three women with abnormal Papanicolaou smears underwent fluorescence imaging during colposcopy. With the use of >4000 training pixels and >1000 test pixels, intrapatient nets were constructed from the spectral data of 17 women. An interpatient net that discriminated between cervical intraepithelial neoplasia 1 and normal tissue classes among patients was constructed with the use of >2300 training pixels and >2000 test pixels from 12 women. Average correct classification rates were determined. Sensitivities, specificities, and positive and negative predictive values for cervical intraepithelial neoplasia grade 1 and normal tissue classes were calculated. Extrapolated false-color cervical images were generated. RESULTS: Average correct classification rates were 96.5% for the intrapatient nets and 97.5% for the interpatient net. The sensitivity, specificity, and positive and negative predictive values for cervical intraepithelial neoplasia grade 1 were 98.2%, 98.9%, 71.4%, and 99.9%, respectively. CONCLUSION: Initial results suggest that neural nets that are constructed from fluorescence imaging spectra may offer a potential method for the detection of cervical intraepithelial neoplasia.

Female↗
PubMed2004

Whole-column fluorescence-imaged capillary electrophoresis.

An axially illuminating whole-column fluorescence imaging capillary electrophoresis (CE) experimental setup was developed. A 6 cm long Teflon tube with an inside diameter (ID) of 42 microm was used as separation column. Excitation light of 488 nm from Ar+ laser was introduced to one end of the separation column by an optical fiber. The excitation light propagated inside the separation column by total internal reflection, since the refractive index of the buffer solution was larger than that of the Teflon tube. The fluorescence from the whole separation column was imaged with a charge-coupled device (CCD) camera. Fluorescent compounds such as fluorescein isothiocyanate (FITC), 5-carboxyfluorescein, and FITC-labeled protein were used to test the basic performance of the experimental setup. Experimental results illustrate that the whole-column-fluorescence imaging CE is a fast and sensitive separation method for fluorescent compounds and fluorescent-labeled proteins. Furthermore, it could be used for simple, fast, and easy comparisons of the resistance to photodegradation for various fluorescent compounds.

Electrophoresis, Capillary↗
PubMed1988

Simultaneous Nomarski and fluorescence imaging during video microscopy of cells.

A video microscope designed to allow low light level fluorescence imaging of cells during simultaneous high-resolution differential interference contrast (DIC) imaging, without the fluorescence light losses of 60-90% normally associated with this contrast-enhancement technique, is described. Transmitted light for DIC imaging, filtered at greater than 620 nm, passes through standard DIC optical components, (1/4 lambda-plate, polarizer, and Wollaston prism) before illuminating the cells. Transmitted light and fluorescence emission pass through a second Wollaston prism but not through the analyzer, which is repositioned more distally in the optical path. Prisms designed to reflect light out a side port of the microscope to a video camera have been replaced with a dichroic mirror. This mirror reflects fluorescence emission out the side port to a low light-sensitive video camera. The spectrally distinct transmitted light continues through the dichroic mirror to an overhead camera through a polarizer (analyzer), which completes the DIC optical path. The fluorescence and DIC images can be viewed simultaneously on side-by-side video monitors, examined sequentially by an image-processing computer, or examined simultaneously using a video splitter/inserter. The ability to image cells with high resolution simultaneously with low light level fluorescence imaging should find wide applicability whenever it is necessary or desirable to correlate fluorescence intensity or distribution with specific cell structure or function.

Animals↗
PubMed1999

Detection of premalignant oral lesions in hamsters with an endoscopic fluorescence imaging system.

BACKGROUND: Various methods of detecting cancer with fluorescence have been developed. One type of fluorescence is based on the tumor-localizing properties of certain dyes. However, the phototoxicity of most known tumor-localizing dyes hinders the safe use of such diagnostic methods. The authors have developed a fluorescence imaging system to detect the distribution of a nontoxic dye, fluorescein, and they have evaluated the feasibility of the system by using it to detect oral dysplastic lesions in hamsters. METHODS: Dysplasia was induced in the cheek pouches of hamsters by application of the carcinogen 9,10-dimethyl-1,2-benzantracene. Fluorescein was administered to the hamsters either intravenously or orally before the fluorescence examination. The endoscopic fluorescence system produced dye-distribution images of both treated and control pouches. Two fluorescence images in different spectral regions were processed for each dye image. Biopsy material from both pouches was examined histopathologically. RESULTS: The accumulation of fluorescein was detected in 22 of 23 specimens containing dysplastic lesions. CONCLUSIONS: These results demonstrate the utility of this fluorescein accumulation method in the detection of dysplasia. The accumulation of fluorescein in dysplastic lesions may point to acidification of interstitial medium in such lesions.

Administration, Oral↗
PubMed2000

Microspectrofluorometry and fluorescence imaging in the study of human cytopathology.

The study of energy pools and dynamics of specific pathways in living cells by microspectrofluorometry and fluorescence imaging produces spectral and topographic images characterizing structural and functional changes associated with cytopathology. Microspectro-fluorometry and fluorescence imaging have been applied, together with organelle morphometry to a number of cells mimicking certain cytopathologies, including melanoma cells, long-term malignant cells, and gene-defective cells. These investigations of cellular pathology indicate that there is a convergence of various physiopathological processes. Cellular states that have similarities include senescence, detoxification, and transformation. While the NAD(P)H metabolic transients have been studied before, our emphasis in this article is on very rapidly scanned fluorescence images related to organelle integration and photoinduced cellular senescence.

Animals↗
PubMed2006

Intraoperative assessment of reconstructed vessels in living-donor liver transplantation, using a novel fluorescence imaging technique.

BACKGROUND/PURPOSE: In living-donor liver transplantation (LDLT), hepatic arterial thrombosis and portal venous thrombosis are critical problems that can result in graft loss. Only intraoperative Doppler ultrasound (IDUS) is able to evaluate blood flow in the reconstructed vessels. The aim of this study was to evaluate the utility of a newly developed fluorescence imaging technique using indocyanine green (ICG) for visualizing reconstructed vessels. METHODS: In three patients who had undergone LDLT, IDUS was performed after reconstruction of the portal vein and hepatic artery. Fluorescence images were then recorded, using a SPY system (Novadeq Technologies), which employs ICG as a fluorescent imaging medium activated by light. The ICG (3.75 mg) was injected intravenously, then, 10 s later, the images were recorded for 30 s (first photographic recording). Two minutes later, the same procedure was repeated (second photographic recording), and 40 min later, images were obtained without injection of ICG (third photographic recording). RESULTS: After portal venous reconstruction, IDUS demonstrated a nonphasic and continuous waveform, with a mean velocity of 52.1 cm/s and a mean portal blood flow volume of 69.5 ml/s per kg. After hepatic arterial reconstruction, a pulsatile waveform with a mean peak systolic velocity of 52.4 cm/s and a mean resistance index of 0.76 was obtained. The first photographic recording clearly visualized the blood flow in the reconstructed hepatic artery, without kinking or stenosis, in all three patients. The second photographic recording visualized the flow in the portal vein without stenosis, kinking, or stagnation. The third photographic recording demonstrated the excretion of ICG into bile, thus confirming bile production by the grafts. CONCLUSIONS: Fluorescence imaging can clearly visualize the reconstructed hepatic artery and portal vein and demonstrate the production of bile by a transplanted liver graft. A combination of IDUS and the new system can guarantee the patency of the reconstructed vessels.

Blood Flow Velocity↗
PubMed2004

MR and fluorescent imaging of low-density lipoprotein receptors.

RATIONALE AND OBJECTIVES: Over-expression of low-density lipoprotein receptors (LDLRs) occurs in many types of malignancies and is related to the requirement for lipids for rapid proliferation of the tumors. On the other hand, LDLRs that are unable to bind LDL are found on hepatocytes of patients with familial hypercholesterolemia (FH), a genetic disease that leads to premature atherosclerosis and death. The highly selective binding of LDL to LDLR makes these particles ideal carriers of therapeutic and diagnostic contrast agents into the targeted cells. The objectives of this paper are to examine whether a prototype contrast agent (PTIR267) with dual detection properties is suitable for labeling of LDL particles for in vivo detection of LDLR by magnetic resonance imaging (MRI) and for in vitro monitoring of cellular localization by confocal fluorescence microscopy. MATERIALS AND METHODS: PTIR267 is a lipophilic GdDTPA derivative conjugated to a fluorescent dye. The conjugated dye molecule makes the probe sufficiently water soluble to allow labeling of LDL by a brief incubation of LDL with PTIR267 dissolved in PBS at 37 degrees C (mole ratio LDL: PTIR267 = 0.09:1). The molar relaxivity of PTIR267 in saline is 26 mM(-1)s(-1). Specific LDLR-mediated uptake of PTIR267-labeled LDL was demonstrated in vitro by confocal fluorescence imaging of B16 melanoma cells using confocal fluorescence imaging. In vivo uptake of PTIR267-labeled LDL by a subcutaneously implanted B16 melanoma in mice leads to 30% decrease in longitudinal relaxation time (T(1)) in the tumor. In vivo uptake of PTIR267-labeled LDL leads to 70% decrease in T(1) in a normal C57BL/6 mouse liver; however, in the liver of LDL receptor gene knockout (LDLr-/-) mice with C57BL/6 background, only 12% decrease in T(1) is observed. CONCLUSIONS: The dual fluorescence and MR imaging properties of PTIR267, combined with the ease of LDL labeling, suggest that it will be a useful tool for optimization of LDLR-targeted cancer diagnosis or therapy and for monitoring the efficacy of gene therapy of FH.

Animals↗
PubMed2006

Fluorescence imaging and phototoxicity effects of new formulation of chlorin e6-polyvinylpyrrolidone.

Evaluations of the efficiency of a new formulation of chlorin consisting of a complex of trisodium salt chlorin e6 (Ce6) and polyvinylpyrrolidone (PVP) in photodynamic therapy (PDT) and fluorescence diagnosis was performed on poorly differentiated human bladder carcinoma murine model with the following specific aims: (i) to qualitatively evaluate the fluorescence accumulation in human bladder tumor, (ii) to determine fluorescence distribution of Ce6-PVP using the tissue extraction technique and fluorescence imaging technique, (iii) to compare the fluorescence distribution of Ce6, Ce6-PVP and Photofrin in skin of nude mice, and (iv) to investigate phototoxicity caused by different parameters (drug-light interval, drug dose, irradiation fluence rate and total light fluence) in PDT. The fluorescence of the Ce6-PVP formulation was determined either by fluorescence imaging measurements or by chemical extraction from the tissues displaying similar trends of distribution. Our results demonstrated that the Ce6-PVP formulation possesses less in vivo phototoxic effect compared to Ce6 alone. The phototoxicity revealed a strong dependence on the drug and light dosimetry as well as on the drug-light interval. In PDT, the Ce6-PVP compound was most toxic at the 1h drug-light interval at 200J/cm(2), while Ce6 alone was most toxic at a light dose of more that 50J/cm(2) at the 1 and 3h drug-light interval. We also confirmed that Ce6-PVP has a faster clearance compared to Ce6 alone or Photofrin. This eliminates the need for long-term photosensitivity precautions. In conclusion, the Ce6-PVP formulation seems to be a promising photosensitizer for fluorescence imaging as well as for photodynamic treatment.

Animals↗
PubMed2006

Spatial-temporal variations in rose leaves under water stress conditions studied by chlorophyll fluorescence imaging.

Spatial-temporal changes were examined by imaging chlorophyll (Chl) a fluorescence in four leaf areas, two central and two external of rose plants (Rosa x hybrida) cv. Grand Gala for 9 days, under progressive water stress. New fluorescence parameters based on the lake model have recently been used to determine Q(A) redox state and excitation energy fluxes in order to gain a better understanding of the mechanisms that occur under drought stress. Chlorophyll fluorescence images showed a spatial variation in the leaves. The lower values for F(o), F(M), phi(2), q(P) and q(L) were found in the internal leaf area while higher values of non-photochemical quenching calculated from Stern-Volmer quenching (NPQ) and phi(NPQ). phi(Po) were more homogeneous throughout leaf. Temporal changes were also observed during the experiment, a 10% decrease in relative water content (RWC) (between day 1 and 2), led to a decrease in photochemical quenching and an increase in non-photochemical processes. Chlorophyll fluorescence parameters were more or less constant till day 8. At the end of the experiment (day 9), energy dissipation by downregulation, electron transport and Q(A) redox state, decreased and phi(NO) increased to compensate the change. Chlorophyll fluorescence parameters based on the lake model q(L), phi(NPQ) and phi(NO) have been found more appropriate for estimating the fraction of open centres, the quantum yield of regulated energy dissipation in photosystem II (PSII) and the quantum yield of non-regulated energy dissipation in PSII, respectively. The F(s)/F(o) ratio is strongly correlated with NPQ and phi(NPQ) up to a RWC of 20%. This coincides with a greater decrease in photochemical quenching and non-photochemical quenching and an increase in phi(NO).

Chlorophyll↗
PubMed2001

Simultaneous topographic and fluorescence imaging of single DNA molecules for DNA analysis with a scanning near-field optical/atomic force microscope.

High-resolution fluorescence imaging of lambda-phage DNA molecules, intercalated with the dye YOYO-1, has been performed by a SNOM/AFM based on a bent-type optical fiber probe. A modified design of the optical probe has been made, and successful near-field optical resolution has been obtained for the strongly stretched lambda-phage DNA molecules. The best optical resolution was estimated at 45 nm for the dye-intercalated single lambda-DNA molecules by a mean width evaluation. In our comparison between the far-field fluorescence and high-resolution near-field fluorescence images for the DNA, it has been found that the near-field images much better defined the intercalation state of the dye. Finally, the relation between the DNA shapes and the dye distribution states, and the discrimination between the double-stranded and single-stranded DNA molecules, are discussed by comparing the topography and fluorescence images of the SNOM/AFM.

Benzoxazoles↗
PubMed2002

Platelets, circulating tissue factor, and fibrin colocalize in ex vivo thrombi: real-time fluorescence images of thrombus formation and propagation under defined flow conditions.

Although it is generally accepted that the initial event in coagulation and intravascular thrombus formation is the exposure of tissue factor (TF) to blood, there is still little agreement about the mechanisms of thrombus propagation and the identities of the molecular species participating in this process. In this study, we characterized the thrombotic process in real-time and under defined flow conditions to determine the relative contribution and spatial distribution of 3 components of the thrombi: circulating or blood-borne TF (cTF), fibrin, and platelets. For this purpose, we used high-sensitivity, multicolor immunofluorescence microscopy coupled with a laminar flow chamber. Freshly drawn blood, labeled with mepacrine (marker for platelets and white cells), anti-hTF1(Alexa.568) (marker for tissue factor), and anti-T(2)G(Cy-5)(1) (marker for fibrin) was perfused over collagen-coated glass slides at wall shear rates of 100 and 650 s(-1). A motorized filter cube selector facilitated imaging every 5 seconds at 1 of 3 different wavelengths, corresponding to optimal wavelengths for the 3 markers above. Real-time video recordings obtained during each of 10 discrete experiments show rapid deposition of platelets and fibrin onto collagen-coated glass. Overlay images of fluorescent markers corresponding to platelets, fibrin, and cTF clearly demonstrate colocalization of these 3 components in growing thrombi. These data further support our earlier observations that, in addition to TF present in the vessel wall, there is a pool of TF in circulating blood that contributes to the propagation of thrombosis at a site of vascular injury.

Antibodies↗
PubMedDate not supplied

Confocal fluorescence imaging of photosensitized DNA denaturation in cell nuclei.

The double-stranded helical structure of DNA is maintained in part by hydrogen bonds between strands and by stacking interactions between adjacent purine and pyrimidine bases in one strand. The transition (denaturation) from a double-stranded (ds) to a single-stranded (ss) form can be induced in isolated DNA or fixed cells by exposure to elevated temperatures, alkali or acids, aprotic or nonpolar solvents or some drugs. We report here that DNA denaturation can occur in situ in cell nuclei as a result of interaction between light and an intercalated dye, acridine orange or ethidium bromide. This DNA photodenaturation was probed using metachromatic properties of acridine orange and imaged by fluorescence confocal microscopy. Furthermore, an empirical kinetic model was developed to separate changes of acridine orange luminescence intensities caused by photobleaching from those that were a result of DNA denaturation. We investigated the influence of oxygen on these phenomena and propose a mechanism by which photodenaturation may occur.

Cell Line↗