Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2006
To evaluate CD36 expression with quantum dots 605 (QDs 605) on untreated and 7-ketocholesterol (7... more To evaluate CD36 expression with quantum dots 605 (QDs 605) on untreated and 7-ketocholesterol (7KC)-treated monocytic U937 cells by flow cytometry (FCM) and confocal and multiphoton laser scanning microscopy (CLSM). Cells were analyzed by CLSM, following flow cytometric quantification of CD36 expression and 7KC uptake. Image sequences were obtained by spectral analysis in monophoton and multiphoton CLSM and analyzed by the factor analysis of medical image sequences (FAMIS) algorithm to differentiate emission spectra. In CLSM analysis, cell deposits were screened in ultraviolet excitation modes to optimize the possibilities of QDs 605 and have the benefit of nuclei counterstaining by DAPI. FCM and CLSM reveal the expression of CD36 by means of QDs 605. FCM provides information on 7KC uptake. CLSM provides the localization of 7KC vs. DAPI. As factor curves and images show the red, narrow emission of QDs 605 vs. violet and blue emissions of 7KC and DAPI, respectively, a reliable ident...
To evaluate the cytotoxicity of iron nanoparticles on cardiac cells and to determine whether they... more To evaluate the cytotoxicity of iron nanoparticles on cardiac cells and to determine whether they can modulate the biological activity of 7-ketocholesterol (7KC) involved in the development of cardiovascular diseases. Nanoparticles of iron labeled with Texas Red are introduced in cultures of nonbeating mouse cardiac cells (HL1-NB) with or without 7-ketocholesterol 7KC, and their ability to induce cell death, pro-inflammatory and oxidative effects are analyzed simultaneously. Flow cytometry (FCM), confocal laser scanning microscopy (CLSM), and subsequent factor analysis image processing (FAMIS) are used to characterize the action of iron nanoparticles and to define their cytotoxicity which is evaluated by enhanced permeability to SYTOX Green, and release of lactate deshydrogenase (LDH). Pro-inflammatory effects are estimated by ELISA in order to quantify IL-8 and MCP-1 secretions. Pro-oxidative effects are measured with hydroethydine (HE). Iron Texas Red nanoparticles accumulate at t...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2006
To demonstrate CD36 expression with quantum dots (QDs) 525 and/or 605 on human monocytic U937 cel... more To demonstrate CD36 expression with quantum dots (QDs) 525 and/or 605 on human monocytic U937 cells and atherosclerotic tissue sections by means of flow cytometry (FCM) and/or confocal laser scanning microscopy (CLSM). U937 cells and tissue sections were analyzed by means of FCM and/or CLSM. FCM was performed, using different ultraviolet (UV) and visible (488/532 nm) excitation modes. In the visible mode, fluorescence intensities of QDs, phycoerythrin (PE) and fluorescein isothiocyanate (FITC) were compared. Three-dimensional (3-D) sequences of images were obtained by spectral analysis in a CLSM and analyzed by the factor analysis of medical image sequences (FAMIS) algorithm, providing factor curves and images. Factor images are the result of the FAMIS image processing method, which differentiates emission spectra from 3D sequences of images. In CLSM analysis, preparations are screened in a UV excitation mode to optimize the possibilities of QDs and have the benefit of 4',6-diam...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2009
To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse ... more To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse cardiac cells (HL1-NB) cultured without or with 7-ketocholesterol (7KC) found at an increased level in the plasma of atherosclerotic patients and to simultaneously analyze their cytotoxic, proinflammatory and oxidative properties. Flow cytometry (FCM), confocal laser scanning microscopy and subsequent factor analysis image processing were used to characterize the uptake of nanoparticles and to define their cytotoxicity, evaluated by enhanced permeability to SYTOX Green, release of lactate dehydrogenase (LDH) and morphologic nuclear changes determined with Hoechst 33342. Proinflammatory effects were estimated by enzyme linked immunoassay to quantify IL-8 and MCP-1 secretion. The overproduction of reactive oxygen species (ROS) was determined by FCM with hydroethidine. Whereas the nanoparticles had no cytotoxic or inflammatory effects, they could stimulate ROS production. QDs were not incor...
Annual International Conference of the IEEE Engineering in Medicine and Biology Society, 1996
The aim of the authors' study was to provide an efficient computer automated method for real ... more The aim of the authors' study was to provide an efficient computer automated method for real time determination of the passive properties of voltage clamped cells with the smallest possible stimulation amplitude. The method had to be able to determine a wide range of time constants. The authors applied the correlation technique with a binary noise generator. This technique provides
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2009
To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse ... more To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse cardiac cells (HL1-NB) cultured without or with 7-ketocholesterol (7KC) found at an increased level in the plasma of atherosclerotic patients and to simultaneously analyze their cytotoxic, proinflammatory and oxidative properties. Flow cytometry (FCM), confocal laser scanning microscopy and subsequent factor analysis image processing were used to characterize the uptake of nanoparticles and to define their cytotoxicity, evaluated by enhanced permeability to SYTOX Green, release of lactate dehydrogenase (LDH) and morphologic nuclear changes determined with Hoechst 33342. Proinflammatory effects were estimated by enzyme linked immunoassay to quantify IL-8 and MCP-1 secretion. The overproduction of reactive oxygen species (ROS) was determined by FCM with hydroethidine. Whereas the nanoparticles had no cytotoxic or inflammatory effects, they could stimulate ROS production. QDs were not incor...
(i) We studied the effects of a new cromakalim analogue, SR47063, in guinea-pig ventricular cells... more (i) We studied the effects of a new cromakalim analogue, SR47063, in guinea-pig ventricular cells. The experiments were carried out in whole-cell patch clamp with internal and external solutions supposedly similar to the physiological ones. (ii) SR47063 reversibly activated a time-independent current reversing near the potassium equilibrium potential, and a time-dependent current reversing at a more positive potential. Both currents were blocked by application of glibenclamide. (iii) The time-independent and the time-dependent currents were activating for the same concentration of agonist in every cell, this concentration being very different from cell to cell. (iv) The amplitude of the time-dependent current was shown to depend directly neither on agonist concentration nor on potential, but rather on the amplitude of the current flowing during the prepulse before the test pulse. (v) We conclude that SR47063 is a potent KATP channel opener acting at concentrations lower than one micromolar, and that the time-dependent current is likely due to accumulation and depletion of potassium in restricted areas of the cells.
... In Fig. 2(B), the currents for negative voltage steps appear much larger, and time dependent ... more ... In Fig. 2(B), the currents for negative voltage steps appear much larger, and time dependent as earlier9 Note the total A B: 5ms :L 9 IlnA \ 9 J, OOmV: -_ ~-~_~-- __-----__. - , C ImnA D 40 80 Emv = / ~ -1 ~2-to -2 c o -3 E i-4 ~.1 -4o -~0 EmV Fig. ...
Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2007
The aim of this work was to study, in vitro, cell colonization of two biomaterials currently used... more The aim of this work was to study, in vitro, cell colonization of two biomaterials currently used for bone and cartilage repair, this step being important to understand the function of engineered tissues. Current methods that use histological approaches are not always suited to tissue-engineering analysis. We, therefore, set up a protocol to assess cell distribution, utilizing noninvasive confocal microscopy and fluorescent labels with a far red emission wavelength to optimize scaffold transparency and minimize light scattering. Hard (ceramic substitute) and soft (collagen sponge) biomaterials were seeded respectively, on one side of the scaffold, with human fibroblasts and bovine chondrocytes labelled with carbocyanine dyes (DiD and DiR). The mean penetration depth for DiR labelled fibroblasts and chondrocytes in the two scaffolds, around 270 m, was greater than for DiD (136-218 microm) labelled cells. These depths were independent of cell origin but were influenced by the nature of the scaffolds. Collagen sponge is transparent in contrast to ceramic substitutes where measurements could only be made in opened macropores. Besides the limits of the equipment, the limits of the supports were diffusion for collagen sponges and transmission for ceramic substitutes. Confocal microscopy techniques could thus be used to address the question of cell colonization of porous biomaterials in a noninvasive manner.
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2006
To evaluate CD36 expression with quantum dots 605 (QDs 605) on untreated and 7-ketocholesterol (7... more To evaluate CD36 expression with quantum dots 605 (QDs 605) on untreated and 7-ketocholesterol (7KC)-treated monocytic U937 cells by flow cytometry (FCM) and confocal and multiphoton laser scanning microscopy (CLSM). Cells were analyzed by CLSM, following flow cytometric quantification of CD36 expression and 7KC uptake. Image sequences were obtained by spectral analysis in monophoton and multiphoton CLSM and analyzed by the factor analysis of medical image sequences (FAMIS) algorithm to differentiate emission spectra. In CLSM analysis, cell deposits were screened in ultraviolet excitation modes to optimize the possibilities of QDs 605 and have the benefit of nuclei counterstaining by DAPI. FCM and CLSM reveal the expression of CD36 by means of QDs 605. FCM provides information on 7KC uptake. CLSM provides the localization of 7KC vs. DAPI. As factor curves and images show the red, narrow emission of QDs 605 vs. violet and blue emissions of 7KC and DAPI, respectively, a reliable ident...
To evaluate the cytotoxicity of iron nanoparticles on cardiac cells and to determine whether they... more To evaluate the cytotoxicity of iron nanoparticles on cardiac cells and to determine whether they can modulate the biological activity of 7-ketocholesterol (7KC) involved in the development of cardiovascular diseases. Nanoparticles of iron labeled with Texas Red are introduced in cultures of nonbeating mouse cardiac cells (HL1-NB) with or without 7-ketocholesterol 7KC, and their ability to induce cell death, pro-inflammatory and oxidative effects are analyzed simultaneously. Flow cytometry (FCM), confocal laser scanning microscopy (CLSM), and subsequent factor analysis image processing (FAMIS) are used to characterize the action of iron nanoparticles and to define their cytotoxicity which is evaluated by enhanced permeability to SYTOX Green, and release of lactate deshydrogenase (LDH). Pro-inflammatory effects are estimated by ELISA in order to quantify IL-8 and MCP-1 secretions. Pro-oxidative effects are measured with hydroethydine (HE). Iron Texas Red nanoparticles accumulate at t...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2006
To demonstrate CD36 expression with quantum dots (QDs) 525 and/or 605 on human monocytic U937 cel... more To demonstrate CD36 expression with quantum dots (QDs) 525 and/or 605 on human monocytic U937 cells and atherosclerotic tissue sections by means of flow cytometry (FCM) and/or confocal laser scanning microscopy (CLSM). U937 cells and tissue sections were analyzed by means of FCM and/or CLSM. FCM was performed, using different ultraviolet (UV) and visible (488/532 nm) excitation modes. In the visible mode, fluorescence intensities of QDs, phycoerythrin (PE) and fluorescein isothiocyanate (FITC) were compared. Three-dimensional (3-D) sequences of images were obtained by spectral analysis in a CLSM and analyzed by the factor analysis of medical image sequences (FAMIS) algorithm, providing factor curves and images. Factor images are the result of the FAMIS image processing method, which differentiates emission spectra from 3D sequences of images. In CLSM analysis, preparations are screened in a UV excitation mode to optimize the possibilities of QDs and have the benefit of 4',6-diam...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2009
To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse ... more To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse cardiac cells (HL1-NB) cultured without or with 7-ketocholesterol (7KC) found at an increased level in the plasma of atherosclerotic patients and to simultaneously analyze their cytotoxic, proinflammatory and oxidative properties. Flow cytometry (FCM), confocal laser scanning microscopy and subsequent factor analysis image processing were used to characterize the uptake of nanoparticles and to define their cytotoxicity, evaluated by enhanced permeability to SYTOX Green, release of lactate dehydrogenase (LDH) and morphologic nuclear changes determined with Hoechst 33342. Proinflammatory effects were estimated by enzyme linked immunoassay to quantify IL-8 and MCP-1 secretion. The overproduction of reactive oxygen species (ROS) was determined by FCM with hydroethidine. Whereas the nanoparticles had no cytotoxic or inflammatory effects, they could stimulate ROS production. QDs were not incor...
Annual International Conference of the IEEE Engineering in Medicine and Biology Society, 1996
The aim of the authors' study was to provide an efficient computer automated method for real ... more The aim of the authors' study was to provide an efficient computer automated method for real time determination of the passive properties of voltage clamped cells with the smallest possible stimulation amplitude. The method had to be able to determine a wide range of time constants. The authors applied the correlation technique with a binary noise generator. This technique provides
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2009
To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse ... more To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse cardiac cells (HL1-NB) cultured without or with 7-ketocholesterol (7KC) found at an increased level in the plasma of atherosclerotic patients and to simultaneously analyze their cytotoxic, proinflammatory and oxidative properties. Flow cytometry (FCM), confocal laser scanning microscopy and subsequent factor analysis image processing were used to characterize the uptake of nanoparticles and to define their cytotoxicity, evaluated by enhanced permeability to SYTOX Green, release of lactate dehydrogenase (LDH) and morphologic nuclear changes determined with Hoechst 33342. Proinflammatory effects were estimated by enzyme linked immunoassay to quantify IL-8 and MCP-1 secretion. The overproduction of reactive oxygen species (ROS) was determined by FCM with hydroethidine. Whereas the nanoparticles had no cytotoxic or inflammatory effects, they could stimulate ROS production. QDs were not incor...
(i) We studied the effects of a new cromakalim analogue, SR47063, in guinea-pig ventricular cells... more (i) We studied the effects of a new cromakalim analogue, SR47063, in guinea-pig ventricular cells. The experiments were carried out in whole-cell patch clamp with internal and external solutions supposedly similar to the physiological ones. (ii) SR47063 reversibly activated a time-independent current reversing near the potassium equilibrium potential, and a time-dependent current reversing at a more positive potential. Both currents were blocked by application of glibenclamide. (iii) The time-independent and the time-dependent currents were activating for the same concentration of agonist in every cell, this concentration being very different from cell to cell. (iv) The amplitude of the time-dependent current was shown to depend directly neither on agonist concentration nor on potential, but rather on the amplitude of the current flowing during the prepulse before the test pulse. (v) We conclude that SR47063 is a potent KATP channel opener acting at concentrations lower than one micromolar, and that the time-dependent current is likely due to accumulation and depletion of potassium in restricted areas of the cells.
... In Fig. 2(B), the currents for negative voltage steps appear much larger, and time dependent ... more ... In Fig. 2(B), the currents for negative voltage steps appear much larger, and time dependent as earlier9 Note the total A B: 5ms :L 9 IlnA \ 9 J, OOmV: -_ ~-~_~-- __-----__. - , C ImnA D 40 80 Emv = / ~ -1 ~2-to -2 c o -3 E i-4 ~.1 -4o -~0 EmV Fig. ...
Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2007
The aim of this work was to study, in vitro, cell colonization of two biomaterials currently used... more The aim of this work was to study, in vitro, cell colonization of two biomaterials currently used for bone and cartilage repair, this step being important to understand the function of engineered tissues. Current methods that use histological approaches are not always suited to tissue-engineering analysis. We, therefore, set up a protocol to assess cell distribution, utilizing noninvasive confocal microscopy and fluorescent labels with a far red emission wavelength to optimize scaffold transparency and minimize light scattering. Hard (ceramic substitute) and soft (collagen sponge) biomaterials were seeded respectively, on one side of the scaffold, with human fibroblasts and bovine chondrocytes labelled with carbocyanine dyes (DiD and DiR). The mean penetration depth for DiR labelled fibroblasts and chondrocytes in the two scaffolds, around 270 m, was greater than for DiD (136-218 microm) labelled cells. These depths were independent of cell origin but were influenced by the nature of the scaffolds. Collagen sponge is transparent in contrast to ceramic substitutes where measurements could only be made in opened macropores. Besides the limits of the equipment, the limits of the supports were diffusion for collagen sponges and transmission for ceramic substitutes. Confocal microscopy techniques could thus be used to address the question of cell colonization of porous biomaterials in a noninvasive manner.
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