Search results for "Confocal"

A method for measuring mitochondrial mass and activity

2007

Mitochondria, responsible for the energy-generating process essential for the cell metabolism, differ for the number, localization and activity in animal cells and tissues in relation to the energetic needs. Using fluorescent probes specific for mitochondria, Mitotracker Green (MTG) and Orange (MTO), and Confocal Laser-Scanning Microscope (CLSM), we elaborated a method to measure in vivo the mitochondrial mass and activity, in sea urchin Paracentrotus lividus eggs and embryos. The analysis of captured images, revealed a variation of mitochondrial distribution and an increase of activity after fertilization.

Imaging and force transduction in correlative scanning force and confocal fluorescence microscopy

2018

Correlative scanning force and confocal fluorescence microscopy has been used to study individual molecules, nanoparticles and nanoparticle oligomers. By applying a compressive force via the AFM cantilever, spectral blue and red shifts in the range of several meV/GPa have been observed for single dye molecules and semiconductor quantum dots. Moreover, individual Au nanoparticle dimers linked by a chlorophyll binding protein have been imaged in both modes and plasmonic fluorescence enhancement of the chlorophyll emission of up to a factor of 15 has been found.

Dynamics and interactions of parvoviral NS1 protein in the nucleus

2007

Summary Nuclear positioning and dynamic interactions of viral proteins with nuclear substructures play essen- tial roles during infection with DNA viruses. Visual- ization of the intranuclear interactions and motility of the parvovirus replication protein (NS1) in living cells gives insight into specific parvovirus protein- cellular structure interactions. Confocal analysis of highly synchronized infected Norden Laboratory Feline Kidney cells showed accumulation of nuclear NS1 in discrete interchromosomal foci. NS1 fused with enhanced yellow fluorescence protein (NS1- EYFP) provided a marker in live cells for dynamics of NS1 traced by photobleaching techniques. Fluo- rescence Recovery after…

The future of transmission electron microscopy (TEM) in biology and medicine.

2000

1996

The uses of atomic force microscopy, scanning tunneling microscopy, electron spectroscopic imaging, electron energy loss spectroscopy and low voltage, high resolution scanning electron microscopy in polymer research are reviewed

Survival of Erwinia amylovora in mature apple fruit calyces through the viable but nonculturable (VBNC) state

2009

Aims:  Survival of Erwinia amylovora, causal agent of fire blight in pome fruits and other rosaceous plants, was monitored inside mature apples calyces under some storage conditions utilized in fruit. Methods and Results:  Apple fruit calyces inoculated with two E. amylovora strains and their respective GFP-marked strains were maintained at 26° and 5°C, and the effect of copper treatment was assayed at 0·01 and 0·1 mmol l−1 CuSO4. In nontreated apples at 26°C, part of the population of E. amylovora survived in the ‘viable but nonculturable’ (VBNC) state, whereas at 5°C the majority of the population retained culturability. In copper-treated apples, the whole population adopted the VBNC stat…

The differentiation antigen NY-BR-1 is a potential target for antibody-based therapies in breast cancer

2007

Antibody-based cancer immunotherapy relies on the identification and characterization of target antigens and the development of potent antibodies recognizing the target. Here we report the expression analysis and molecular characterization of the differentiation antigen NY-BR-1, which we previously identified by using the SEREX (serological analysis of recombinant cDNA expression libraries) method. Corroborating methodologies, including mRNA quantitation and immunoblotting show that NY-BR-1 is strongly expressed in >70% of 129 breast tumors. Application of a NY-BR-1 specific antibody demonstrated NY-BR-1 expression in primary and metastastic breast cancers. In contrast, most of the breast c…

Glutathione is recruited into the nucleus in early phases of cell proliferation.

2007

We have studied the possible correlation between nuclear glutathione distribution and the progression of the cell cycle. The former was studied by confocal microscopy using 5-chloromethyl fluorescein diacetate and the latter by flow cytometry and protein expression of Id2 and p107. In proliferating cells, when 41% of them were in the S+G(2)/M phase of the cell cycle GSH was located mainly in the nucleus. When cells reached confluence (G(0)/G(1)) GSH was localized in the cytoplasm with a perinuclear distribution. The nucleus/cytoplasm fluorescence ratio for GSH reached a maximal mean value of 4.2 +/- 0.8 at 6 h after cell plating. A ratio higher than 2 was maintained during exponential cell …

Permeability changes of integrin-containing multivesicular structures triggered by picornavirus entry.

2014

Cellular uptake of clustered α2β1-integrin induces the formation of membrane compartments that subsequently mature into a multivesicular body (MVB). Enhanced internalization mediated by clustered integrins was observed upon infection by the picornavirus echovirus 1 (EVI). We elucidated the structural features of virus-induced MVBs (vMVBs) in comparison to antibody-induced control MVBs (mock infection) by means of high-pressure cryo fixation of cells followed by immuno electron tomography during early entry of the virus. Three-dimensional tomograms revealed a marked increase in the size and complexity of these vMVBs and the intraluminal vesicles (ILVs) at 2 and 3.5 hours post infection (p.i.…

Baculovirus capsid display: a novel tool for transduction imaging

2003

Baculoviruses are enveloped insect viruses that can carry large quantities of foreign DNA in their genome. Baculoviruses have proved to be very promising gene therapy vectors but little is known about their transduction mechanisms in mammalian cells. We show in this study that Autographa californica multiple nuclear polyhedrosis virus capsid is compatible with the incorporation of desired proteins in large quantities. Fusions can be made to the N-terminus or C-terminus of the major capsid protein vp39 without compromising the viral titer or functionality. As an example of the baculovirus capsid display we show a tracking of the baculovirus transduction in mammalian cells by an enhanced gree…