Biomedical subjects

Tim Nielsen

Publications and source records attributed to Tim Nielsen.

4 recordsLinked to original sources

PubMed2005

Two-dimensional imaging without scanning by multifocal multiphoton microscopy.

We describe multifocal multiphoton microscopy giving images without laser scanning. A multitude of 8 x 8 laser beams is focused into a sample yielding two-photon excitation in a plane. The focal spots are arranged in a rectangular array with close spacing between individual points (approximately 0.5 microm). The fluorescence emission from the sample is recorded with a CCD camera, but, owing to the close distance between the beams, they can no longer be regarded as individual points but rather as an illumination of the plane that is covered by the array of focal points. The axial sectioning capability is comparable with an ordinary single-beam two-photon microscope. Interference between the beams that could compromise the axial sectioning capability does not occur in our setup owing to small temporal delays between the individual beams. The axial sectioning capability of the setup is discussed in detail by means of the step response in which the foci are scanned axially into a uniformly fluorescent medium.

Equipment Design↗

PubMed2005

Endocytosis of Xanthomonas campestris pathovar campestris lipopolysaccharides in non-host plant cells of Nicotiana tabacum.

The specific recognition of phytopathogenic bacteria by plant cells is generally mediated by a number of signal molecules. The elicitor-active lipopolysaccharides (LPS) of the phytopathogenic bacterium Xanthomonas campestris pv. campestris (X.c.c) are recognized by its non-host plant Nicotiana tabacum (N.t.). This LPS was purified and labelled with fluorescein isothiocyanate (FITC) for monitoring the fate of these signal molecules in intact plant cells of tobacco. In this study we were able to show that the so-labelled LPS rapidly bound to the cell wall and was then internalized into the cells in a temperature- and energy-dependent way. This uptake of LPS could be outcompeted by the addition of an excess of unlabelled LPS. Furthermore, it was blocked by amantadine, an inhibitor of receptor-mediated endocytosis of mammalian cells. Immunolocalization experiments showed for the first time a significant co-localization of the LPS-elicitor with endosomal structures using an anti-Ara6 antibody. These observations suggest specific endocytosis of LPS(X.c.c.) into tobacco cells. The possibility for a receptor-mediated endocytosis comparable to the mammalian system will be discussed.

Cells, Cultured↗

PubMed2004

In vivo two-photon laser-scanning microscopy of Ca2+ dynamics in visual motion-sensitive neurons.

We applied two-photon laser-scanning microscopy (TPLSM) to motion-sensitive visual interneurons of the fly to study Ca(2+) dynamics in vivo at a higher spatial and temporal resolution than possible with conventional fluorescence microscopy. Based on a custom-built two-photon microscope, we performed line scans to measure changes in presynaptic Ca(2+) concentrations elicited by visual stimulation. We used a fast avalanche photodiode (APD) with a high quantum efficiency to detect even low levels of emitted fluorescence. Our experiments show that our in vivo preparation is amenable to TPLSM: with excitation intensities low enough not to cause photodamage, activity-dependent fluorescence changes of Ca(2+)-sensitive dyes can be detected in small neuronal branches. The performance of two-photon and conventional Ca(2+) imaging carried out consecutively at the same neuron is compared and it is demonstrated that two-photon imaging allows us to detect differences in Ca(2+) dynamics between individual neurites.

Animals↗

PubMed2004

Influence of temperature, modified atmosphere packaging, and heat treatment on aroma compounds in broccoli.

The aroma compounds in broccoli stored in different modified atmospheres were studied. The packaging materials used were oriented polypropylene (OPP), poly(vinyl chloride) (PVC), and low-density polyethylene (LDPE) containing an ethylene-absorbing sachet. All samples were stored for either 1 week at a constant temperature of 10 degrees C or for 3 days at 4 degrees C, followed by 4 days at 10 degrees C. The atmospheres that developed inside the packaging materials differed significantly. The broccoli samples were analyzed raw and after cooking, with regard to volatile compounds, using gas-phase (headspace) extraction followed by gas chromatography-mass spectrometry (GC-MS) and GC-olfactometry. Dimethyl sulfide (DMS), dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS), hexanal, 3-cis-hexen-1-ol, nonanal, ethanol, and a group of thiocyanates were selected for a detailed study because these compounds cause off-odor and can be used as indicators of stress. Significant differences were found in the aroma profiles of the broccoli samples relative to the packaging materials used for storage. Storage in OPP (14% O(2), 10.5% CO(2)) resulted in most of the off-odors, while storage in LDPE (6% O(2), 7% CO(2)) and PVC (17.9% O(2), 4% CO(2)) was found to maintain the concentration of DMS, DMDS, and DMTS during storage. Heat treatment of the broccoli increased the content of aroma compounds as well as the number of compounds containing sulfur.

Brassica↗