scholarly journals Distribution of Phytoplasmas in Infected Plants as Revealed by Real-Time PCR and Bioimaging

2004 ◽  
Vol 17 (11) ◽  
pp. 1175-1184 ◽  
Author(s):  
Nynne Meyn Christensen ◽  
Mogens Nicolaisen ◽  
Michael Hansen ◽  
Alexander Schulz
Keyword(s):  
Real Time ◽  
Laser Scanning ◽  
Infected Plant ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Euphorbia Pulcherrima ◽  
Pcr Assay ◽  
Sieve Tubes ◽  
Polymerase Chain

Phytoplasmas are cell wall-less bacteria inhabiting the phloem and utilizing it for their spread. Infected plants often show changes in growth pattern and a reduced crop yield. A quantitative real-time polymerase chain reaction (Q-PCR) assay and a bioimaging method were developed to quantify and localize phytoplasmas in situ. According to the Q-PCR assay, phytoplasmas accumulated disproportionately in source leaves of Euphorbia pulcherrima and, to a lesser extent, in petioles of source leaves and in stems. However, phytoplasma accumulation was small or nondetectable in sink organs (roots and sink leaves). For bioimaging, infected plant tissue was stained with vital fluorescence dyes and examined using confocal laser scanning microscopy. With a DNA-sensitive dye, the pathogens were detected exclusively in the phloem, where they formed dense masses in sieve tubes of Catharanthus roseus. Sieve tubes were identified by counterstaining with aniline blue for callose and multiphoton excitation. With a potentiometric dye, not all DNA-positive material was stained, suggesting that the dye stained metabolically active phytoplasmas only. Some highly infected sieve tubes contained phytoplasmas that were either inactive or dead upon staining.

Intravital Real-Time Confocal Laser Scanning Microscopy for the In Situ Evaluation of Nanocarriers

2013 ◽  
pp. 607-620 ◽  
Author(s):  
Yu Matsumoto ◽  
Takahiro Nomoto ◽  
Kazuko Toh ◽  
Horacio Cabral ◽  
Mami Murakami ◽  
...  
Keyword(s):  
Real Time ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser

3-D imaging of cells using a confocal laser scanning microscope and digital image processing

1988 ◽  
Vol 46 ◽  
pp. 96-97
Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Laser Scanning Microscope ◽  
Biochemical Processes ◽  
Adjacent Structures

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.

scholarly journals A sample cell for the study of enzyme-induced carbonate precipitation at the grain-scale and its implications for biocementation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jennifer Zehner ◽  
Anja Røyne ◽  
Pawel Sikorski
Keyword(s):  
Real Time ◽  
Laser Scanning Microscopy ◽  
In Situ Monitoring ◽  
Confocal Laser ◽  
Precipitation Process ◽  
Successful Implementation ◽  
Detailed Knowledge ◽  
Carbonate Precipitation ◽  
Sample Cell

AbstractBiocementation is commonly based on microbial-induced carbonate precipitation (MICP) or enzyme-induced carbonate precipitation (EICP), where biomineralization of $$\text {CaCO}_{3}$$ CaCO 3 in a granular medium is used to produce a sustainable, consolidated porous material. The successful implementation of biocementation in large-scale applications requires detailed knowledge about the micro-scale processes of $$\text {CaCO}_{3}$$ CaCO 3 precipitation and grain consolidation. For this purpose, we present a microscopy sample cell that enables real time and in situ observations of the precipitation of $$\text {CaCO}_{3}$$ CaCO 3 in the presence of sand grains and calcite seeds. In this study, the sample cell is used in combination with confocal laser scanning microscopy (CLSM) which allows the monitoring in situ of local pH during the reaction. The sample cell can be disassembled at the end of the experiment, so that the precipitated crystals can be characterized with Raman microspectroscopy and scanning electron microscopy (SEM) without disturbing the sample. The combination of the real time and in situ monitoring of the precipitation process with the possibility to characterize the precipitated crystals without further sample processing, offers a powerful tool for knowledge-based improvements of biocementation.

scholarly journals Influence of Aging on Corrosion Behaviour of the 6061 Cast Aluminium Alloy

Materials ◽  
2021 ◽  
Vol 14 (8) ◽  
pp. 1821
Author(s):  
Ting He ◽  
Wei Shi ◽  
Song Xiang ◽  
Chaowen Huang ◽  
Ronald G. Ballinger
Keyword(s):  
Aluminium Alloy ◽  
Laser Scanning ◽  
Corrosion Behaviour ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Kelvin Probe ◽  
Force Microscopy ◽  
The Matrix ◽  
6061 Aluminium Alloy

The influence of AlFeSi and Mg2Si phases on corrosion behaviour of the cast 6061 aluminium alloy was investigated. Scanning Kelvin probe force microscopy (SKPFM), electron probe microanalysis (EPMA), and in situ observations by confocal laser scanning microscopy (CLSM) were used. It was found that Mg2Si phases were anodic relative to the matrix and dissolved preferentially without significantly affecting corrosion propagation. The AlFeSi phases’ influence on 6061 aluminium alloy local corrosion was greater than that of the Mg2Si phases. The corroded region width reached five times that of the AlFeSi phase, and the accelerating effect was terminated as the AlFeSi dissolved.

scholarly journals In Situ 3D-Imaging of the Inner Ear Synapses with a Cochlear Implant

Life ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 301
Author(s):  
Kathrin Malfeld ◽  
Nina Armbrecht ◽  
Holger A. Volk ◽  
Thomas Lenarz ◽  
Verena Scheper
Keyword(s):  
Cochlear Implant ◽  
Inner Ear ◽  
Laser Scanning ◽  
3D Imaging ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Sensory Cells ◽  
Residual Hearing ◽  
Preparation Methods

In recent years sensorineural hearing loss was found to affect not exclusively, nor at first, the sensory cells of the inner ear. The sensory cells’ synapses and subsequent neurites are initially damaged. Auditory synaptopathies also play an important role in cochlear implant (CI) care, as they can lead to a loss of physiological hearing in patients with residual hearing. These auditory synaptopathies and in general the cascades of hearing pathologies have been in the focus of research in recent years with the aim to develop more targeted and individually tailored therapeutics. In the current study, a method to examine implanted inner ears of guinea pigs was developed to examine the synapse level. For this purpose, the cochlea is made transparent and scanned with the implant in situ using confocal laser scanning microscopy. Three different preparation methods were compared to enable both an overview image of the cochlea for assessing the CI position and images of the synapses on the same specimen. The best results were achieved by dissection of the bony capsule of the cochlea.

scholarly journals In Situ Characterization ofNitrospira-Like Nitrite-Oxidizing Bacteria Active in Wastewater Treatment Plants

2001 ◽  
Vol 67 (11) ◽  
pp. 5273-5284 ◽  
Author(s):  
Holger Daims ◽  
Jeppe L. Nielsen ◽  
Per H. Nielsen ◽  
Karl-Heinz Schleifer ◽  
Michael Wagner
Keyword(s):  
Wastewater Treatment ◽  
16S Rrna ◽  
Laser Scanning ◽  
Wastewater Treatment Plants ◽  
Carbon Sources ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Rrna Gene ◽  
Phylogenetic Affiliation

ABSTRACT Uncultivated Nitrospira-like bacteria in different biofilm and activated-sludge samples were investigated by cultivation-independent molecular approaches. Initially, the phylogenetic affiliation of Nitrospira-like bacteria in a nitrifying biofilm was determined by 16S rRNA gene sequence analysis. Subsequently, a phylogenetic consensus tree of theNitrospira phylum including all publicly available sequences was constructed. This analysis revealed that the genusNitrospira consists of at least four distinct sublineages. Based on these data, two 16S rRNA-directed oligonucleotide probes specific for the phylum and genus Nitrospira, respectively, were developed and evaluated for suitability for fluorescence in situ hybridization (FISH). The probes were used to investigate the in situ architecture of cell aggregates ofNitrospira-like nitrite oxidizers in wastewater treatment plants by FISH, confocal laser scanning microscopy, and computer-aided three-dimensional visualization. Cavities and a network of cell-free channels inside the Nitrospiramicrocolonies were detected that were water permeable, as demonstrated by fluorescein staining. The uptake of different carbon sources byNitrospira-like bacteria within their natural habitat under different incubation conditions was studied by combined FISH and microautoradiography. Under aerobic conditions, theNitrospira-like bacteria in bioreactor samples took up inorganic carbon (as HCO3 − or as CO2) and pyruvate but not acetate, butyrate, and propionate, suggesting that these bacteria can grow mixotrophically in the presence of pyruvate. In contrast, no uptake by theNitrospira-like bacteria of any of the carbon sources tested was observed under anoxic or anaerobic conditions.

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