scholarly journals High Resolution 3-D Fluorescence Microscopy: A Comparison of Confocal Laser Scanning Microscopy and a wide-Field Deconvolution Technique

1997 ◽  
Vol 5 (6) ◽  
pp. 10-13 ◽  
Author(s):  
Keyword(s):  
Biological Sciences ◽  
Fluorescence Microscopy ◽  
Light Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Specific Structure ◽  
Confocal Laser ◽  
Wide Field ◽  
Deconvolution Technique ◽  
Transmitted Light Microscopy

Advances in the biological sciences have given rise for the need to visualize microscopic structures of interest. As the requirement to see specific structure and sub-resolution details arise, investigators have turned to fluorescent probes to label and observe these details, which are often undetectable using conventional methods in light microscopy. Fluorescence microscopy offers many advantages for visualizing specific structure and sub-resolution details, which are often undetectable using transmitted light microscopy. Recent advances in fluorescent probe and microscope design, as well as imaging instrumentation designed for fluorescence applications, are now permitting life-science researchers to view details in regions of interest with increasing precision, accuracy, and resolution.

Cuticular structures in the copulatory apparatus of Planorbis planorbis (Linnaeus, 1758) (Gastropoda: Pulmonata: Planorbidae)

2009 ◽  
Vol 18 (1) ◽  
pp. 11-16
Author(s):  
E.V. Soldatenko ◽  
A.A. Petrov
Keyword(s):  
Light Microscopy ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Taxonomic Status ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Copulatory Apparatus ◽  
The Family ◽  
Cuticular Structures

The morphology of the copulatory apparatus and associated cuticular structures in Planorbis planorbis was studied by light microscopy, SEM, TEM and confocal laser scanning microscopy. The significance of these cuticular structures for the taxonomic status of the species and for the systematics of the family Planorbidae in general is discussed.

scholarly journals Morphological and morphometric study of pre-ovigerous and post-ovigerous adults of Tanaisia (Paratanaisia) bragai (Santos, 1934) (Digenea, Eucotylidae)

2017 ◽  
Vol 18 (2) ◽  
Author(s):  
Sthefane D'ávila ◽  
Pedro Paulo de Abreu Manso ◽  
Elizabeth Cristina de Almeida Bessa ◽  
Maria de Lurdes de Azevedo Rodrigues
Keyword(s):  
Light Microscopy ◽  
Confocal Laser Scanning Microscopy ◽  
Body Length ◽  
Laser Scanning ◽  
The Body ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Morphometric Study

The aim of this study was to obtain data on the morphology and morphometry of pre-ovigerous and post-ovigerous adults of the species Tanaisia (Paratanaisia) bragai, using confocal laser scanning microscopy to obtain tomographic images of the suckers and tegument. For morphometric analysis, 45 specimens (30 pre-ovigerous adults and 15 post-ovigerous adults) were measured with the aid of an ocular micrometer coupled to the objective of a photonic microscope. Pre-ovigerous and post-ovigerous adult individuals, stained with Mair carmalumen and mounted in permanent preparations, were analyzed by means of confocal laser scanning microscopy. Positive correlation was detected between the body length and ovary length of post-ovigerous adults (rs: 0.774; p<0.01), as well as between the body length and testes (rs: 0.604 and 0.659; p< 0.05), the body length and the length of uterus (rs: 0.839; p< 0,01) and between the ovary width and egg length (rs: 0.777; p<0.01). Morphological study of the pre-ovigerous adults demonstrated that the ovary and testes develop simultaneously before the development of the uterus and vitelline glands. The acetabulum was detected in pre-ovigerous adults stained with hematoxilin and observed using light microscopy. In these specimens, the acetabulum measured 36.7 ± 6.9 µm (25-50 µm) in width and 39.91 ± 6.8 µm (25-55 µm) in length. The acetabulum was not detected in post-ovigerous adults observed with light microscopy. However, this structure was detected using confocal miscrocopy. In the post-ovigerous specimens, the acetabulum presented a reduced size compared to the pre-ovigerous adults. This may imply that this structure has more functional significance in the larval and pre-ovigerous stages.

Light Microscopy, Confocal Laser Scanning Microscopy And Scanning And Transmission Electron Microscopy Of Cerebellar Golgi Cells.

1999 ◽  
Vol 5 (S2) ◽  
pp. 1302-1303
Author(s):  
O. Castejόn ◽  
P Sims
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Synaptic Connections ◽  
Teleost Fishes ◽  
Golgi Cells ◽  
Transmission Electron ◽  
Molecular Layers

The cerebellar cortex of albino mice, hamsters, teleost fishes, primates and human have been examined by correlative microscopy to study the Golgi cell soma, dendritic processes, axonal plexus and synaptic connections in the granular and molecular layers. For light microscopy (LM) toluidinc blue stained-plastic embedded scmithin sections and Golgi light microscopy preparations were used. For confocal laser scanning microscopy (CLSM) of hamster cerebellum the FM4-64 fluorescent stain was used as intracellular tracer (1). Conventional and high resolution scanning electron microscopy (SEM) of teleost fishes, primates and human were coated with gold-palladium and chromium (2). I Transmission electron microscopy (TEM). either by ullrathin sections or frccze-clching replicas, were examined to characterize synaptic connections in the granular and molecular layers. The Golgi cells appeared in the granular layer as polygonal, stellate, round or fusiform microncurons. 10-25 μm in maximal dimension, surrounded by the granule cell groups. Golgi light microscopy.

Interspecies Uptake of Polymeric Microspheres

1998 ◽  
Vol 550 ◽  
Author(s):  
Chris Thanos ◽  
Maryellen Sandor ◽  
Yong Jong ◽  
Jules Jacob ◽  
Kay-Pong Yip ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Laser Scanning ◽  
Polarized Light ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Optical Sectioning ◽  
Intestinal Tissue ◽  
Polymeric Microspheres ◽  
Particle Uptake

AbstractParticle uptake into intestinal tissue has seen increasing attention due to its implications in drug delivery. We attempted to observe a delivery system in vivo and examine uptake in different species. Microspheres were fabricated from polymers including polyanhydrides and delivered to an isolated loop of intestine in several species. The microspheres contained a dye either conjugated to a protein or incorporated freely and were used to qualitatively detect and locate the spheres in the villi of the length of the small intestine. Microspheres were dispersed, sized by a Coulter particle size analyzer, and characterized by confocal and cross-polarized light microscopy, FTIR and SEM. Coulter analysis revealed microspheres to be generally less than 5 microns in diameter. SEM typically showed homogeneous morphology among groups of microspheres. In vivo uptake experiments were performed in rodents, pigs, and ruminants using various microsphere formulations. Microspheres were delivered into the proximal end of the jejunum of anesthetized animals and allowed adequate transit time to be taken up. Animals were euthanized at various time points for explantation of tissue and sampling of blood. Excised samples were embedded inq polyvinyl alcohol, frozen, and cut into sections ranging between 7 and 14 μm in thickness. Our method of incorporating dyes allowed for simultaneous visualization by visible light microscopy and confocal laser scanning microscopy. Two-fluorochrome fluorescence of the microspheres and optical sectioning confirmed the presence of microspheres within intestinal tissue. The amount of uptake depended on the animal model, the duration of the experiment, and the composition of the microsphere. An assay for either the fluorescent dye, the protein attached to it, or the polymer encapsulating it may enable us to determine intracellular concentrations of mierospheres for the quantification of uptake.

scholarly journals A Selective Ratiometric Fluorescent Probe for No-Wash Detection of PVC Microplastic

Polymers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1588
Author(s):  
Valeria Caponetti ◽  
Alexandra Mavridi-Printezi ◽  
Matteo Cingolani ◽  
Enrico Rampazzo ◽  
Damiano Genovese ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Direct Detection ◽  
Low Cost ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Wide Field ◽  
Fluorescence Intensity Ratio ◽  
Green Fluorescence ◽  
One Pot ◽  
Emerging Pollutant

Microplastics (MP) are micrometric plastic particles present in drinking water, food and the environment that constitute an emerging pollutant and pose a menace to human health. Novel methods for the fast detection of these new contaminants are needed. Fluorescence-based detection exploits the use of specific probes to label the MP particles. This method can be environmentally friendly, low-cost, easily scalable but also very sensitive and specific. Here, we present the synthesis and application of a new probe based on perylene-diimide (PDI), which can be prepared in a few minutes by a one-pot reaction using a conventional microwave oven and can be used for the direct detection of MP in water without any further treatment of the sample. The green fluorescence is strongly quenched in water at neutral pH because of the formation dimers. The ability of the probe to label MP was tested for polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), polystyrene (PS), poly methyl methacrylate (PMMA) and polytetrafluoroethylene (PTFE). The probe showed considerable selectivity to PVC MP, which presented an intense red emission after staining. Interestingly, the fluorescence of the MP after labeling could be detected, under excitation with a blue diode, with a conventional CMOS color camera. Good selectivity was achieved analyzing the red to green fluorescence intensity ratio. UV–Vis absorption, steady-state and time-resolved fluorescence spectroscopy, fluorescence anisotropy, fluorescence wide-field and confocal laser scanning microscopy allowed elucidating the mechanism of the staining in detail.

scholarly journals Direct fluorescence imaging of lignocellulosic and suberized cell walls in roots and stems

AoB Plants ◽  
2020 ◽  
Vol 12 (4) ◽  
Author(s):  
Peter Kitin ◽  
Satoshi Nakaba ◽  
Christopher G Hunt ◽  
Sierin Lim ◽  
Ryo Funada
Keyword(s):  
Cell Walls ◽  
Laser Scanning ◽  
Cell Types ◽  
Plant Evolution ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Chemical Information ◽  
Wide Field ◽  
Tissue Preparation ◽  
Plant Structure

Abstract Investigating plant structure is fundamental in botanical science and provides crucial knowledge for the theories of plant evolution, ecophysiology and for the biotechnological practices. Modern plant anatomy often targets the formation, localization and characterization of cellulosic, lignified or suberized cell walls. While classical methods developed in the 1960s are still popular, recent innovations in tissue preparation, fluorescence staining and microscopy equipment offer advantages to the traditional practices for investigation of the complex lignocellulosic walls. Our goal is to enhance the productivity and quality of microscopy work by focusing on quick and cost-effective preparation of thick sections or plant specimen surfaces and efficient use of direct fluorescent stains. We discuss popular histochemical microscopy techniques for visualization of cell walls, such as autofluorescence or staining with calcofluor, Congo red (CR), fluorol yellow (FY) and safranin, and provide detailed descriptions of our own approaches and protocols. Autofluorescence of lignin in combination with CR and FY staining can clearly differentiate between lignified, suberized and unlignified cell walls in root and stem tissues. Glycerol can serve as an effective clearing medium as well as the carrier of FY for staining of suberin and lipids allowing for observation of thick histological preparations. Three-dimensional (3D) imaging of all cell types together with chemical information by wide-field fluorescence or confocal laser scanning microscopy (CLSM) was achieved.

The megalopal stage of the hydrothermal vent crab Austinograea rodriguezensis Tsuchida & Hashimoto, 2002 (Decapoda: Bythograeidae): a morphological description based on CLSM images

Zootaxa ◽  
2021 ◽  
Vol 5040 (3) ◽  
pp. 365-387
Author(s):  
OLGA DEMIDOW ◽  
TERUE C. KIHARA ◽  
PEDRO MARTÍNEZ ARBIZU ◽  
PAUL F. CLARK
Keyword(s):  
Light Microscopy ◽  
Hydrothermal Vent ◽  
Hydrothermal Vents ◽  
Laser Scanning ◽  
Dorsal Surface ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Morphological Description ◽  
Brachyuran Crab ◽  
Federal Institute

The Bythograeidae is unique amongst brachyuran crab taxa as it is the only family where all 6 genera and 16 species are endemic to hydrothermal vents. During the research conducted by German Federal Institute for Geosciences and Natural Resources to identify inactive polymetallic sulphide deposits along Central and Southeast Indian Ridges, the INDEX project collected from hydrothermal vent fields 6 Bythograeidae megalopae. Entire specimens and dissected appendages were stained, mounted on slides and examined using Light Microscopy and Confocal Laser Scanning Microscopy. Additional molecular analysis using mtCOI confirmed the identification of the specimens as Austinograea rodriguezensis Tsuchida & Hashimoto, 2002. The megalopal stage of A. rodriguezensis shows similarities and distinctions between the characters of two other bythograeid megalopae, Bythograea thermydron Williams, 1980, and B. microps Saint Laurent, 1984. Unlike other brachyuran megalopae, B. thermydron and A. rodriguezensis lack long serrate setae and stout serrate spines on the dactylus of the fifth pereiopod. In both species no coxal spines on the pereiopods were observed. The elliptical carapace of B. thermydron is broader than long vs longitudinally rectangular in A. rodriguezensis. This carapace shape resembles B. microps more than B. thermydron, however, the dorsal surface of B. microps carapace is densely covered in short setae vs only covered with short setae on the anterior 1/4 and the posterior 1/6 length of carapace in A. rodriguezensis. Furthermore, the amount of plumose natatory setae on the pleopods in B. microps is in total larger and more variable, than in A. rodriguezensis. Bythograeid megalopae seem quite generalized and miss specific features that reveal them distinctively as endemic vent crab. A distinction from other species is possible by observing the unique combinations of certain characters. This is the first megalopal stage description of Austinograea and the fourth within the Bythograeidae.  

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