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 Software-Based Three-Dimensional Deconvolution Microscopy of Cytoskeletal Proteins in Cultured Fibroblast Using Open-Source Software and Open Hardware

2019 ◽  
Vol 5 (12) ◽  
pp. 88
Author(s):  
Kazuo Katoh
Keyword(s):  
Open Source ◽  
Open Source Software ◽  
Laser Scanning ◽  
Low Cost ◽  
Three Dimensional ◽  
Cytoskeletal Proteins ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Actual Measurement ◽  
Open Hardware

As conventional fluorescence microscopy and confocal laser scanning microscopy generally produce images with blurring at the upper and lower planes along the z-axis due to non-focal plane image information, the observation of biological images requires “deconvolution.” Therefore, a microscope system’s individual blur function (point spread function) is determined theoretically or by actual measurement of microbeads and processed mathematically to reduce noise and eliminate blurring as much as possible. Here the author describes the use of open-source software and open hardware design to build a deconvolution microscope at low cost, using readily available software and hardware. The advantage of this method is its cost-effectiveness and ability to construct a microscope system using commercially available optical components and open-source software. Although this system does not utilize expensive equipment, such as confocal and total internal reflection fluorescence microscopes, decent images can be obtained even without previous experience in electronics and optics.

scholarly journals Progress towards Sustainable Control of Xylella fastidiosa subsp. pauca in Olive Groves of Salento (Apulia, Italy)

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 668
Author(s):  
Marco Scortichini ◽  
Stefania Loreti ◽  
Nicoletta Pucci ◽  
Valeria Scala ◽  
Giuseppe Tatulli ◽  
...  
Keyword(s):  
Control Strategy ◽  
Laser Scanning ◽  
Vascular System ◽  
Xylella Fastidiosa ◽  
Low Cost ◽  
Interdisciplinary Studies ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Olive Groves

Xylella fastidiosa subsp. pauca is the causal agent of “olive quick decline syndrome” in Salento (Apulia, Italy). On April 2015, we started interdisciplinary studies to provide a sustainable control strategy for this pathogen that threatens the multi-millennial olive agroecosystem of Salento. Confocal laser scanning microscopy and fluorescence quantification showed that a zinc-copper-citric acid biocomplex—Dentamet®—reached the olive xylem tissue either after the spraying of the canopy or injection into the trunk, demonstrating its effective systemicity. The biocomplex showed in vitro bactericidal activity towards all X. fastidiosa subspecies. A mid-term evaluation of the control strategy performed in some olive groves of Salento indicated that this biocomplex significantly reduced both the symptoms and X. f. subsp. pauca cell concentration within the leaves of the local cultivars Ogliarola salentina and Cellina di Nardò. The treated trees started again to yield. A 1H-NMR metabolomic approach revealed, upon the treatments, a consistent increase in malic acid and γ-aminobutyrate for Ogliarola salentina and Cellina di Nardò trees, respectively. A novel endotherapy technique allowed injection of Dentamet® at low pressure directly into the vascular system of the tree and is currently under study for the promotion of resprouting in severely attacked trees. There are currently more than 700 ha of olive groves in Salento where this strategy is being applied to control X. f. subsp. pauca. These results collectively demonstrate an efficient, simple, low-cost, and environmentally sustainable strategy to control this pathogen in Salento.

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.

Research on Printed Conductive Silver Layer based on Inkjet Photo Paper

2021 ◽  
Author(s):  
Yi Fang ◽  
Lixin Mo ◽  
Zhiqing Xin ◽  
Yinjie Chen ◽  
Xiu Li ◽  
...  
Keyword(s):  
High Performance ◽  
Laser Scanning ◽  
Low Cost ◽  
Flexible Substrate ◽  
Electronic Devices ◽  
Laser Scanning Microscopy ◽  
Silver Layer ◽  
Confocal Laser ◽  
Nano Silver ◽  
Silver Ink

Printed electronics is an emerging technology that applies traditional printing or coating processes to the manufacture of electronic devices and products. In order to find a low-cost, high-performance, environmentally-friendly flexible substrate suitable for electronic devices, the printability between four kinds of inkjet photo papers and nano-silver ink was investigated. First, different surface morphologies of the inkjet photo papers were measured by a confocal laser scanning microscopy. Then, a pen and a gravure printer were used to test the printability between photo papers and nano-silver ink. It was found that the conductive track and pattern was influenced by the surface morphology of the photo papers. Furthermore, a four-probe test showed that the conductivity of the ink layers on the four photo papers was almost at the same level. Furthermore, a tearing test with 3 M tapes showed that the silk photo paper had the best tearing resistance. In general, silk photo paper has the best overall performance. This research could be beneficial for the development of flexible electronic devices which are low-cost, mass manufacture suitable and environment friendly.

scholarly journals Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET)

2003 ◽  
Vol 31 (5) ◽  
pp. 1020-1027 ◽  
Author(s):  
D.S. Lidke ◽  
P. Nagy ◽  
B.G. Barisas ◽  
R. Heintzmann ◽  
J.N. Post ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Laser Scanning ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

We report the implementation and exploitation of fluorescence polarization measurements, in the form of anisotropy fluorescence lifetime imaging microscopy (rFLIM) and energy migration Förster resonance energy transfer (emFRET) modalities, for wide-field, confocal laser-scanning microscopy and flow cytometry of cells. These methods permit the assessment of rotational motion, association and proximity of cellular proteins in vivo. They are particularly applicable to probes generated by fusions of visible fluorescence proteins, as exemplified by studies of the erbB receptor tyrosine kinases involved in growth-factor-mediated signal transduction.

scholarly journals Microscope-Based Imaging Platform for Large-Scale Analysis of Oral Biofilms

2012 ◽  
Vol 78 (24) ◽  
pp. 8703-8711 ◽  
Author(s):  
L. Karygianni ◽  
M. Follo ◽  
E. Hellwig ◽  
D. Burghardt ◽  
M. Wolkewitz ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Large Scale ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Wide Field ◽  
Bacterial Composition ◽  
Entire Area ◽  
Content Type ◽  
Oral Biofilms

ABSTRACTA microscopic method for noninvasively monitoring oral biofilms at the macroscale was developed to describe the spatial distribution of biofilms of different bacterial composition on bovine enamel surfaces (BES). For this purpose, oral biofilm was grownin situon BES that were fixed at approximal sites of individual upper jaw acrylic devices worn by a volunteer for 3 or 5 days. Eubacteria,Streptococcusspp., andFusobacterium nucleatumwere stained using specific fluorescencein situhybridization (FISH) probes. The resulting fluorescence signals were subsequently tested by confocal laser scanning microscopy (CLSM) and monitored by an automated wide-field microscope-based imaging platform (Scan∧R). Automated image processing and data analysis were conducted by microscope-associated software and followed by statistical evaluation of the results. The full segmentation of biofilm images revealed a random distribution of bacteria across the entire area of the enamel surfaces examined. Significant differences in the composition of the microflora were recorded across individual as well as between different enamel surfaces varying from sparsely colonized (47.26%) after 3 days to almost full surface coverage (84.45%) after 5 days. The enamel plates that were positioned at the back or in the middle of the oral cavity were found to be more suitable for the examination of biofilms up to 3 days old. In conclusion, automated microscopy combined with the use of FISH can enable the efficient visualization and meaningful quantification of bacterial composition over the entire sample surface. Due to the possibility of automation, Scan∧R overcomes the technical limitations of conventional CLSM.

scholarly journals Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells

2022 ◽  
Vol 5 (1) ◽  
Author(s):  
Nazar Oleksiievets ◽  
Yelena Sargsyan ◽  
Jan Christoph Thiele ◽  
Nikolaos Mougios ◽  
Shama Sograte-Idrissi ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Laser Scanning ◽  
Super Resolution ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Wide Field ◽  
Fluid Exchange ◽  
Multiplexed Imaging ◽  
Bio Imaging ◽  
A Cell

AbstractDNA point accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful super-resolution technique highly suitable for multi-target (multiplexing) bio-imaging. However, multiplexed imaging of cells is still challenging due to the dense and sticky environment inside a cell. Here, we combine fluorescence lifetime imaging microscopy (FLIM) with DNA-PAINT and use the lifetime information as a multiplexing parameter for targets identification. In contrast to Exchange-PAINT, fluorescence lifetime PAINT (FL-PAINT) can image multiple targets simultaneously and does not require any fluid exchange, thus leaving the sample undisturbed and making the use of flow chambers/microfluidic systems unnecessary. We demonstrate the potential of FL-PAINT by simultaneous imaging of up to three targets in a cell using both wide-field FLIM and 3D time-resolved confocal laser scanning microscopy (CLSM). FL-PAINT can be readily combined with other existing techniques of multiplexed imaging and is therefore a perfect candidate for high-throughput multi-target bio-imaging.

Different patterns of rDNA distribution in Pisum sativum nucleoli correlate with different levels of nucleolar activity

1993 ◽  
Vol 104 (3) ◽  
pp. 843-852 ◽  
Author(s):  
M. I. Highett ◽  
D. J. Rawlins ◽  
P. J. Shaw
Keyword(s):  
Pisum Sativum ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Ccd Camera ◽  
Cell Types ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Wide Field ◽  
Root Tips ◽  
Root Cells

We have used in situ hybridization with probes to rDNA, labelled either with digoxygenin or directly with fluorescein, to determine the arrangement of these genes within the nucleoli of Pisum sativum L. root cells. Confocal laser scanning microscopy was used to image the three-dimensional structures revealed, but we have also compared this technique with deconvolution of conventional (wide-field) fluorescence images measured with a cooled CCD camera, and have shown that the results are remarkably similar. When the deconvolution technique was applied to the confocal data it gave clearer images than could be achieved by confocal microscopy alone. We have analysed the distribution of rDNA in the different cell types observable in root tips: the quiescent centre; active meristematic cells; and relatively differentiated root cap, epidermal and cortical cells. In addition to four perinucleolar knobs of condensed, inactive rDNA genes, corresponding to the four nucleolar organizers in P. sativum, which were the most brightly labelled structures, several characteristic patterns of intranucleolar labelling were apparent, including bright foci, large central chromatin masses, and fine, decondensed interconnecting fibres. The larger and more active the nucleolus, the smaller the proportion of condensed perinucleolar rDNA. In some large and active meristematic nucleoli, all the internal rDNA is decondensed, showing that transcription cannot be restricted to the bright foci, and is most likely to occur on the decondensed fibres.

scholarly journals Confocal Microscopy Predicts the Risk of Recurrence and Malignant Transformation of Mucocutaneous Neurofibromas in NF-1: An Observational Study

2018 ◽  
Vol 2018 ◽  
pp. 1-5
Author(s):  
Giuseppe Giudice ◽  
Giorgio Favia ◽  
Angela Tempesta ◽  
Luisa Limongelli ◽  
Michelangelo Vestita
Keyword(s):  
Laser Scanning ◽  
Low Cost ◽  
Surgical Excision ◽  
Neurofibromatosis Type ◽  
Optical Microscope ◽  
Laser Scanning Microscopy ◽  
Depth Of Field ◽  
Confocal Laser ◽  
Malignant Degeneration ◽  
Risk Of Recurrence

From 2005 to 2010, 20 consecutive patients with fully manifested neurofibromatosis type 1 (NF1) underwent elective neurofibroma resection at our institution (Departments of Plastic Surgery and of Odontostomatology). Specimens were photographed under optical microscope and confocal laser scanning microscopy (CLSM) with ultra-high accuracy of detail, including depth of field. Patients were followed up for a minimum of 4 years and up to a maximum of 12 years, postsurgery. While all nonrecurring lesions showed intense fluorescence, six of the seven lesions with absence of fluorescence under CLSM recurred at a mean of 5.5 years after surgical excision. Among the re-excised lesions, 3 were diagnosed as malignant at the subsequent removal. Despite the limitation of a small cohort, CLSM appears to be a simple and low-cost technique to differentiate forms of neurofibromas with low and high risk of recurrence and malignant degeneration.

scholarly journals Application of confocal laser scanning microscopy to the study of amber bioinclusions

2021 ◽  
Vol 4 (3) ◽  
Author(s):  
YAN-ZHE FU ◽  
YAN-DA LI ◽  
YI-TONG SU ◽  
CHEN-YANG CAI ◽  
DI-YING HUANG
Keyword(s):  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
General Procedure ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Wide Field ◽  
Micro Computed Tomography ◽  
Power Capability

Confocal laser scanning microscopy is an essential analytical tool in biological, biomedical, and material sciences, integrating microscope manufacturing technology, optical-electronic technology, and computer technology. In the last decade, confocal laser scanning microscopy has been successfully applied to the study of amber bioinclusions. Enhanced signal to noise ratios, resolution power, capability of optical sectioning, three-dimensional reconstruction, and better performance when imaging thicker samples provide a great deal of valuable and detailed morphological information about amber fossils. We briefly discuss the practical applications of CLSM in amber studies and compare it with other imaging methods commonly used in the field, including bright-field microscopy, wide-field fluorescence microscopy, and micro-computed tomography. A general procedure for imaging amber inclusions with CLSM is provided, with a focus on pretreatments and image processing.

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