Relevant bibliographies by topics / CLSM (confocal laser scanning microscopy)

Academic literature on the topic 'CLSM (confocal laser scanning microscopy)'

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Published: 9 March 2023
Last updated: 11 March 2023

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Journal articles on the topic "CLSM (confocal laser scanning microscopy)"

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Raarup, Merete Krog, and Jens Randel Nyengaard. "QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY." Image Analysis & Stereology 25, no. 3 (May 3, 2011): 111. http://dx.doi.org/10.5566/ias.v25.p111-120.

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This paper discusses recent advances in confocal laser scanning microscopy (CLSM) for imaging of 3D structure as well as quantitative characterization of biomolecular interactions and diffusion behaviour by means of one- and two-photon excitation. The use of CLSM for improved stereological length estimation in thick (up to 0.5 mm) tissue is proposed. The techniques of FRET (Fluorescence Resonance Energy Transfer), FLIM (Fluorescence Lifetime Imaging Microscopy), FCS (Fluorescence Correlation Spectroscopy) and FRAP (Fluorescence Recovery After Photobleaching) are introduced and their applicability for quantitative imaging of biomolecular (co-)localization and trafficking in live cells described. The advantage of two-photon versus one-photon excitation in relation to these techniques is discussed.
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Zucker, Robert M. "Confocal Microscopy System Performance: Axial Resolution." Microscopy Today 12, no. 1 (January 2004): 38–40. http://dx.doi.org/10.1017/s1551929500051816.

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The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. When tests are made to evaluate the performance of a CLSM, the usual subjective assessment is accomplished by using a histological test slide to create a “pretty picture.” Without the use of functional tests many of the machines may be working at sub-optimal performance levels, delivering sub optimum performance, and possibly misleading data. In order to replace the subjectivity in evaluating a confocal microscope, tests were derived or perfected that measure field illumination, lens clarity, laser power, laser stability, dkhroic functionality, spectral registration, axial resolution, scanning stability, PMT quality, overall machine stability, and system noise (1-3). It is anticipated by using this type of test data, performance standards for confocal microscopes will be obtained and the current subjectivity in evaluating CLSM performance will be eliminated.
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Zucker, Robert M. "Confocal Microscopy System Performance: Field Illumination." Microscopy Today 10, no. 5 (September 2002): 8–13. http://dx.doi.org/10.1017/s1551929500058284.

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The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The reliability of the CLSM to obtain specific measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. For many applications it is useful to know the CLSM system's performance prior to acquiring data images so the necessary resolution, sensitivity and precision can be obtained. Applications involving deconvolution, FRET and quantification necessitate that the confocal microscope is correctly configured and operating at the highest performance levels.
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Chladil, Ladislav, Hana Hálová, and Ondřej Čech. "In-situ Confocal Laser Microscopy Study of Lead Sulfate Crystal Growth on Negative Electrode of Lead-acid Batteries." ECS Transactions 105, no. 1 (November 30, 2021): 159–66. http://dx.doi.org/10.1149/10501.0159ecst.

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Confocal Laser Scanning Microscopy (CLSM) is a widely used technique mainly in fields of biology or multidisciplinary material sciences. Although CLSM has the ability to monitor also electrochemical processes like lead sulfate-crystal growth, nobody used CLSM for such application. We performed operando observation of the pasted active mass of negative electrode for lead-acid batteries during deep cycling. Electrode with pasted negative active mass was optimized for cycling in ECC-opto-std electrochemical cell by EL-CELL. Lead sulfate crystal growth and changes of electrode surface during cycling were observed using a laser scanning confocal microscope Olympus Lext OLS4100. We evaluate the surface changes and sulfate crystal growth. The cycling mode leads to fast gradual degradation of the negative electrode and massive growth of lead sulfate crystals. Confocal laser scanning microscopy was identified as a powerful technique for visualization of lead sulfate crystal promotion during battery cycling.
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Balzano, Angela, Klemen Novak, Miha Humar, and Katarina Čufar. "Application of confocal laser scanning microscopy in dendrochronology." Les/Wood 68, no. 2 (December 30, 2019): 5–17. http://dx.doi.org/10.26614/les-wood.2019.v68n02a01.

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We used the Confocal Laser Scanning Microscope (CLSM) Olympus LEXT OLS5000 for non-destructive observation and image analysis of wood anatomy traits in growth layers of tree species from different climatic zones. In European beech (Fagus sylvatica), where tree rings can generally be recognised, we discuss the changes in tree-ring structure due to adverse effects (insect attacks). Growth layers in Mediterranean Aleppo pine (Pinus halepensis) from south-eastern Spain are not always annual and contain numerous intra-annual density fluctuations (IADFs). Ocote pine (Pinus oocarpa) growing at high elevation in Honduras showed growth layers with clear growth ring boundaries and IADFs. In both pines, CLSM allowed us to recognise and measure tracheid parameters to define density fluctuations. In tropical true mahogany (Swietenia macrophylla) from Venezuela and cedrela (Cedrela odorata) from Costa Rica, we studied the growth layers with variable dimensions of vessels demarcated by marginal axial parenchyma.
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Ockleford, Colin. "The confocal laser scanning microscope (CLSM)." Journal of Pathology 176, no. 1 (May 1995): 1–2. http://dx.doi.org/10.1002/path.1711760102.

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Čufar, Katarina, Angela Balzano, Luka Krže, and Maks Merela. "Wood identification using non-destructive confocal laser scanning microscopy." Les/Wood 68, no. 2 (December 30, 2019): 19–29. http://dx.doi.org/10.26614/les-wood.2019.v68n02a02.

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Exact wood identification is usually based on observation of wood features under the microscope. For this, we have to take a sample of the wood from the object and cut thin slides, possibly of all three anatomical sections. Such destructive sampling is often not possible on valuable historical objects, and therefore there is a need for non-destructive approaches. The objective of the study is to present the potential of Confocal Laser Scanning Microscopy (CLSM) using an Olympus LEXT OLS5000 for the identification of wood. We present work on an example of a gothic sculpture, “St. George Defeating the Dragon”. Conventional sampling and microscopical wood identification showed that St. George is made of Norway spruce (Picea abies), and the dragon of poplar (Populus sp.) or willow (Salix sp.). We present crucial features needed for the identification of these species and the limitations with identification if the samples are too small. Finally, we demonstrate the possibility of wood identification of the abovementioned species using CLSM on wood samples without special preparation of the surfaces. CLSM enabled us to observe all the features needed for wood identification.
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Kubanova, A. A., V. V. Chikin, YU YU Shtirshneider, and O. R. Katunina. "Confocal laser scanning microscopy in vivo for diagnosing melanocytic skin neoplasms." Vestnik dermatologii i venerologii 90, no. 3 (June 24, 2014): 85–94. http://dx.doi.org/10.25208/0042-4609-2014-90-3-85-94.

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The authors discuss the use of confocal laser scanning microscopy in vivo (CLSM) for diagnosing melanocytic skin neoplasms and its value for early diagnostics of melanoma. CLSM is an innovation noninvasive visual examination method for real-time multiple and painless examinations of the patient’s skin without injuring the skin integument. The method ensures early diagnostics of skin melanomas with high sensitivity and specificity, which makes it possible to use CLSM for screening melanocytic skin neoplasms for the sake of the early onset of treatment to save patient life and health.
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Pioch, T., S. Stotz, H. J. Staehle, and H. Duschner. "Applications of Confocal Laser Scanning Microscopy to Dental Bonding." Advances in Dental Research 11, no. 4 (November 1997): 453–61. http://dx.doi.org/10.1177/08959374970110041201.

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The introduction of confocal laser scanning microscopy (CLSM) has provided a valuable new technique for the visualization of bonding structures such as a hybrid layer in dentin (Watson, 1989, 1991), In the case of seven commercially-available dentin bonding systems, it could be demonstrated that the CLSM renders considerably more detailed information than the SEM because of its nondestructive nature and because of the possibility of a distinction between components of bonding agents. With most of the bonding systems, measurements of the thickness of the hybrid layer could be carried out when the primer component was labeled with rhodamine B. It was found that this thickness is significantly increased by increases in etching time and only slightly decreased by increases in the drying time of the dentin and of the primer. When rhodamine B was used for dye penetration tests on four different dentin bonding systems, a leakage within the demineralized zone in the dentin was found in each of the specimens. This structure appears similar to that which Sano et al. (1995) called "nanoleakage". The amount of nanoleakage could not be measured by this method. In the case of enamel or ceramic bonding, a penetration zone was found which corresponded to the etching patterns found in enamel and ceramics, respectively. We conclude that CLSM can offer a wealth of new information about bonding morphology and, therefore, should be used in addition to conventional methods so that the maximum information can be obtained.
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Kim, Yoon Soo, and Adya Singh. "Imaging Degraded Wood by Confocal Microscopy." Microscopy Today 6, no. 4 (May 1998): 14–15. http://dx.doi.org/10.1017/s1551929500067225.

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The application of confocal laser scanning microscopy (CLSM) in the studies of biological materials is rapidly expanding because of the opportunity to produce sharp, high resolution images through optical sectioning and computer assisted 3-D reconstruction. At our institute CLSM is being used in a wide range of forestry and wood science studies.Recently we investigated the potential usefulness of CLSM in characterizing biologically degraded wood. The following are images produced from an archaeological wood which has been buried in a wet environment (rice field) for nearly 2,000 years in South Korea and is apparently degraded by bacteria. In an attempt to develop suitable techniques which can be used for routine examination of fragile degraded wood with CLSM, we have compared two different embedding methods for their suitability in preserving the integrity of cells. The embedding media are paraffin wax and LR White resin.
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Dissertations / Theses on the topic "CLSM (confocal laser scanning microscopy)"

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Amin, Anish Kiritkumar. "Chondrocyte death in injured articular cartilage : in vitro evaluation of chondroprotective strategies using confocal laser scanning microscopy." Thesis, University of Edinburgh, 2011. http://hdl.handle.net/1842/5687.

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A reproducible in vitro model of mechanically injured (scalpel cut) articular cartilage was developed in this work utilising bovine and human osteochondral tissue. Using fluorescence-mode confocal laser scanning microscopy (CLSM), the model allowed (1) spatial and temporal quantification of in situ (within the matrix) chondrocyte viability following a full thickness cartilage injury and (2) serial evaluation of three chondroprotective strategies in injured bovine and human articular cartilage: (a) medium osmolarity (b) medium calcium concentration and, (c) subchondral bone attachment to articular cartilage. Medium osmolarity significantly influenced superficial zone chondrocyte death in injured (scalpel cut) bovine and human articular cartilage. Greatest percentage cell death occurred at 0 mOsm (distilled water). Conversely, a raised medium osmolarity (600 mOsm) was chondroprotective. The majority of in situ cell death occurred within 2.5 hours of the experimental injury, with no further increase over 7 days. Exposure of articular cartilage to calcium-free media significantly decreased superficial zone chondrocyte death in injured (scalpel cut) articular cartilage compared with exposure to calcium-rich media (2-20 mM). In calcium-rich media, the extent of percentage cell death increased with increasing medium calcium concentration but remained localised to the superficial zone of injured articular cartilage over 7 days. However, in calcium-free media, there was an increase in percentage cell death within deeper zones of injured articular cartilage over 7 days. Excision of subchondral bone from injured (scalpel cut) articular cartilage resulted in an increase in chondrocyte death at 7 days that occurred in the superficial zone of injured as well as the adjacent uninjured regions of articular cartilage. However, the presence of subchondral bone in the culture medium prevented this increase in chondrocyte death within the superficial zone. Subchondral bone may have interacted with articular cartilage via soluble mediator(s) that influenced chondrocyte survival. In human articular cartilage, healthy subchondral bone also interacted with articular cartilage in explant culture and promoted in situ chondrocyte survival, while sclerotic subchondral bone was detrimental to chondrocyte viability. These findings are of translational relevance to fluid management systems used during open and arthroscopic articular surgery, clinical and experimental research into cartilage injury, repair and degeneration as well as current techniques of tissue engineering.
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Weidhase, Michael, Patrick Beckers, Christoph Bleidorn, and M. Teresa Aguado. "On the role of the proventricle region in reproduction and regeneration in Typosyllis antoni (Annelida: Syllidae)." Universitätsbibliothek Leipzig, 2016. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-216141.

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Background: Syllids are a species rich annelid family possessing remarkable regenerative ability, which is not only the response after traumatic injury, but also a key step during the life cycle of several syllid taxa. In these animals the posterior part of the body becomes an epitoke and is later detached as a distinct unit named stolon. Such a sexual reproductive mode is named schizogamy or stolonization. The prostomium and the proventricle, a modified foregut structure, have been proposed to have a control function during this process, though the concrete mechanisms behind it have never been elucidated. Results: By using different experimental set-ups, histology and immunohistochemistry combined with subsequent cLSM analyzes, we investigate and document the regeneration and stolonization in specimens of Typosyllis antoni that were amputated at different levels throughout the antero-posterior body axis. The removal of the anterior end including the proventricle implies an incomplete anterior regeneration as well as severe deviations from the usual reproductive pattern, i.e. accelerated stolonization, masculinization and the occurrence of aberrant stolons. The detailed anatomy of aberrant stolons is described. A histological study of the proventricle revealed no signs of glandular or secretory structures. The ventricle and the caeca are composed of glandular tissue but they are not involved in the reproductive and regenerative processes. Conclusions: As in other investigated syllids, the proventricle region has a significant role during stolonization and reproduction processes in Typosyllis antoni. When the proventricle region is absent, anterior and posterior regeneration are considerably deviated from the general patterns. However, proventricle ultrastructure does not show any glandular component, thereby questioning a direct involvement of this organ itself in the control of reproduction and regeneration. Our findings offer a comprehensive starting point for further studies of regeneration and reproductive control in syllids as well as annelids in general.
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Haridoss, Sujithera. "In vivo assessment of focal adhesion kinase (FAK) activity in breast cancer cells using fluorescence resonance energy transfer (FRET) sensor and confocal laser scanning microscope (CLSM)." Thesis, Högskolan i Skövde, Institutionen för hälsa och lärande, 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-15706.

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Focal Adhesion Kinase (FAK) is essential for cell migration and plays an important role in tumor metastasis. However, the complex intermolecular and intramolecular interactions that regulate FAK activity at the focal adhesion remain unresolved. We have engineered a toolbox of Fluorescence Resonance Energy Transfer (FRET) sensors for the assessment of FAK activity in human breast cancer cells (MCF-7). Major activity of cancerous cells is drastically growth of the cell in an uncontrollable manner in such cases our human anatomy system normally consists of cell growth activity. The important protein involved in cell functionality in the human body is FAK, due to FAK activity, cell motility, proliferation, survival has been managed in the human body hence, it is necessary to investigate the performance ofFAK activity on breast cancer becomes important. In our study, the differences in bleed through between zoom = 1 and for zoom >1 for donor and acceptor was evaluated. There were no significant differences in Pearson correlation coefficient and bleed through coefficient for both the zooms. With recent advances influorescent probes, instrumentation and methodologies, FRET is sure to revolutionize scientific research in the near future.
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Helm, Conrad, and María Capa. "Comparative analyses of morphological characters in Sphaerodoridae and allies (Annelida) revealed by an integrative microscopical approach." Universitätsbibliothek Leipzig, 2015. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-159898.

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Sphaerodoridae is a group of benthic marine worms (Annelida) characterized by the presence of spherical tubercles covering their whole surface. They are commonly considered as belonging to Phyllodocida although sistergroup relationships are still far from being understood. Primary homology assessments of their morphological features are lacking, hindering the appraisal of evolutionary relationships between taxa. Therefore, our detailed morphological investigation focuses on different Sphaerodoridae as well as on other members of Phyllodocida using an integrative approach combining scanning electron microscopy (SEM) as well as immunohistochemistry with standard neuronal (anti-5-HT) and muscular (phalloidin-rhodamine) markers and subsequent CLSM analysis of whole mounts and sections. Furthermore, we provide histological (HES) and light microscopical data to shed light on the structures and hypothetical function of sphaerodorid key morphological features. We provide fundamental details into the sphaerodorid morphology supporting a Phyllodocida ancestry of these enigmatic worms. However, the muscular arrangement and the presence of an axial muscular pharynx is similar to conditions observed in other members of the Errantia too. Furthermore, nervous system and muscle staining as well as SEM and histological observations of different types of tubercles indicate a homology of the so called microtubercles, present in the long-bodied sphaerodorids, to the dorsal cirri of other Errantia. The macrotubercles seem to represent a sphaerodorid autapomorphy based on our investigations. Therefore, our results allow comparisons concerning morphological patterns between Sphaerodoridae and other Phyllodocida and constitute a starting point for further comparative investigations to reveal the evolution of the remarkable Sphaerodoridae.
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小林, 正典. "共焦点レーザー走査顕微鏡(Confocal Laser Scanning Microscopy : CLSM)による生体関節軟骨と人工軟組織のトライボロジーに関する研究." 京都大学 (Kyoto University), 2003. http://hdl.handle.net/2433/148895.

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Chen, X. "TAGGING BIOCONTROL STREPTOMYCES TO STUDY LETTUCE COLONIZATION." Doctoral thesis, Università degli Studi di Milano, 2015. http://hdl.handle.net/2434/345187.

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The ability of the biological control agents (BCAs) to colonize plant tissues is an important feature involved in microbe-assisted plant protection. Plant-microbe interaction research increased especially in the last decade thanks to technological revolution. Molecular methods and the development of advanced microscopic techniques allow researchers to explore gene expression and localization of beneficial microorganisms within plants. The green fluorescent protein (GFP) and its modified version, enhanced GFP (EGFP), more adapt for expression in mammalian cells and GC-rich actinomycetes like Streptomyces, have been widely used as markers to study gene expression, as well as plant-microbe interactions. Aside fluorescent protein approaches, fluorescence in situ hybridization (FISH) is another frequently used technique to visualize microbial colonization patterns and community composition by application of specific fluorescent probes. Firstly, we transformed five Streptomyces strains, which showed strong inhibition activity against Sclerotinia sclerotiorum, with the EGFP construct by the conjugation method. The conjugation efficiencies varied between the strains, but were comparable to the reference strain. The fitness of transformed strains was similar to wild-type; the transformants maintained similar sporulation, mycelium growth rate, and the ability to produce important secondary metabolites and lytic enzymes. Secondly, two transformed strains, Streptomyces cyaneus ZEA17I, and Streptomyces sp. SW06W, were used to study lettuce colonization dynamics by seed coating method. Their spatio-temporal dynamics were determined in sterile substrate. The strains were consistently recovered from lettuce rhizosphere and inner root tissues up to six weeks. Finally, the colonization pattern of lettuce by Streptomyces cyaneus ZEA17I was examined by both EGFP and FISH approaches combined with confocal laser scanning microscopy (CLSM). For FISH-CLSM analysis, universal bacteria and Streptomyces genus specific probes were used to label S. cyaneus ZEA17I. The consistent presence of the labeled strain at the lettuce root one week after sowing showed that Streptomyces spores could rapidly germinate and produce filamentous mycelium on lettuce. S. cyaneus ZEA17I was detected also on two-week-old roots, indicating the long-term survival ability of this strain in lettuce rhizosphere. Altogether, the antagonistic activity, rhizosphere and root competence showed by the Streptomyces conferred their potential to act as BCA. Further studies on the complex host-pathogen-antagonist interactions will provide additional knowledge to understand the modes and mechanisms of Streptomyces-mediated plant protection.
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Murtin, Chloé Isabelle. "Traitement d’images de microscopie confocale 3D haute résolution du cerveau de la mouche Drosophile." Thesis, Lyon, 2016. http://www.theses.fr/2016LYSEI081/document.

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La profondeur possible d’imagerie en laser-scanning microscopie est limitée non seulement par la distance de travail des lentilles de objectifs mais également par la dégradation de l’image causée par une atténuation et une diffraction de la lumière passant à travers l’échantillon. Afin d’étendre cette limite, il est possible, soit de retourner le spécimen pour enregistrer les images depuis chaque côté, or couper progressivement la partie supérieure de l’échantillon au fur et à mesure de l‘acquisition. Les différentes images prises de l’une de ces manières doivent ensuite être combinées pour générer un volume unique. Cependant, des mouvements de l’échantillon durant les procédures d’acquisition engendrent un décalage non seulement sur en translation selon les axes x, y et z mais également en rotation autour de ces même axes, rendant la fusion entres ces multiples images difficile. Nous avons développé une nouvelle approche appelée 2D-SIFT-in-3D-Space utilisant les SIFT (scale Invariant Feature Transform) pour atteindre un recalage robuste en trois dimensions de deux images. Notre méthode recale les images en corrigeant séparément les translations et rotations sur les trois axes grâce à l’extraction et l’association de caractéristiques stables de leurs coupes transversales bidimensionnelles. Pour évaluer la qualité du recalage, nous avons également développé un simulateur d’images de laser-scanning microscopie qui génère une paire d’images 3D virtuelle dans laquelle le niveau de bruit et les angles de rotations entre les angles de rotation sont contrôlés avec des paramètres connus. Pour une concaténation précise et naturelle de deux images, nous avons également développé un module permettant une compensation progressive de la luminosité et du contraste en fonction de la distance à la surface de l’échantillon. Ces outils ont été utilisés avec succès pour l’obtention d’images tridimensionnelles de haute résolution du cerveau de la mouche Drosophila melanogaster, particulièrement des neurones dopaminergiques, octopaminergiques et de leurs synapses. Ces neurones monoamines sont particulièrement important pour le fonctionnement du cerveau et une étude de leur réseau et connectivité est nécessaire pour comprendre leurs interactions. Si une évolution de leur connectivité au cours du temps n’a pas pu être démontrée via l’analyse de la répartition des sites synaptiques, l’étude suggère cependant que l’inactivation de l’un de ces types de neurones entraine des changements drastiques dans le réseau neuronal
Although laser scanning microscopy is a powerful tool for obtaining thin optical sections, the possible depth of imaging is limited by the working distance of the microscope objective but also by the image degradation caused by the attenuation of both excitation laser beam and the light emitted from the fluorescence-labeled objects. Several workaround techniques have been employed to overcome this problem, such as recording the images from both sides of the sample, or by progressively cutting off the sample surface. The different views must then be combined in a unique volume. However, a straightforward concatenation is often not possible, because the small rotations that occur during the acquisition procedure, not only in translation along x, y and z axes but also in rotation around those axis, making the fusion uneasy. To address this problem we implemented a new algorithm called 2D-SIFT-in-3D-Space using SIFT (scale Invariant Feature Transform) to achieve a robust registration of big image stacks. Our method register the images fixing separately rotations and translations around the three axes using the extraction and matching of stable features in 2D cross-sections. In order to evaluate the registration quality, we created a simulator that generates artificial images that mimic laser scanning image stacks to make a mock pair of image stacks one of which is made from the same stack with the other but is rotated arbitrarily with known angles and filtered with a known noise. For a precise and natural-looking concatenation of the two images, we also developed a module progressively correcting the sample brightness and contrast depending on the sample surface. Those tools we successfully used to generate tridimensional high resolution images of the fly Drosophila melanogaster brain, in particular, its octopaminergic and dopaminergic neurons and their synapses. Those monoamine neurons appear to be determinant in the correct operating of the central nervous system and a precise and systematic analysis of their evolution and interaction is necessary to understand its mechanisms. If an evolution over time could not be highlighted through the pre-synaptic sites analysis, our study suggests however that the inactivation of one of these neuron types triggers drastic changes in the neural network
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Uys, Charlene Ethel. "Preparation and characterisation of pheroid vesicles / Charlene Ethel Uys." Thesis, North-West University, 2006. http://hdl.handle.net/10394/1669.

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Mu, Wangzhong. "Microstructure and Inclusion Characteristics in Steels with Ti-oxide and TiN Additions." Doctoral thesis, KTH, Tillämpad processmetallurgi, 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-162284.

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Non-metallic inclusions in steels are generally considered to be detrimental for mechanical properties. However, it has been recognized that certain inclusions, such as Ti-oxide and TiN, can serve as potent nucleation sites for the formation of intragranular ferrite (IGF) in low-alloy steels. The formation of IGF could improve the toughness of the coarse grained heat affected zone (CGHAZ) of weld metals. Thus, the present thesis mainly focuses on the effect of size of nucleation sites on the IGF formation. Quantitative studies on the composition, size distribution and nucleation probability for each size of the inclusions as well as the area fraction, starting temperature and morphology of an IGF have been carried out. In the present work, the Ti-oxide and TiN powders were mixed with metallic powders. The mixed powders were heated up to the liquid state and cooled with a slow cooling rate of 3.6 ºC/min. These as-cast steels with Ti-oxide and TiN additions were used to simulate the IGF formation in the CGHAZ of weld metals. Specifically, the inclusion and microstructure characteristics in as-cast steels have been investigated. The results show that the nucleant inclusion was identified as a TiOx+MnS phase in steels with Ti2O3 additions and as a TiN+Mn-Al-Si-Ti-O+MnS phase in steels with TiN additions. In addition, the TiOx and TiN phases are detected to be the effective nucleation sites for IGF formation. It is clearly shown that an increased inclusion size leads to an increased probability of IGF nucleation. This probability of IGF nucleation for each inclusion size of the TiOx+MnS inclusions is clearly higher than that of the complex TiN+Mn-Al-Si-Ti-O+MnS inclusions. In addition, the area fraction of IGF in the steels with Ti2O3 additions is larger than that of the steels with TiN additions. This result agrees with the predicted tendency of the probability of IGF nucleation for each inclusion size in the steels with Ti2O3 and TiN additions. In order to predict the effective inclusion size for IGF formation, the critical diameters of the TiO, TiN and VN inclusions, which acted as the nucleation sites of IGF formation, were also calculated based on the classical nucleation theory. The critical diameters of TiO, TiN and VN inclusions for IGF formation were found to be 0.192, 0.355 and 0.810 μm in the present steels. The calculation results were found to be in agreement with the experiment data of an effective inclusion size. Moreover, the effects of the S, Mn and C contents on the critical diameters of inclusions were also calculated. It was found that the critical diameter of the TiO, TiN and VN inclusions increases with an increased content of Mn or C. However, the S content doesn’t have a direct effect on the critical diameter of the inclusions for IGF formation. The probability of IGF nucleation for each inclusion size slightly decreases in the steel containing a higher S content. This fact is due to that an increased amount of MnS precipitation covers the nucleant inclusion surface. In the as-cast experiment, it was noted that an IGF can be formed in steels with Ti2O3 and TiN additions with a cooling rate of 3.6 ºC/min. In order to control the microstructure characteristics, such as the area fraction and the morphology of an IGF, and to investigate the starting temperature of IGF and grain boundary ferrite (GBF) formation, the dynamic transformation behavior of IGF and GBF was studied in-situ by a high temperature confocal laser scanning microscope (CLSM). Furthermore, the chemical compositions of the inclusions and the morphology of IGF after the in-situ observations were investigated by using scanning electron microscopy (SEM), electron backscatter diffraction (EBSD) and electron probe microanalysis (EPMA) which equipped wavelength dispersive spectrometer (WDS). The results show that the area fraction of IGF is larger in the steels with Ti2O3 additions compared to the steels with TiN additions, after the same thermal cycle has been imposed. This is due to that the TiOx phase provides more potent nucleation sites for IGF than the TiN phase does. Also, the area fraction of IGF in the steels is highest after at an intermediate cooling rate of 70 ºC/min, since the competing phase transformations are avoided. This fact has been detected by using a hybrid methodology in combination with CLSM and differential scanning calorimetry (DSC). In addition, it is noted that the morphology of an IGF is refined with an increased cooling rate.

QC 20150325

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Kerschnitzki, Michael. "Bone material characteristics influenced by osteocytes." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2012. http://dx.doi.org/10.18452/16479.

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In dieser Doktorarbeit wird die Hypothese geprüft, ob Osteozyten einen direkten Einfluss auf die Knocheneigenschaften in ihrer unmittelbaren Umgebung haben. Der zentrale Experimentieransatz ist dabei die Korrelation der Organisation des Osteozytennetzwerks mit den Mineraleigenschaften des Knochens auf der Submikrometerebene. Es wird gezeigt, dass bereits die anfängliche Ausrichtung der Osteoblasten entscheidend für die Synthese von hoch ausgerichtetem Knochenmaterial ist. Die dabei entstehenden Osteozytennetzwerke sind so organisiert, dass die Osteozyten und ihre Zellfortsätze jeweils einen möglichst kleinen Abstand zum Knochenmineral haben. Deshalb wird vermutet, dass genau diese Netzwerkorganisation mitentscheidend ist, wie gut die Zellen das Mineral in ih-rer Umgebung beeinflussen können. Messungen der Knochenmineraleigenschaften auf Submikrometerebene mit Röntgenkleinwinkelstreuung bestätigen diese Vermutung. Dabei wird deutlich, dass Knochenmaterial in der Nähe der Osteozyten durch andere Mineraleigenschaften geprägt ist. Um zu klären, wie Osteozyten Mineral in ihrer direkten Umgebung verändern können, werden Mechanismen der passiven Mineralherauslösung aus der mineralisierten Oberfläche des Osteozytennetzwerks untersucht. Es wird gezeigt, dass kalziumarme ionische Lösungen unter physiologischen Bedingungen große Mengen von Kalzium-Ionen aus dem Knochen lösen und diese dann durch die Osteozytennetzwerkstrukturen diffundieren können. Zum Abschluss wurde medullärer Knochen von Hühnern als ein Modellsystem für rasanten Knochenumbau untersucht. Dieser spezielle Knochentyp dient den Hennen als labiles Kalziumreservoir und ermöglicht dadurch die tägliche Eierschalenproduktion. Experimente am medullären Knochen-material zeigen insbesondere die Bedeutung von weniger stabilen Mineralstrukturen die benötigt werden um den Knochen an den schnellen, sich wiederholenden Knochenauf- sowie Abbau optimal anzupassen.
This thesis aims to test the hypothesis whether osteocytes have a direct influence on bone material properties in their vicinity. In this regard, the concomitant ana-lysis of osteocyte network organization and bone ultrastructural properties on the submicron level is the central approach to answer this question. In this work, it is shown that already initial cell-cell alignment during the process of bone formation is crucial for the synthesis of highly organized bone. Furthermore it is proposed that the occurrence of highly ordered osteocyte networks visualized with confocal laser scanning microscopy (CLSM) has a strong impact on the ability of osteocytes to directly influence bone material properties. These highly organized networks are another consequence of initial cell-cell alignment and are found to be arranged such as to feature short mineral cell distances. Examination of sub-micron mineral properties with scanning small angle x-ray scattering (sSAXS) shows that bone material in the direct vicinity of osteocytes and their cell proc-esses shows different mineral properties compared to bone further away in the depth of the tissue. Moreover, mechanisms of passive mineral extraction from the mineralized surface of the osteocyte network, due to the treatment with calcium poor ionic solutions, are investigated. It is shown that this chemical process occurring under physiological conditions leads not only to the dissolution of considerable amounts of calcium, but also to efficient diffusion of these ions through the osteocyte network structures. Finally, medullary bone which is intended as a labile calcium source for daily egg shell formation in hens is used as a model system for rapid bone turnover rates. This bone type in particular indicates the importance of uniquely adapted, less stable mineral structures to fit the requirements for rapid bone resorption as well as reformation.
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Books on the topic "CLSM (confocal laser scanning microscopy)"

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David, Shotton, and Royal Microscopical Society, eds. Confocal laser scanning microscopy. Oxford: BIOS Scientific in association with the Royal Microscopical Society, 1997.

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W, Jones Christopher, ed. Confocal laser scanning microscopy in orthopaedic research. Amsterdam: Elsevier, 2005.

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Division, Bio-Rad Microscopy. MRC-1024: Laser scanning confocal imaging system : user operating manual, Issue 2.0. Hemel Hempstead: Bio-Rad Microscopy Division, 1996.

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Guthoff, Rudolf F., Christophe Baudouin, and Joachim Stave. Atlas of Confocal Laser Scanning In-vivo Microscopy in Ophthalmology. Berlin, Heidelberg: Springer Berlin Heidelberg, 2006. http://dx.doi.org/10.1007/3-540-32707-x.

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Brandt, Roland, and Lidia Bakota. Laser scanning microscopy and quantitative image analysis of neuronal tissue. New York: Humana Press, 2014.

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Guthoff, Rudolf. Atlas of confocal laser scanning in-vivo microscopy in opthalmology [i.e. ophthalmology]: Principles and applications in diagnostic and therapeutic ophtalmology [i.e. ophthalmology]. Berlin: Springer, 2006.

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Sheppard, C. J. R., and David Shotton. Confocal Laser Scanning Microscopy. CRC Press LLC, 2021.

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Sheppard, C. J. R., and David Shotton. Confocal Laser Scanning Microscopy. CRC Press LLC, 2021.

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Sheppard, C. J. R., and David Shotton. Confocal Laser Scanning Microscopy. CRC Press LLC, 2021.

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Sheppard, C., and D. Shotton. Confocal Laser Scanning Microscopy. Springer Singapore Pte. Limited, 1997.

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Book chapters on the topic "CLSM (confocal laser scanning microscopy)"

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Kihm, Kenneth D. "Confocal Laser Scanning Microscopy (CLSM)." In Near-Field Characterization of Micro/Nano-Scaled Fluid Flows, 55–79. Berlin, Heidelberg: Springer Berlin Heidelberg, 2011. http://dx.doi.org/10.1007/978-3-642-20426-5_4.

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Sandison, David R., Rebecca M. Williams, K. Sam Wells, James Strickler, and Watt W. Webb. "Quantitative Fluorescence Confocal Laser Scanning Microscopy (CLSM)." In Handbook of Biological Confocal Microscopy, 39–53. Boston, MA: Springer US, 1995. http://dx.doi.org/10.1007/978-1-4757-5348-6_3.

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Karygianni, Lamprini, Elmar Hellwig, and Ali Al-Ahmad. "Multiplex Fluorescence In Situ Hybridization (M-FISH) and Confocal Laser Scanning Microscopy (CLSM) to Analyze Multispecies Oral Biofilms." In Methods in Molecular Biology, 65–72. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-0467-9_5.

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Stanciu, G., S. G. Stanciu, C. Dan, Konstantinos M. Paraskevopoulos, Xanthippi Chatzistavrou, E. Kontonasaki, and Petros Koidis. "Surface Topography Characterization of Apatite Formation on Bioactive Glass Modified Dental Ceramics Using Confocal Laser Scanning CLSM) and Environmental Scanning Electron Microscopy (ESEM)." In Bioceramics 18, 689–92. Stafa: Trans Tech Publications Ltd., 2006. http://dx.doi.org/10.4028/0-87849-992-x.689.

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Dutta, Bandita, Moupriya Nag, Dibyajit Lahiri, and Rina Rani Ray. "Analysis of Biofilm Matrix by Multiplex Fluorescence In Situ Hybridization (M-FISH) and Confocal Laser Scanning Microscopy (CLSM) During Nosocomial Infections." In Springer Protocols Handbooks, 183–203. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1378-8_8.

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Wannemacher, Reinhold. "Confocal Laser Scanning Microscopy." In Encyclopedia of Nanotechnology, 1–21. Dordrecht: Springer Netherlands, 2015. http://dx.doi.org/10.1007/978-94-007-6178-0_34-2.

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Olivier, Thomas, and Baptiste Moine. "Confocal Laser Scanning Microscopy." In Optics in Instruments, 1–77. Hoboken, NJ USA: John Wiley & Sons, Inc., 2013. http://dx.doi.org/10.1002/9781118574386.ch1.

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Wannemacher, Reinhold. "Confocal Laser Scanning Microscopy." In Encyclopedia of Nanotechnology, 673–91. Dordrecht: Springer Netherlands, 2016. http://dx.doi.org/10.1007/978-94-017-9780-1_34.

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Vergara-Irigaray, Nuria, Michèle Riesen, Gianluca Piazza, Lawrence F. Bronk, Wouter H. P. Driessen, Julianna K. Edwards, Wadih Arap, et al. "Laser Scanning Confocal Microscopy." In Encyclopedia of Nanotechnology, 1192. Dordrecht: Springer Netherlands, 2012. http://dx.doi.org/10.1007/978-90-481-9751-4_100341.

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Peroulis, Dimitrios, Prashant R. Waghmare, Sushanta K. Mitra, Supone Manakasettharn, J. Ashley Taylor, Tom N. Krupenkin, Wenguang Zhu, et al. "Confocal Laser Scanning Microscopy." In Encyclopedia of Nanotechnology, 500–516. Dordrecht: Springer Netherlands, 2012. http://dx.doi.org/10.1007/978-90-481-9751-4_34.

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Conference papers on the topic "CLSM (confocal laser scanning microscopy)"

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Simard-Normandin, M., and R. Rahman. "Confocal Laser Scanning Microscopy (CLSM), A Tool for Counterfeit Detection." In ISTFA 2018. ASM International, 2018. http://dx.doi.org/10.31399/asm.cp.istfa2018p0064.

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Abstract This paper explains the CLSM technique and presents surface roughness measurement data from several groups of known authentic and suspect counterfeit parts. Surface roughness is an important characteristic of plastic encapsulated or metal lidded parts because counterfeit parts are often blacktopped or re-polished and remarked.
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Kinzer, David. "A video rate confocal laser scanning microscope." In OSA Annual Meeting. Washington, D.C.: Optica Publishing Group, 1992. http://dx.doi.org/10.1364/oam.1992.wi1.

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A video rate confocal laser scanning microscope (CLSM) for biological applications has been developed for manufacture. This microscope is used for viewing thin "optical sections" of a sample by rejecting the signal from out-of-focus planes. The microscope consists of a multiline argon ion laser, a galvanometer and acousto-optic deflector (AOD) for video-rate scanning, and a lens system for coupling into a standard optical microscope stand. Performance of better than 1/4 wave peak-to-peak OPD is achieved for visible wavelengths, and operation has been extended to the UV (364 nm) for UV flash photolysis studies. This presentation will describe the design, performance tests, and experiences in manufacture of the CLSM. Techniques for handling chromatic aberration and the wavelength-dependent scan of the AOD will be presented.
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Draaijer, A., and P. M. Houpt. "A Real-Time Confocal Laser Scanning Microscope (CLSM)." In Hague International Symposium, edited by Ludwig J. Balk and Tony Wilson. SPIE, 1987. http://dx.doi.org/10.1117/12.941501.

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Shilling, Meghan, Lipiin Sung, and Thomas R. Kurfess. "Mesoscale Edge Measurement Using a Confocal Laser Scanning Microscope." In ASME 2006 International Manufacturing Science and Engineering Conference. ASMEDC, 2006. http://dx.doi.org/10.1115/msec2006-21096.

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In many mesoscale parts (> 100μm) with microscale features (100nm to 100μm), the edge can constitute a large percentage of the total feature size. These edges need to be measured and characterized. Unfortunately, very few tools exist that are able to measure a range of angles with a resolution appropriate to these parts. The measurement methods commonly used to characterize mesoscale parts are designed for nominally planar surfaces and fail when used on a surface with moderate slope. Other tools exist which can adequately capture a sloping surface, however they have characteristics which make them less desirable for edge measurement (contact, destructive, image-output). The confocal laser scanning microscope (CLSM) is a non-contact method which has special properties that allow it to measure sloping surfaces of various materials and finishes. This paper will discuss the CLSM and its ability to measure edges of mesoscale parts.
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Shang, Hongpeng, DeGui Sun, and Huilin Jiang. "Waveguide Roughness Measuring Metrology with Confocal Laser Scanning Microscope (CLSM)." In 2018 IEEE International Conference on Manipulation, Manufacturing and Measurement on the Nanoscale (3M-NANO). IEEE, 2018. http://dx.doi.org/10.1109/3m-nano.2018.8552185.

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Vennat, E., W. Wang, R. Genthial, B. David, E. Dursun, and A. Gourrier. "Three Dimensional Characterization of the Dentin Porous Network Using Confocal Laser Scanning Microscopy (CLSM)." In Sixth Biot Conference on Poromechanics. Reston, VA: American Society of Civil Engineers, 2017. http://dx.doi.org/10.1061/9780784480779.116.

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Wang, Xin, Patrick Kwon, Ruslan Pelikhatyy, and Dave (Dae-Wook) Kim. "Characterization of Fiber Pull-Outs in Drilled CFRP Holes Using Confocal Laser Microscope." In ASME 2014 International Manufacturing Science and Engineering Conference collocated with the JSME 2014 International Conference on Materials and Processing and the 42nd North American Manufacturing Research Conference. American Society of Mechanical Engineers, 2014. http://dx.doi.org/10.1115/msec2014-4115.

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Carbon Fiber Reinforced Plastic (CFRP) material is often drilled when constructing a large aircraft structure. When drilling CFRP, many defects can be left on the CFRP hole surface. One of the most detrimental surface defects is known as fiber pull-outs, which occur when bundles of fibers are pulled away by fiber-matrix de-bonding and matrix stripping. The objective of this research is to use confocal laser scanning microscope (CLSM) in order to characterize fiber pull-outs occurred during the drilling process of quasi-isotropic CFRP. This new optical characterization method is capable of measuring maximum depth and the distributions of fiber pull-outs. Fiber pull-outs are also qualitatively characterized by SEM and CLSM. It is found that the average depth of fiber pull-outs acquired from CLSM is approximately two times larger than those from the surface profilometer. However, there is a proportional correlation of the data between CLSM and surface profilometer.
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Stanciu, G. A., B. Savu, I. Sandulescu, K. Paraskevopoulos, and P. Koidis. "Study of hydroxyl carbonate apatite formation on bioactive glass coated dental ceramics by confocal laser scanning microscopy (CLSM)." In SPIE Proceedings, edited by Dan C. Dumitras, Maria Dinescu, and Vitally I. Konov. SPIE, 2007. http://dx.doi.org/10.1117/12.730204.

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Ishiguro, Hiroshi, and Takashi Horimizu. "Three-Dimensional Microstructure of Muscle Tissues During Freezing and Thawing." In ASME 1999 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 1999. http://dx.doi.org/10.1115/imece1999-0603.

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Abstract Three-dimensional behavior of ice crystals and cells during the freezing and thawing of biological tissues was investigated microscopically in real time by using a confocal laser scanning microscope (CLSM) and a fluorescent dye, acridine orange (AO). Fresh tender meat (2nd pectoral muscles) of chicken was stained with the AO in physiological saline, and then frozen and thawed in a uniform temperature under two different thermal protocols: a) slow-cooling and rapid-warming and b) rapid-cooling and rapid-warming. The CLSM noninvasively produced tomograms of the tissues to clarify the pattern of freezing, morphology of ice crystals in the tissues, and the interaction between ice crystals and cells.
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Schlu¨ter, Michael, Marko Hoffmann, and Norbert Ra¨biger. "Characterization of Micro Fluidic Devices by Optical Measurements." In ASME 2007 5th International Conference on Nanochannels, Microchannels, and Minichannels. ASMEDC, 2007. http://dx.doi.org/10.1115/icnmm2007-30208.

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Micro fluidic devices are successfully in use for several applications in chemical engineering and biotechnology. Nevertheless, there is still no breakthrough for micro process engineering because of a lack in understanding the mechanisms for local hydrodynamics and mass transfer on micro scales. Micro Particle Image Velocimetry (μ-PIV) combined with Confocal Laser Scanning Microscopy (CLSM) enables the measurement of three-dimensional flow and concentration fields in micro devices for common stationary cases. By quantitative analysis of pressure drops, mixing qualities and residence time distributions an adjustment of micro reactor devices for the demands of chemical and biochemical reactions becomes possible.
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Reports on the topic "CLSM (confocal laser scanning microscopy)"

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Tan, Li, Qiong Liu, Yun Chen, Ya-Qiong Zhao, Jie Zhao, Marie Aimee Dusenge, Yao Feng, et al. Efficacy of sonic activation techniques on tubular dentin sealer penetration：A systematic review and meta-analysis. INPLASY - International Platform of Registered Systematic Review and Meta-analysis Protocols, July 2022. http://dx.doi.org/10.37766/inplasy2022.7.0116.

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Review question / Objective: Is sonic activation techniques more effective than conventional needle irrigation for the tubular dentin sealer penetration. The included study was a randomized controlled trial. Eligibility criteria: A comprehensive search was conducted for all published studies evaluating efficacy of percentage and maximum depth of sealer penetration, following the use of SI and standardized irrigants (NaOCl and EDTA). Because this can hardly be measured clinically, only confocal laser scanning microscopy (CLSM) studies were selected owing to wide use of this methodology for evaluating tubular dentin sealer penetration. The studies using previously filled roots or animal teeth, artificial debris, and plastic blocks, and studies measuring the penetration of tubular dentin sealers in lateral root canals, isthmus, or artificial grooves were excluded to maintain the standardized sample selecting and measuring (Virdee et al. 2018). The search was limited to articles published between January 2000 and June 2022 to ensure conclusions were drawn from contemporary data. There are no language restrictions on filtering articles to ensure the integrity of included data.
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Or, Dani, Shmulik Friedman, and Jeanette Norton. Physical processes affecting microbial habitats and activity in unsaturated agricultural soils. United States Department of Agriculture, October 2002. http://dx.doi.org/10.32747/2002.7587239.bard.

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experimental methods for quantifying effects of water content and other dynamic environmental factors on bacterial growth in partially-saturated soils. Towards this end we reviewed critically the relevant scientific literature and performed theoretical and experimental studies of bacterial growth and activity in modeled, idealized and real unsaturated soils. The natural wetting-drying cycles common to agricultural soils affect water content and liquid organization resulting in fragmentation of aquatic habitats and limit hydraulic connections. Consequently, substrate diffusion pathways to soil microbial communities become limiting and reduce nutrient fluxes, microbial growth, and mobility. Key elements that govern the extent and manifestation of such ubiquitous interactions include characteristics of diffusion pathways and pore space, the timing, duration, and extent of environmental perturbations, the nature of microbiological adjustments (short-term and longterm), and spatial distribution and properties of EPS clusters (microcolonies). Of these key elements we have chosen to focus on a manageable subset namely on modeling microbial growth and coexistence on simple rough surfaces, and experiments on bacterial growth in variably saturated sand samples and columns. Our extensive review paper providing a definitive “snap-shot” of present scientific understanding of microbial behavior in unsaturated soils revealed a lack of modeling tools that are essential for enhanced predictability of microbial processes in soils. We therefore embarked on two pronged approach of development of simple microbial growth models based on diffusion-reaction principles to incorporate key controls for microbial activity in soils such as diffusion coefficients and temporal variations in soil water content (and related substrate diffusion rates), and development of new methodologies in support of experiments on microbial growth in simple and observable porous media under controlled water status conditions. Experimental efforts led to a series of microbial growth experiments in granular media under variable saturation and ambient conditions, and introduction of atomic force microscopy (AFM) and confocal scanning laser microscopy (CSLM) to study cell size, morphology and multi-cell arrangement at a high resolution from growth experiments in various porous media. The modeling efforts elucidated important links between unsaturated conditions and microbial coexistence which is believed to support the unparallel diversity found in soils. We examined the role of spatial and temporal variation in hydration conditions (such as exist in agricultural soils) on local growth rates and on interactions between two competing microbial species. Interestingly, the complexity of soil spaces and aquatic niches are necessary for supporting a rich microbial diversity and the wide array of microbial functions in unsaturated soils. This project supported collaboration between soil physicists and soil microbiologist that is absolutely essential for making progress in both disciplines. It provided a few basic tools (models, parameterization) for guiding future experiments and for gathering key information necessary for prediction of biological processes in agricultural soils. The project sparked a series of ongoing studies (at DTU and EPFL and in the ARO) into effects of soil hydration dynamics on microbial survival strategy under short term and prolonged desiccation (important for general scientific and agricultural applications).
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Droby, Samir, Michael Wisniewski, Ron Porat, and Dumitru Macarisin. Role of Reactive Oxygen Species (ROS) in Tritrophic Interactions in Postharvest Biocontrol Systems. United States Department of Agriculture, December 2012. http://dx.doi.org/10.32747/2012.7594390.bard.

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To elucidate the role of ROS in the tri-trophic interactions in postharvest biocontrol systems a detailed molecular and biochemical investigation was undertaken. The application of the yeast biocontrol agent Metschnikowia fructicola, microarray analysis was performed on grapefruit surface wounds using an Affymetrix Citrus GeneChip. the data indicated that 1007 putative unigenes showed significant expression changes following wounding and yeast application relative to wounded controls. The expression of the genes encoding Respiratory burst oxidase (Rbo), mitogen-activated protein kinase (MAPK) and mitogen-activated protein kinase kinase (MAPKK), G-proteins, chitinase (CHI), phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) and 4-coumarate-CoA ligase (4CL). In contrast, three genes, peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT), were down-regulated in grapefruit peel tissue treated with yeast cells. The yeast antagonists, Metschnikowia fructicola (strain 277) and Candida oleophila (strain 182) generate relatively high levels of super oxide anion (O2−) following its interaction with wounded fruit surface. Using laser scanning confocal microscopy we observed that the application of M. fructicola and C. oleophila into citrus and apple fruit wounds correlated with an increase in H2O2 accumulation in host tissue. The present data, together with our earlier discovery of the importance of H₂O₂ production in the defense response of citrus flavedo to postharvest pathogens, indicate that the yeast-induced oxidative response in fruit exocarp may be associated with the ability of specific yeast species to serve as biocontrol agents for the management of postharvest diseases. Effect of ROS on yeast cells was also studied. Pretreatment of the yeast, Candida oleophila, with 5 mM H₂O₂ for 30 min (sublethal) increased yeast tolerance to subsequent lethal levels of oxidative stress (50 mM H₂O₂), high temperature (40 °C), and low pH (pH 4). Suppression subtractive hybridization analysis was used to identify genes expressed in yeast in response to sublethal oxidative stress. Transcript levels were confirmed using semi quantitative reverse transcription-PCR. Seven antioxidant genes were up regulated. Pretreatment of the yeast antagonist Candida oleophila with glycine betaine (GB) increases oxidative stress tolerance in the microenvironment of apple wounds. ROS production is greater when yeast antagonists used as biocontrol agents are applied in the wounds. Compared to untreated control yeast cells, GB-treated cells recovered from the oxidative stress environment of apple wounds exhibited less accumulation of ROS and lower levels of oxidative damage to cellular proteins and lipids. Additionally, GB-treated yeast exhibited greater biocontrol activity against Penicillium expansum and Botrytis cinerea, and faster growth in wounds of apple fruits compared to untreated yeast. The expression of major antioxidant genes, including peroxisomal catalase, peroxiredoxin TSA1, and glutathione peroxidase was elevated in the yeast by GB treatment. A mild heat shock (HS) pretreatment (30 min at 40 1C) improved the tolerance of M. fructicola to subsequent high temperature (45 1C, 20–30 min) and oxidative stress (0.4 mol-¹) hydrogen peroxide, 20–60 min). HS-treated yeast cells showed less accumulation of reactive oxygen species (ROS) than non-treated cells in response to both stresses. Additionally, HS-treated yeast exhibited significantly greater (P≥0.0001) biocontrol activity against Penicillium expansum and a significantly faster (Po0.0001) growth rate in wounds of apple fruits stored at 25 1C compared with the performance of untreated yeast cells. Transcription of a trehalose-6-phosphate synthase gene (TPS1) was up regulated in response to HS and trehalose content also increased.
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Pesis, Edna, Elizabeth J. Mitcham, Susan E. Ebeler, and Amnon Lers. Application of Pre-storage Short Anaerobiosis to Alleviate Superficial Scald and Bitter Pit in Granny Smith Apples. United States Department of Agriculture, January 2013. http://dx.doi.org/10.32747/2013.7593394.bard.

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There is increased demand for high quality fruit produced and marketed with reduced chemical inputs to minimize toxic effects on human health and the environment. Granny Smith (GS) apple quality is reduced by two major physiological disorders, superficial scald and bitter pit (BP). These disorders cause great loss to apple growers worldwide. Superficial scald is commonly controlled by chemical treatments, mainly the antioxidant diphenylamine (DPA) and/or the ethylene action inhibitor, 1-methylcyclopropene (1–MCP). Both chemicals are ineffective in controlling bitter pit incidence. We proposed to investigate the beneficial use of non-chemical, abiotic stress with low O2 (LO2) applied for 10d at 20°C on GS apple fruit. During the project we expanded the treatment to more apple cultivars, Golden Delicious (GD) and Starking Delicious (SD) and another pome fruit, the pear. Apple and pear have similar physiological disorders that develop during cold storage and we examined if the LO2 treatment would also be effective on pear. Application of 0.5% LO2 atmosphere for 10d at 20°C or 500ppb 1-MCP at 20°C prior to cold storage at 0°C, was effective in reducing superficial scald in GS apple. Moreover, LO2 pretreatment was also effective in reducing bitter pit (BP) development in California GS and Israeli GD and SD apples The BP symptoms in GS from California were much more prominent, so the effect of LO2 was more dramatic than the effect on the Israeli cvs. GD and SD, nevertheless the LO2 treatment showed the same trend in all cultivars in reducing BP. The LO2 and 1-MCP -treated fruit exhibited lower levels of ethylene, - farnesene and its oxidation product, 6-methyl-5-hepten-2-one (MHO), as determined by SPME/GC-MS analysis. In addition, LO2 pretreatment applied to California Bartlett or Israeli Spadona pears was effective in reducing superficial scald, senescent scald and internal breakdown after 4 m of cold storage at 0°C. For GS apple, low-temperature storage resulted in oxidative stress and chilling injury, caused by increased production of superoxide anions which in turn led to the generation of other dangerous reactive oxygen species (ROS). Using confocal laser-scanning microscopy and H2O2 measurements of apple peel, we observed ROS accumulation in control fruit, while negligible amounts were found in LO2 and 1-MCP treated fruit. Gene-expression levels of ROS-scavenging enzymes were induced by the various pretreatments: catalase was induced by LO2 treatment, whereas Mn superoxide dismutase was induced by 1-MCP treatment. We assume that LO2 and 1-MCP pretreated fruit remained healthier due to reduced production of ethylene and reactive oxygen substances, such as MHO, during cold storage. The LO2-treated apple exhibited greener peel and firmer fruit after 6 m of cold storage, and the fruit had high crispiness leading to high taste preference. In both pear cultivars, the LO2 treatment led to a reduction in internal breakdown and browning around the seed cavity. We tested the LO2 pre-storage treatment on a semi-commercial scale that would be applicable to a small organic grower by sealing the fruit within the plastic field bins. The treatment was most effective with a continuous flow of nitrogen through the bins; however, a single 6 hour flush of nitrogen was also fairly effective. In addition, we determined that it was very important to have the oxygen levels below 0.5% for approximately 10 days to achieve good scald control, not counting the time required to reduce the oxygen concentration. Our LO2 technology has been proven in this project to be effective in reducing several physiological disorders developed in pome fruit during cold storage. We hope that our non-chemical treatment which is friendly to the environment will be used in the near future for the organic apple and pear industry. The next step should be an analysis of the cost-benefits and commercial feasibility.
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