Developing correlative Optical-Photoacustic microscopy for high-resolution in vivo imaging

PhotoAcoustic Imaging (PAI) is a branch in clinical and pre-clinical imaging, that refers to the techniques mapping acoustic signals caused by the absorption of the short laser pulse. This conversion of electromagnetic energy of the light to the mechanical (acoustic) energy is usually called photoacoustic effect. PAI, by combining optical excitation with acoustical detection, is able to preserve the diffraction limited spatial resolution. At the same time, the penetration depth is extended beyond the diffusive limit. The Laser-Scanning PhotoAcoustic Microscope system (LS-PAM) has been developed, that offers the axial resolution of 7.75 µm with the lateral resolution better than 10 µm. The first in vivo imaging experiments were carried out. Thus, in vivo label-free imaging of the mouse ear was performed. The principle possibility to image vessels located in deep layers of the mouse skin was shown. As well as that, a gold printing sample, vasculature of the Chick Chorioallantoic Membrane Assay, Drosophila larvae were imaged by PAI. During the experimental work, a totally new application of PAM was found, in which the acoustic waves, generated by incident light can be used for further imaging of another sample. In order to enhance the performance of the presented system two main recommendation can be offered. First, the current system should be transformed into reflection-mode setup system. Second, a more powerful source of light with the sufficient repetition rate should be introduced into the system.

Development of 3D super-resolution tomographic STED microscopy and its application to studies on bone resorption

In this doctoral thesis, a tomographic STED microscopy technique for 3D super-resolution imaging was developed and utilized to observebone remodeling processes. To improve upon existing methods, wehave used a tomographic approach using a commercially available stimulated emission depletion (STED) microscope. A certain region of interest (ROI) was observed at two oblique angles: one at a standard inverted configuration from below (bottom view) and another from the side (side view) via a micro-mirror positioned close to the ROI. The two viewing angles were reconstructed into a final tomogram. The technique, named as tomographic STED microscopy, was able to achieve an axial resolution of approximately 70 nm on microtubule structures in a fixed biological specimen. High resolution imaging of osteoclasts (OCs) that are actively resorbing bone was achieved by creating an optically transparent coating on a microscope coverglass that imitates a fractured bone surface. 2D super-resolution STED microscopy on the bone layer showed approximately 60 nm of lateral resolution on a resorption associated organelle allowing these structures to be imaged with super-resolution microscopy for the first time. The developed tomographic STED microscopy technique was further applied to study resorption mechanisms of OCs cultured on the bone coating. The technique revealed actin cytoskeleton with specific structures, comet-tails, some of which were facing upwards and some others were facing downwards. This, in our opinion, indicated that during bone resorption, an involvement of the actin cytoskeleton in vesicular exocytosis and endocytosis is present. The application of tomographic STED microscopy in bone biology demonstrated that 3D super-resolution techniques can provide new insights into biological 3D nano-structures that are beyond the diffraction-limit when the optical constraints of super-resolution imaging are carefully taken into account.

Bioimage informatics in STED super-resolution microscopy

Optical microscopy is living its renaissance. The diffraction limit, although still physically true, plays a minor role in the achievable resolution in far-field fluorescence microscopy. Super-resolution techniques enable fluorescence microscopy at nearly molecular resolution. Modern (super-resolution) microscopy methods rely strongly on software. Software tools are needed all the way from data acquisition, data storage, image reconstruction, restoration and alignment, to quantitative image analysis and image visualization. These tools play a key role in all aspects of microscopy today – and their importance in the coming years is certainly going to increase, when microscopy little-by-little transitions from single cells into more complex and even living model systems. In this thesis, a series of bioimage informatics software tools are introduced for STED super-resolution microscopy. Tomographic reconstruction software, coupled with a novel image acquisition method STED< is shown to enable axial (3D) super-resolution imaging in a standard 2D-STED microscope. Software tools are introduced for STED super-resolution correlative imaging with transmission electron microscopes or atomic force microscopes. A novel method for automatically ranking image quality within microscope image datasets is introduced, and it is utilized to for example select the best images in a STED microscope image dataset.

La pure vérité cachée

[Mazarinade. 1649]

OEuvres de « VINCENT COSSARD », savoir.

Contient : 1 « Livre des causes ou principes et origines des deux Natures » ; 2 « Discours et resolution de la divination, ou Divine et souveraine verité touchant ce qui est à advenir de la guerre des chrestiens et Ismaëlites ou Turchz. De la victoire, en quel lieu sera, et pourquoy. 1575 » ; 3 « La dixieme partie ou section des OEuvres de JAMBLICUS, des Misteres des Egyptiens. 1576 » ; 4 « Consideration du christianisme » ; 5 « La Rome babylonique, ou Petit traicté de la confusion et ruine babylonique en la cité de Rome » ; 6 « Premiere partie de la Demonstration du vespre du monde. 1577 ». Précédée d'une lettre « à Richard Crosnier, presbtre » ; 7 « La Dispute du pape Sylvestre avec les Juifz, celebrée à Rome l'an trois cens apres l'Incarnation, en la presence de l'empereur Constantin et Helene, sa mere, Augustes, touchant la verité de Jesus Christ, Dieu incarné, juxte les antiennes promesses et propheties dont est tres doctement par le meme texte de l'Escripture saincte refutée l'erreur non seulement des Juifz et Mahometains touchant la Trinité, mais aussi de plusieurs heretiques antiens et modernes » ; 8 « De la Naissance, progres et fin de l'empire romain, oeuvre composé, depuis trois cens ans, par ENGELBERT, abbé d'Aumont, faict françois. 1575 » ; 9 « Les quatre Livres de PHILIPPE THEOPH. PARACELSE. De la longue Vie, ou Conservation et prorogation de la vie, faict françoys du latin, l'an. commençant 1576 » ; 10 « Oratio in Assumptione B. Virginis, habita Rothomagi, anno 1576 » ; 11 « Ad senatum urbis novae et fortissimae Portus Gratiae, totius Galliae propugnaculi » ; 12 « In S. Elizabethae, virg. Sconaugiensis, sermonem sextum, sive cap. 4, lib. 2 Viarum Dei sive visionum, inscriptum de Via praelatorum, praefatio ad... Jo. Ruffum, ecclesiae quae est in sylvula Anneburgii antistitem » ; 13 « OΥINKENTIOΣ KOΣΣΑΡΔΟΣ. Ιωάννη τω̃ αυτου̃ πατρι ευ πράττειν », lettre, en grec, de Cossard à son père, 15 décembre 1570 ; 14 Lettres latines, de « GUILLAUME POSTEL à Cossard » ; 3 janvier 1576 (fol. 192); — de « V. COSSARD à G. Postel », 27 septembre 1576 (ibid.), — de « LOUIS MARTEL à V. Cossard », 11 septembre 1576 (fol. 193) ; — de « V. COSSARD à G. Postel », 1er juillet 1577 (ibid.) ; — de « G. POSTEL à V. Cossard », 10 juillet 1577 (fol. 194) ; — de « G. POSTEL à V. Cossard », 2 mai (ibid) ; 15 « Epistre presentée par COSSARD à Mgr le reverendissime archevesque de Raims et abbé de Fescamp, 1576 » ; 16 « Preface à tres affectionné zelateur de la vraye pieté et syncerité du christianisme, Me Robert Joly, prestre, sur la generale et terrible vision faicte à noble et religieuse personne F. Robert, premierement prestre seculier, et depuis, de l'ordre Saint Dominique ; mise en françoys, pour la conversion du monde babylonique, apres avoir été cachée l'espace de 288 ans » ; 17 « Nemo, sive Sermo de vita et rebus gestis viri ad hunc diem incogniti Neminis, videlicet contemporanei Deo Patri, Filio coessentialis, et origine conformis Spiritui Sancto » ; 18 « La Voix du desert » ; 19 « Specilegium exercitationum poeticarum », en vers français ; 20 « Chant royal sur la pure et tres saincte Conception de la Vierge » ; 21 « Ballade sur la mesme Immaculée Conception » ; 22 « Consideration ou elegie sur la mort de Me Romain Cossard, premier aux esleus pour le roy à Rouen » ; 23 « Dixain sur la mort du mesme » ; 24 « Ode sur la mort du dict sieur Esleu, mise à l'entour du cierge bruslant sur le tombeau » ; 25 « Elegie sur la mort de damoyselle Marie Le Bouteiller » ; 26 Lettre à « damoysele Marie Le Bouteiller, mere de la deffuncte, sur la mort de son filz, qui mourust un an apres » ; 27 « Au Roy de la febve », vers ; 28 « Chant royal sur l'Immaculée et saincte Conception » ; 29 « Priere à la Vierge pour le roy » ; 30 « Ballade sur la mesme Conception » ; 31 « Autre ballade » ; 32 Epigrammes traduites d'« ARCHIAS », de « MELEAGRE », de « PALLADE », etc ; 33 « Sonnet sur la tres saincte Conception » ; 34 « Sur le tombeau d'un paovre », traduit « du latin de PONTAN » ; 35 « Quatrain de FRANÇOIS DUFRESNE » ; 36 « Ex Th. Moro », quatrain ; 37 « Horatiana rapsodia pro tumulo », centon de vers d'Horace ; 38 « Tombeau », vers

Using classroom meetings to transfer conflict resolution skills /

This study examined whether daily classroom meetings resulted in the positive transfer of conflict resolution information and skills beyond the formal classroom setting and into the classroom. A control group of sixteen Grade five students received three weeks of conflict resolution training and an experimental group of nineteen Grade five students fi-om the same school received three weeks of conflict resolution training followed by three additional weeks of class meetings. Pretest measures were taken via a scaled questionnaire and short answer questions before the conflict resolution lessons began for the following skills: knowledge of conflict resolution; conflict resolution behaviour; and attitude about using conflict resolution to resolve problems with other people. Posttest measures examined conflict resolution skills following involvement in the study. Students chosen randomly and both teachers were interviewed following the study. The teachers were again interviewed three months after the study. Teacher journal notes rounded out the data. The results of the study indicated that the Grade five boys who participated in three weeks of conflict resolution training did not increase their conflict resolution skills in any of the areas examined. Girls who participated in three weeks of conflict resolution training did not improve in two areas (i.e., behaviour, knowledge) and became less positive about using verbal mediation to resolve conflicts. The Grade five students who participated in three weeks of training and three weeks of class meetings obtained different results. The boys improved significantly in their ability to use verbal mediation to resolve conflicts and were more positive about verbal mediation. They did not become more knowledgeable about verbal mediation. The girls who participated in three weeks of training and three weeks of class meetings were more knowledgeable of conflict resolution and used conflict resolution to solve problems with other people. However, they were significantly less positive about using these skills to resolve problems.

Letter from the Secretary of War, transmitting documents containing the information required by the resolution of Mr. Wright, on the 22d inst. relative to the execution of the act fixing the military peace establishment. --

14th Congress, 1st session, 1815-1816. House. Document no. 33.

Massachusetts militia claims : letter from the Secretary of War, transmitting, in pursuance of a resolution of the House of Representatives of the 15th Dec., 1826, a report upon the subject of the claims of the State of Masachusetts for certain services rendered during the late war.--

20th Congress, 2nd session. House Doc. no.3.

Message from the President of the United States, transmitting a report of the Secretary of State, in obedience to a resolution of the thirteenth inst. "requesting the President to lay before this House such documents relative to the Russian mediation as in his opinion it may not be improper to communicate." --

United States,13th Congress, 2d session, 1813-1814. House. Doc. no. 35.

Mr. Giles' speech, delivered in the Senate of the United States, on Thursday, 24th November, 1808, on the resolution of Mr. Hillhouse, to repeal the embargo laws. --

Caption title.

Mr. Hillhouses' speech, on the resolution to repeal the embargo, November 29, 1808. --

Caption title.

Mr. Pickering's speech in the Senate of the United States, on the resolution offered by Mr. Hillhouse to repeal the several acts laying an embargo, November 30, 1808. --

Caption title.